Wide hybridization is an effective approach for enhancing the resistance of bread wheat (Triticum aestivum L.) to biotic and abiotic stresses by introducing favorable alien genes (Sepsi et al., 2008). Wheatgrass, ...Wide hybridization is an effective approach for enhancing the resistance of bread wheat (Triticum aestivum L.) to biotic and abiotic stresses by introducing favorable alien genes (Sepsi et al., 2008). Wheatgrass, Thinopyrum intermedium (Host) Barkworth & D.R. Dewey or Agropyron intermedium (Host) Beauvoir (2n = 42; genome formula JJjSjSstst), is a perennial species in the tribe Triticeae and an important source of wheat improvement for biotic and abiotic stress resistance and quality-related traits, such as high grain protein concentration (Chen et al., 1998; 2001; 2003; Han et al., 2004; Li and Wang, 2009). In addition, the ready crossing ability of wheatgrass with various Triticum species has made it popular in germ- plasm development.展开更多
BACKGROUND A series of long non-coding RNAs(lncRNAs)have been reported to play a crucial role in cancer biology.Some previous studies report that lncRNA CDKN2B-AS1 is involved in some human malignancies.However,its ro...BACKGROUND A series of long non-coding RNAs(lncRNAs)have been reported to play a crucial role in cancer biology.Some previous studies report that lncRNA CDKN2B-AS1 is involved in some human malignancies.However,its role in hepatocellular carcinoma(HCC)has not been fully deciphered.AIM To decipher the role of CDKN2B-AS1 in the progression of HCC.METHODS CDKN2B-AS1 expression in HCC was detected by quantitative real-time polymerase chain reaction.The malignant phenotypes of Li-7 and SNU-182 cells were detected by the CCK-8 method,EdU method,and flow cytometry,respectively.RNA immunoprecipitation was executed to confirm the interaction between CDKN2B-AS1 and E2F transcription factor 1(E2F1).Luciferase reporter assay and chromatin immunoprecipitation were performed to verify the binding of E2F1 to the promoter of G protein subunit alpha Z(GNAZ).E2F1 and GNAZ were detected by western blot in HCC cells.RESULTS In HCC tissues,CDKN2B-AS1 was upregulated.Depletion of CDKN2B-AS1 inhibited the proliferation of HCC cells,and the depletion of CDKN2B-AS1 also induced cell cycle arrest and apoptosis.CDKN2B-AS1 could interact with E2F1.Depletion of CDKN2B-AS1 inhibited the binding of E2F1 to the GNAZ promoter region.Overexpression of E2F1 reversed the biological effects of depletion of CDKN2B-AS1 on the malignant behaviors of HCC cells.CONCLUSION CDKN2B-AS1 recruits E2F1 to facilitate GNAZ transcription to promote HCC progression.展开更多
Objective To explore the association between the three polymorphisms [ C825T, C1429T and G(-350)A] of the gene encoding the G protein beta 3 subunit (GNB3) and hypertension by performing a case-control study in th...Objective To explore the association between the three polymorphisms [ C825T, C1429T and G(-350)A] of the gene encoding the G protein beta 3 subunit (GNB3) and hypertension by performing a case-control study in the northern Han Chinese population. Methods We recnaited 731 hypertensive patients and 673 control subjects (the calculated power value was 〉 0.8). Genotyping was performed to identify C825T, C1429T and G(-350)A polymorphisms using the TaqMan assay. Comparisons of allelic and genotypic frequencies between cases and controls were made by using the chi-square test. Logistic regression analyses were performed to investigate the relationships between the three polymorphisms of GNB3 gene under different genetic models (additive, dominant and recessive models). Results The genotype dis- tribution and allele frequencies of C825T, C1429T and G(-350)A polymorphisms did not differ significantly between hypertensive patients and control subjects, either when the full sample was assessed, or when the sample was stratified by gender. No significant association was observed between C825T, C 1429T and G(-350)A polymorphisms and the risk of essential hypertension in any genetic model. Linkage dis- equilibrium was only detected between C825T and C 1429T polymorphisms. Haplotype analyses observed that none of the three estimated haplotypes significantly increased the risk of hypertension. Conclusions Our study suggested that the GNB3 gene polymorphisms [C825T, C 1429T and G(-350)A] were not significantly associated with essential hypertension in northern Han Chinese population.展开更多
The cDNA encoding the G protein αq subunit was isolated from the antennae of Helicoverpa assulta (Guen6e) by reverse transcription polymerase chain reaction (RT-PCR) and named as HassGαq. Sequencing analysis sho...The cDNA encoding the G protein αq subunit was isolated from the antennae of Helicoverpa assulta (Guen6e) by reverse transcription polymerase chain reaction (RT-PCR) and named as HassGαq. Sequencing analysis showed that the fulllength of HassGαq open reading frame (ORF) is 1 062 bp, 353 amino acid residues are encoded. The predicted molecular weights (MW) and isoelectric point (PI) are 41.5 kD and 5.15, respectively. HassGαq gene was then constructed into expression vector pGEX-4T-2 for over expression in prokaryotic cells. The SDS-PAGE and Western blot analysis showed that induced by Isopropyl-β-D-Thiogalactoside (IPTG), the GST-HassGαq fusion protein is expressed in Escherichia coil BL21, and its MW was found to be about 66 kD nearly equal to the predicted. In addition, RT-PCR analysis showed that the expressions of HassGαq are not tissue specific.展开更多
Benzene is an established leukotoxin and leukemogen in humans. We have previously re- ported that exposure of workers to benzene and to benzene metabolite hydroquinone in cultured cells induced DNA-dependent protein k...Benzene is an established leukotoxin and leukemogen in humans. We have previously re- ported that exposure of workers to benzene and to benzene metabolite hydroquinone in cultured cells induced DNA-dependent protein kinase catalytic subunit (DNA-PKcs) to mediate the cellular response to DNA double strand break (DSB) caused by DNA-damaging metabolites. In this study, we used a new, small molecule, a selective inhibitor of DNA-PKcs, 2-(morpholin-4-yl)-benzo[h]chomen-4-one (NU7026), as a probe to analyze the molecular events and pathways in hydroquinone-induced DNA DSB repair and apoptosis. Inhibition of DNA-PKcs by NU7026 markedly potentiated the apoptotic and growth inhibitory effects of hydroquinone in proerythroid leukemic K562 cells in a dose-dependent manner. Treatment with NU7026 did not alter the production of reactive oxygen species and oxidative stress by hydroquinone but repressed the protein level of DNA-PKcs and blocked the induction of the kinase mRNA and protein expression by hydroquinone. Moreover, hydroquinone increased the phos- phorylation of Akt to activate Akt, whereas co-treatment with NU7026 prevented the activation of Akt by hydroquinone. Lastly, hydroquinone and NU7026 exhibited synergistic effects on promoting apop- tosis by increasing the protein levels of pro-apoptotic proteins Bax and caspase-3 but decreasing the protein expression of anti-apoptotic protein Bcl-2. Taken together, the findings reveal a central role of DNA-PKcs in hydroquinone-induced hematotoxicity in which it coordinates DNA DSB repair, cell cycle progression, and apoptosis to regulate the response to hydroquinone-induced DNA damage.展开更多
BACKGROUND Protein phosphatase 2 regulatory subunit B''alpha(PPP2R3A)gene has been reported in other tumors,but the influence of PPP2R3A gene expression on the occurrence,development,and prognosis of hepatocel...BACKGROUND Protein phosphatase 2 regulatory subunit B''alpha(PPP2R3A)gene has been reported in other tumors,but the influence of PPP2R3A gene expression on the occurrence,development,and prognosis of hepatocellular carcinoma(HCC)remains unclear.AIM To investigate whether the PPP2R3A gene could be used to predict tumor recurrence and survival of HCC patients after liver transplantation(LT).METHODS Diseased liver tissues of HCC patients after LT were collected as well as their clinical data and follow-up information.The immunohistochemical method was used to detect the expression of PPP2R3A protein in the tissues of 108 patients with primary liver cancer.Theχ2 test was used to analyze the relationship between PPP2R3A protein expression levels and the clinicopathological features of tumors.The Kaplan-Meier method was used to analyze overall postoperative survival.The COX proportional hazard model was used to analyze adverse prognostic factors.RESULTS Immunohistochemistry showed that the PPP2R3A protein was mainly expressed in the cytoplasm of HCC cells.Compared to corresponding peritumoral tissues,expression was higher in HCC tissues(P≤0.001).Correlation analysis showed that high PPP2R3A expression was correlated with preoperative serum alphafetoprotein(AFP)levels(P=0.003),tumor-node-metastasis-t stage(P≤0.001),and envelope invasion(P=0.001).Univariate analysis showed that overall survival(P≤0.001)and recurrence-free survival(P=0.025)of patients with high PPP2R3A expression(≥4 points)were poor compared to those with low expression(<4 points).The overall survival rates or recurrence-free survival rates at 1,2,and 3 years with high PPP2R3A expression were 73%,38%,and 23%or 31%,23%,and 23%,respectively.Multivariate analysis showed that high PPP2R3A expression(hazard ratio=2.900,95%confidence interval:1.411–5.960,P=0.004)was an independent survival risk factor of HCC patients after LT,and it was also an independent predictor of postoperative tumor recurrence.This study also showed in patients with AFP≥400 ng/mL,the overall survival(P≤0.001)and recurrencefree survival(P=0.023)of those with high PPP2R3A expression were significantly worse compared to those with low PPP2R3A expression.When PPP2R3A expression was low,the overall survival rate(P=0.461)or recurrence-free survival rate(P=0.072)after LT in patients with AFP<400 ng/mL and≥400 ng/mL was not significantly difference.The 1,2,and 3 year survival rate of patients with low PPP2R3A expression and AFP<400 ng/mL were 98%,80%,and 69%,respectively,while patients who met Hangzhou criteria had a posttransplant 1,2,and 3 years overall survival rate of 89%,66%,and 55%,respectively.CONCLUSION High expression of PPP2R3A might be a potential marker for predicting poor prognosis of HCC after LT.Combined with serum AFP levels,PPP2R3A might enhance the accuracy of predicting HCC outcome in patients after LT and supplement the efficacy of the Hangzhou criteria.展开更多
A gene encoding a novel G protein β subunit of β1 subclass, GβMmed was isolated from Microplitis mediator (Hymenoptera: Braconidae). The full-length sequence of GβMmed is 1 119 bp, the cDNA contains a 1 023 bp...A gene encoding a novel G protein β subunit of β1 subclass, GβMmed was isolated from Microplitis mediator (Hymenoptera: Braconidae). The full-length sequence of GβMmed is 1 119 bp, the cDNA contains a 1 023 bp open reading frame that encodes a protein with 340 amino acids, and the predicted molecular weight of GβMmed is 37.23 kDa and isoelectric point is 5.86. By the quantitative real-time RT-PCR method, the tissue-specific expression and quantitative changes in the developmental expression profile of GβMmed were detected. It was found that GβMmed was abundantly expressed in M. mediator antennae, head (without antennae), thorax, abdomen, legs and the wings, and especially at high levels in abdomen. In antennae, expression varied through 1st day before emergence to 5-d-old adults, and had equal expression levels detected in females and males in total. In head, GβMmed expresses while initially high in females, and have another peaked in stage 4 and 1st day, in males showed a peak of GβMmed expression prior to emergence and relatively low levels after emergence. In female abdomen GβMmed expression levels have two peaks in stage 1 and the 5th d, but just have one peak in male abdomen in stage 1. In all other tissues expression was low and stable.展开更多
The western flower thrips(WFT;Frankliniella occidentalis)is a mesophyll cell feeder that damages many crops.Management of WFT is complex due to factors such as high fecundity,short reproduction time,ability to feed on...The western flower thrips(WFT;Frankliniella occidentalis)is a mesophyll cell feeder that damages many crops.Management of WFT is complex due to factors such as high fecundity,short reproduction time,ability to feed on a broad range of host plants,and broad pesticide resistance.These challenges have driven research into developing alternative pest control approaches for WFT.This study analyzed the feasibility of a biological control-based strategy to manage WFT using RNA interference(RNAi)-mediated silencing of WFT endogenous genes.For the delivery of RNAi,we developed transgenic tomato lines expressing double-stranded RNA(dsRNA)of coatomer protein subunit epsilon(CopE)and Toll-like receptor 6(TLR6)from WFT.These genes are involved in critical biological processes of WFT,and their dsRNA can be lethal to these insects when ingested orally.Adult WFT that fed on the transgenic dsRNAexpressing tomato flower stalk showed increased mortality compared with insects that fed on wild-type samples.In addition,WFT that fed on TLR6 and CopE transgenic tomato RNAi lines showed reduced levels of endogenous CopE and TLR6 transcripts,suggesting that their mortality was likely due to RNAi-mediated silencing of these genes.Thus,our findings demonstrate that transgenic tomato plants expressing dsRNA of TLR6 and CopE can be lethal to F.occidentalis,suggesting that these genes may be deployed to control insecticide-resistant WFT.展开更多
Grain size is an important determinant of grain yield in rice.Although dozens of grain size genes have been reported,the molecular mechanisms that control grain size remain to be fully clarified.Here,we report the clo...Grain size is an important determinant of grain yield in rice.Although dozens of grain size genes have been reported,the molecular mechanisms that control grain size remain to be fully clarified.Here,we report the cloning and characterization of GR5(GRAIN ROUND 5),which is allelic to SMOS1/SHB/RLA1/NGR5 and en-codes an AP2 transcription factor.GR5 acts as a transcriptional activator and determines grain size by influencing cell proliferation and expansion.We demonstrated that GR5 physically interacts withfive Gg subunit proteins(RGG1,RGG2,DEP1,GS3,and GGC2)and acts downstream of the G protein complex.Four downstream target genes of GR5 in grain development(DEP2,DEP3,DRW1,and CyCD5;2)were re-vealed and their core T/CGCAC motif identified by yeast one-hybrid,EMSA,and ChIP–PCR experiments.Our results revealed that GR5 interacts with Gg subunits and cooperatively determines grain size by regu-lating the expression of downstream target genes.Thesefindings provide new insight into the genetic reg-ulatory network of the G protein signaling pathway in the control of grain size and provide a potential target for high-yield rice breeding.展开更多
A preliminary study on the interaction of G protein (guanine triphosphate binding pro- tein) b1g2 subunits and their coupled components in cell signal transduction was conducted in vitro. The insect cell lines, Sf9 (S...A preliminary study on the interaction of G protein (guanine triphosphate binding pro- tein) b1g2 subunits and their coupled components in cell signal transduction was conducted in vitro. The insect cell lines, Sf9 (Spodoptera frugiperda) and H5 (Trichoplusia ni ) were used to express the recombinant protein Gb1g2. The cell membrane containing Gb1g2 was isolated through affinity chromatography column with Ni-NTA agarose by FPLC method, and the highly purified protein was obtained. The adenylyl cyclase 2 (AC2) activity assay showed that the purified Gb1g2 could signifi-cantly stimulate AC2 activity. The interaction of b1g2 subunits of G protein with the cytoplasmic tail of various mammalian adenylyl cyclases was monitored by BIAcore technology using NTA sensor chip, which relies on the phenomenon of surface plasmon resonance (SPR). The experiments showed the direct binding of Gb1g2 to the cytoplasmic tail C2 domain of AC2. The specific binding domain of AC2 with Gb1g2 was the same as AC2 activity domain which was stimulated by Gb1g2.展开更多
Bushen Tiansui decoction is composed of six traditional Chinese medicines:Herba Epimedii,Radix Polygoni multiflori,Plastrum testudinis,Fossilia Ossis Mastodi,Radix Polygalae,and Rhizoma Acorus tatarinowii.Because Bus...Bushen Tiansui decoction is composed of six traditional Chinese medicines:Herba Epimedii,Radix Polygoni multiflori,Plastrum testudinis,Fossilia Ossis Mastodi,Radix Polygalae,and Rhizoma Acorus tatarinowii.Because Bushen Tiansui decoction is effective against amyloid beta(Aβ) toxicity,we hypothesized that it would reduce hippocampal synaptic damage and improve cognitive function in Alzheimer's disease.To test this hypothesis,we used a previously established animal model of Alzheimer's disease,that is,microinjection of aggregated Aβ25–35 into the bilateral brain ventricles of Sprague-Dawley rats.We found that long-term(28 days) oral administration of Bushen Tiansui decoction(0.563,1.688,and 3.375 g/m L;4 m L/day) prevented synaptic loss in the hippocampus and increased the expression levels of synaptic proteins,including postsynaptic density protein 95,the N-methyl-D-aspartate receptor 2 B subunit,and Shank1.These results suggested that Bushen Tiansui decoction can protect synapses by maintaining the expression of these synaptic proteins.Bushen Tiansui decoction also ameliorated measures reflecting spatial learning and memory deficits that were observed in the Morris water maze(i.e.,increased the number of platform crossings and the amount of time spent in the target quadrant and decreased escape latency) following intraventricular injections of aggregated Aβ25–35 compared with those measures in untreated Aβ_(25–35)-injected rats.Overall,these results provided evidence that further studies on the prevention and treatment of dementia with this traditional Chinese medicine are warranted.展开更多
Pim kinases contribute to tumor formation and development of lymphoma,which shows enhanced DNA replication,DNA recombination and repair.Endothelial cells (ECs) express all the three members of Pim kinase gene family.W...Pim kinases contribute to tumor formation and development of lymphoma,which shows enhanced DNA replication,DNA recombination and repair.Endothelial cells (ECs) express all the three members of Pim kinase gene family.We hypothesized that DNA repair gene would regulate Pim ex-pression in ECs.Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium.The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining.The siRNA fragments were synthesized and transfected by using Lipofectamine LTX.The total cellular RNA was extracted from the cells by using Trizol reagent.cDNAs were quantified by semi-quantity PCR.The effects of LY294002 and wortmannin on RNA stability in ECs were also ex-amined.Our data showed that LY294002 and wortmannin,phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors,increased Pim mRNA expression in ECs without altering the mRNA stabil-ity.RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1,respectively.Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs.But etoposide,a nucleoside analogue,which could activate DNA-PKcs and ATM,increased Pim expression in ECs.Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.展开更多
Radiotherapy is a part of the front-line treatment regime for many cancers. The mechanisms of radiation-induced effects in cancers mainly involves double-strand breaks (DBS) which plays very important role in maintain...Radiotherapy is a part of the front-line treatment regime for many cancers. The mechanisms of radiation-induced effects in cancers mainly involves double-strand breaks (DBS) which plays very important role in maintaining the stability of gene. As DNA repair gene breast cancer 1 (BRCA1) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) can act to maintain genetic stability though two distinct and complementary mechanisms for DNA DSB repair-homologous recombination (HR) and non-homologous end joining (NHEJ). Therefor, BRCA1 and DNA-PKcs are closely associated with radiation sensitivity, which means that they may be used as a useful tool to predict radio sensitivity in human tumour cells.展开更多
Reversible phosphorylation and dephosphorylation play important roles in cell function and cell signal transduction. PPP2R5A (protein phosphatase 2 regulatory subunit B’ alpha) is responsible for specifically regulat...Reversible phosphorylation and dephosphorylation play important roles in cell function and cell signal transduction. PPP2R5A (protein phosphatase 2 regulatory subunit B’ alpha) is responsible for specifically regulating the catalytic function, substrate specificity and intracellular localization of the tumor suppressor phosphatase PP2A (serine/threonine protein phosphatase 2A). Therefore, the abnormal expression and function of PPP2R5A may be related to canceration. The aim of this study was to reveal its role in the occurrence and development of hepatocellular carcinoma (HCC). It is hoped that the results of this study can provide guidance for the prevention and treatment of HCC. The results showed that PPP2R5A inhibited the proliferation and metastasis of HCC cells, and acted as a tumor suppressor in HCC cells, but it had no significant effect on cell cycle. Further research found that PPP2R5A exerted tumor suppressor efficacy by inhibiting the MAPK/AKT/WNT signaling pathway. Combined with analysis of clinical tissue samples and TCGA database, it was found that the expression of PPP2R5A in tumor tissues of Chinese HCC patients was down-regulated and significantly correlated with the progression-free survival (PFS) of HCC patients. On the contrary, PPP2R5A showed an up-regulation trend in HCC cases in TCGA database although its effect on PFS was the same with that in Chinese HCC patients. Hepatitis B virus (HBV) infection is the main pathogenic factor of HCC in China. It was found that HBV infection reduced the content of PPP2R5A in cells. It was concluded that HBV inhibited the initiation of the protective mechanism mediated by PPP2R5A, making the occurrence and progress of HCC more “unimpeded”. This conclusion will further reveal the role of PPP2R5A in HBV-induced and HBV-unrelated HCC, therefore, providing clues for the prevention and treatment of the two types of HCC, respectively.展开更多
Objective:To examine the frequency of C825T G protein β3 subunit gene polymorphism and its association with obesity of ethnic Kyrgyz.Methods:The study enrolled 210 people,89 patients(35 females,54 males)with obesity(...Objective:To examine the frequency of C825T G protein β3 subunit gene polymorphism and its association with obesity of ethnic Kyrgyz.Methods:The study enrolled 210 people,89 patients(35 females,54 males)with obesity(BMI≥30 kg/m2)and 121 practically healthy patients(38 females,83 males)with normal body weight and no signs of type 2 diabetes(group of control),who were not observed before by a cardiologist.The blood pressure,anthropometry,glucose and lipid profile were examined among all subjects.Genomic DNA was extracted from peripheral blood cells.G proteinβ3 subunit C825T polymorphism was determined by polymerase chain reaction(PCR).Results:TT and CT genotypes carriers were grouped together in one group because the TT genotype was rare.CT+TT genotype frequency in the group with obesity made 0.72 and was significantly higher than that in the control group-0.52(χ2-8.44;P=0.004;odds ratio-2.55;95%CI 1.31-4.23).The statistical analysis revealed that hypertension(45%vs.31.3%,P=0.049)and obesity(51.2%vs.30%,P<0.01)occurred significantly more often in CT+TT genotype carriers than in the CC homozygotes.The results of the multivariate logistic regression analysis showed that the presence of 825T allele(exp β-2.89;95%CI 1.25-6.7;P=0.013),along with the occasional consumption of vegetables(exp β-3.47;95%CI 1.52-7.94;P=0.003)was the significant risk factor for obesity,regardless of gender,age and level of physical activity.In the construction of the similar regression model for hypertension,the statistically significant role of 825T allele was lost after adjustment for obesity as an independent variable.Conclusion:G protein β3 subunit gene C825T allele in the Kyrgyz ethnic group has an association with obesity.展开更多
Colorectal cancer(CRC)is a type of malignant tumor that seriously threatens human health and life,and its treatment has always been a difficulty and hotspot in research.Herein,this study for the first time reports tha...Colorectal cancer(CRC)is a type of malignant tumor that seriously threatens human health and life,and its treatment has always been a difficulty and hotspot in research.Herein,this study for the first time reports that antipsychotic aripiprazole(Ari)against the proliferation of CRC cells both in vitro and in vivo,but with less damage in normal colon cells.Mechanistically,the results showed that5-hydroxytryptamine 2B receptor(HTR2B)and its coupling protein G protein subunit alpha q(Gaq)were highly distributed in CRC cells.Ari had a strong affinity with HTR2B and inhibited HTR2B downstream signaling.Blockade of HTR2B signaling suppressed the growth of CRC cells,but HTR2B was not found to have independent anticancer activity.Interestingly,the binding of Gaq to HTR2B was decreased after Ari treatment.Knockdown of Gaq not only restricted CRC cell growth,but also directly affected the antiCRC efficacy of Ari.Moreover,an interaction between Ari and Gaq was found in that the mutation at amino acid 190 of Gaq reduced the efficacy of Ari.Thus,these results confirm that Gaq coupled to HTR2B was a potential target of Ari in mediating CRC proliferation.Collectively,this study provides a novel effective strategy for CRC therapy and favorable evidence for promoting Ari as an anticancer agent.展开更多
In this study,we investigated the transcriptional spatio-temporal dynamics of the taste 1 receptor(T1R)gene family repertoire in seabream(Sparus aurata[sa]),during larval ontogeny and in adult tissues.In early larval ...In this study,we investigated the transcriptional spatio-temporal dynamics of the taste 1 receptor(T1R)gene family repertoire in seabream(Sparus aurata[sa]),during larval ontogeny and in adult tissues.In early larval development,sa T1R expression arises heterochronously,i.e.the extraoral taste-related perception in the gastrointestinal tract(GIT)anticipates first exogenous feeding(at 9 days post hatching[dph]),followed by the buccal/intraoral perception from 14 dph onwards,supporting the hypothesis that the early onset of the molecular machinery underlying sa T1R expression in the GIT is not induced by food but rather genetically hardwired.During adulthood,we characterized the expression patterns of sa T1R within specific tissues(n=4)distributed in oropharingeal,GIT and brain regions substantiating their functional versatility as chemosensory signaling players to a variety of biological functions beyond oral taste sensation.Further,we provided for the first time direct evidences in fish for m RNA coexpression of a subset of sa T1R genes(mostly sa T1R3,i.e.the common subunit of the heterodimeric T1R complexes for the detection of“sweet”and“umami”substances),with the selected gut peptides ghrelin(ghr),cholecystokinin(cck),hormone peptide yy(pyy)and proglucagon(pg).Each peptide defines the enteroendocrine cells(ECCs)identity,and establishes on morphological basis,a direct link for T1R chemosensing in the regulation of fish digestive processes.Finally,we analyzed the spatial gene expression patterns of 2 taste signaling components functionally homologous to the mammalian G(i)a subunit gustducin,namely sa G(i)a1 and sa G(i)a2,and demonstrated their co-localization with the sa T1R3in EECs,thus validating their direct involvement in taste-like transduction mechanisms of the fish GIT.In conclusion,data provide new insights in the evolutionary conservation of gut sensing in fish suggesting a conserved role for nutrient sensors modulating entero-endocrine secretion.展开更多
基金supported by the Provincial Prize Fund for Distinguished Young and Middle-aged Scientists of Shandong Province(No.BS2011SW053)State Key Laboratory of Plant Cell and Chromosome Engineering(No.PCCE-KF-2014-01)State Key Laboratory of Crop Biology(No.2015KF06)
文摘Wide hybridization is an effective approach for enhancing the resistance of bread wheat (Triticum aestivum L.) to biotic and abiotic stresses by introducing favorable alien genes (Sepsi et al., 2008). Wheatgrass, Thinopyrum intermedium (Host) Barkworth & D.R. Dewey or Agropyron intermedium (Host) Beauvoir (2n = 42; genome formula JJjSjSstst), is a perennial species in the tribe Triticeae and an important source of wheat improvement for biotic and abiotic stress resistance and quality-related traits, such as high grain protein concentration (Chen et al., 1998; 2001; 2003; Han et al., 2004; Li and Wang, 2009). In addition, the ready crossing ability of wheatgrass with various Triticum species has made it popular in germ- plasm development.
文摘BACKGROUND A series of long non-coding RNAs(lncRNAs)have been reported to play a crucial role in cancer biology.Some previous studies report that lncRNA CDKN2B-AS1 is involved in some human malignancies.However,its role in hepatocellular carcinoma(HCC)has not been fully deciphered.AIM To decipher the role of CDKN2B-AS1 in the progression of HCC.METHODS CDKN2B-AS1 expression in HCC was detected by quantitative real-time polymerase chain reaction.The malignant phenotypes of Li-7 and SNU-182 cells were detected by the CCK-8 method,EdU method,and flow cytometry,respectively.RNA immunoprecipitation was executed to confirm the interaction between CDKN2B-AS1 and E2F transcription factor 1(E2F1).Luciferase reporter assay and chromatin immunoprecipitation were performed to verify the binding of E2F1 to the promoter of G protein subunit alpha Z(GNAZ).E2F1 and GNAZ were detected by western blot in HCC cells.RESULTS In HCC tissues,CDKN2B-AS1 was upregulated.Depletion of CDKN2B-AS1 inhibited the proliferation of HCC cells,and the depletion of CDKN2B-AS1 also induced cell cycle arrest and apoptosis.CDKN2B-AS1 could interact with E2F1.Depletion of CDKN2B-AS1 inhibited the binding of E2F1 to the GNAZ promoter region.Overexpression of E2F1 reversed the biological effects of depletion of CDKN2B-AS1 on the malignant behaviors of HCC cells.CONCLUSION CDKN2B-AS1 recruits E2F1 to facilitate GNAZ transcription to promote HCC progression.
基金grants of the National High Technology Research and Development Program,grants of the National Eleventh Five-year Plan Program from the Ministry of Science and Technology of China,Beijing Natural Science Foundation
文摘Objective To explore the association between the three polymorphisms [ C825T, C1429T and G(-350)A] of the gene encoding the G protein beta 3 subunit (GNB3) and hypertension by performing a case-control study in the northern Han Chinese population. Methods We recnaited 731 hypertensive patients and 673 control subjects (the calculated power value was 〉 0.8). Genotyping was performed to identify C825T, C1429T and G(-350)A polymorphisms using the TaqMan assay. Comparisons of allelic and genotypic frequencies between cases and controls were made by using the chi-square test. Logistic regression analyses were performed to investigate the relationships between the three polymorphisms of GNB3 gene under different genetic models (additive, dominant and recessive models). Results The genotype dis- tribution and allele frequencies of C825T, C1429T and G(-350)A polymorphisms did not differ significantly between hypertensive patients and control subjects, either when the full sample was assessed, or when the sample was stratified by gender. No significant association was observed between C825T, C 1429T and G(-350)A polymorphisms and the risk of essential hypertension in any genetic model. Linkage dis- equilibrium was only detected between C825T and C 1429T polymorphisms. Haplotype analyses observed that none of the three estimated haplotypes significantly increased the risk of hypertension. Conclusions Our study suggested that the GNB3 gene polymorphisms [C825T, C 1429T and G(-350)A] were not significantly associated with essential hypertension in northern Han Chinese population.
文摘The cDNA encoding the G protein αq subunit was isolated from the antennae of Helicoverpa assulta (Guen6e) by reverse transcription polymerase chain reaction (RT-PCR) and named as HassGαq. Sequencing analysis showed that the fulllength of HassGαq open reading frame (ORF) is 1 062 bp, 353 amino acid residues are encoded. The predicted molecular weights (MW) and isoelectric point (PI) are 41.5 kD and 5.15, respectively. HassGαq gene was then constructed into expression vector pGEX-4T-2 for over expression in prokaryotic cells. The SDS-PAGE and Western blot analysis showed that induced by Isopropyl-β-D-Thiogalactoside (IPTG), the GST-HassGαq fusion protein is expressed in Escherichia coil BL21, and its MW was found to be about 66 kD nearly equal to the predicted. In addition, RT-PCR analysis showed that the expressions of HassGαq are not tissue specific.
文摘Benzene is an established leukotoxin and leukemogen in humans. We have previously re- ported that exposure of workers to benzene and to benzene metabolite hydroquinone in cultured cells induced DNA-dependent protein kinase catalytic subunit (DNA-PKcs) to mediate the cellular response to DNA double strand break (DSB) caused by DNA-damaging metabolites. In this study, we used a new, small molecule, a selective inhibitor of DNA-PKcs, 2-(morpholin-4-yl)-benzo[h]chomen-4-one (NU7026), as a probe to analyze the molecular events and pathways in hydroquinone-induced DNA DSB repair and apoptosis. Inhibition of DNA-PKcs by NU7026 markedly potentiated the apoptotic and growth inhibitory effects of hydroquinone in proerythroid leukemic K562 cells in a dose-dependent manner. Treatment with NU7026 did not alter the production of reactive oxygen species and oxidative stress by hydroquinone but repressed the protein level of DNA-PKcs and blocked the induction of the kinase mRNA and protein expression by hydroquinone. Moreover, hydroquinone increased the phos- phorylation of Akt to activate Akt, whereas co-treatment with NU7026 prevented the activation of Akt by hydroquinone. Lastly, hydroquinone and NU7026 exhibited synergistic effects on promoting apop- tosis by increasing the protein levels of pro-apoptotic proteins Bax and caspase-3 but decreasing the protein expression of anti-apoptotic protein Bcl-2. Taken together, the findings reveal a central role of DNA-PKcs in hydroquinone-induced hematotoxicity in which it coordinates DNA DSB repair, cell cycle progression, and apoptosis to regulate the response to hydroquinone-induced DNA damage.
基金National Natural Science Foundation of China,No.81372595.
文摘BACKGROUND Protein phosphatase 2 regulatory subunit B''alpha(PPP2R3A)gene has been reported in other tumors,but the influence of PPP2R3A gene expression on the occurrence,development,and prognosis of hepatocellular carcinoma(HCC)remains unclear.AIM To investigate whether the PPP2R3A gene could be used to predict tumor recurrence and survival of HCC patients after liver transplantation(LT).METHODS Diseased liver tissues of HCC patients after LT were collected as well as their clinical data and follow-up information.The immunohistochemical method was used to detect the expression of PPP2R3A protein in the tissues of 108 patients with primary liver cancer.Theχ2 test was used to analyze the relationship between PPP2R3A protein expression levels and the clinicopathological features of tumors.The Kaplan-Meier method was used to analyze overall postoperative survival.The COX proportional hazard model was used to analyze adverse prognostic factors.RESULTS Immunohistochemistry showed that the PPP2R3A protein was mainly expressed in the cytoplasm of HCC cells.Compared to corresponding peritumoral tissues,expression was higher in HCC tissues(P≤0.001).Correlation analysis showed that high PPP2R3A expression was correlated with preoperative serum alphafetoprotein(AFP)levels(P=0.003),tumor-node-metastasis-t stage(P≤0.001),and envelope invasion(P=0.001).Univariate analysis showed that overall survival(P≤0.001)and recurrence-free survival(P=0.025)of patients with high PPP2R3A expression(≥4 points)were poor compared to those with low expression(<4 points).The overall survival rates or recurrence-free survival rates at 1,2,and 3 years with high PPP2R3A expression were 73%,38%,and 23%or 31%,23%,and 23%,respectively.Multivariate analysis showed that high PPP2R3A expression(hazard ratio=2.900,95%confidence interval:1.411–5.960,P=0.004)was an independent survival risk factor of HCC patients after LT,and it was also an independent predictor of postoperative tumor recurrence.This study also showed in patients with AFP≥400 ng/mL,the overall survival(P≤0.001)and recurrencefree survival(P=0.023)of those with high PPP2R3A expression were significantly worse compared to those with low PPP2R3A expression.When PPP2R3A expression was low,the overall survival rate(P=0.461)or recurrence-free survival rate(P=0.072)after LT in patients with AFP<400 ng/mL and≥400 ng/mL was not significantly difference.The 1,2,and 3 year survival rate of patients with low PPP2R3A expression and AFP<400 ng/mL were 98%,80%,and 69%,respectively,while patients who met Hangzhou criteria had a posttransplant 1,2,and 3 years overall survival rate of 89%,66%,and 55%,respectively.CONCLUSION High expression of PPP2R3A might be a potential marker for predicting poor prognosis of HCC after LT.Combined with serum AFP levels,PPP2R3A might enhance the accuracy of predicting HCC outcome in patients after LT and supplement the efficacy of the Hangzhou criteria.
基金support from the Na-tional Natural Science Foundation of China (30871640,30330410)the National Basic Research Program ofChina (2007CB109202)the Research Foundationof State Key Laboratory for Biology of Plant Diseasesand Insect Pests of China (SKL2007SR01)
文摘A gene encoding a novel G protein β subunit of β1 subclass, GβMmed was isolated from Microplitis mediator (Hymenoptera: Braconidae). The full-length sequence of GβMmed is 1 119 bp, the cDNA contains a 1 023 bp open reading frame that encodes a protein with 340 amino acids, and the predicted molecular weight of GβMmed is 37.23 kDa and isoelectric point is 5.86. By the quantitative real-time RT-PCR method, the tissue-specific expression and quantitative changes in the developmental expression profile of GβMmed were detected. It was found that GβMmed was abundantly expressed in M. mediator antennae, head (without antennae), thorax, abdomen, legs and the wings, and especially at high levels in abdomen. In antennae, expression varied through 1st day before emergence to 5-d-old adults, and had equal expression levels detected in females and males in total. In head, GβMmed expresses while initially high in females, and have another peaked in stage 4 and 1st day, in males showed a peak of GβMmed expression prior to emergence and relatively low levels after emergence. In female abdomen GβMmed expression levels have two peaks in stage 1 and the 5th d, but just have one peak in male abdomen in stage 1. In all other tissues expression was low and stable.
基金supported by the Basic Science Research Program through the National Research Foundation(NRF),Ministry of Education,Korea(2021R1I1A1A01041938)a grant from the New Breeding Technologies Development Program,Rural Development Administration,Korea(PJ0165432022)supported in part by the BK21 Plus Program,Ministry of Education,Korea。
文摘The western flower thrips(WFT;Frankliniella occidentalis)is a mesophyll cell feeder that damages many crops.Management of WFT is complex due to factors such as high fecundity,short reproduction time,ability to feed on a broad range of host plants,and broad pesticide resistance.These challenges have driven research into developing alternative pest control approaches for WFT.This study analyzed the feasibility of a biological control-based strategy to manage WFT using RNA interference(RNAi)-mediated silencing of WFT endogenous genes.For the delivery of RNAi,we developed transgenic tomato lines expressing double-stranded RNA(dsRNA)of coatomer protein subunit epsilon(CopE)and Toll-like receptor 6(TLR6)from WFT.These genes are involved in critical biological processes of WFT,and their dsRNA can be lethal to these insects when ingested orally.Adult WFT that fed on the transgenic dsRNAexpressing tomato flower stalk showed increased mortality compared with insects that fed on wild-type samples.In addition,WFT that fed on TLR6 and CopE transgenic tomato RNAi lines showed reduced levels of endogenous CopE and TLR6 transcripts,suggesting that their mortality was likely due to RNAi-mediated silencing of these genes.Thus,our findings demonstrate that transgenic tomato plants expressing dsRNA of TLR6 and CopE can be lethal to F.occidentalis,suggesting that these genes may be deployed to control insecticide-resistant WFT.
基金supported by the National Key Research and Development Program of China (2022YFF1002901)the Hainan Yazhou Bay Seed Laboratory (B21HJ0215)+4 种基金the Natural Science Foundation of China (32272109)the Nanfan special project (ZDXM2315)of CAASthe Natural Science Foundation of China (32072048,U2004204)the Qian Qian Academician Workstation,the specific research fund of the Innovation Platform for Academicians of Hainan Province (YSPTZX202303)the Key Research and Development Program of Zhejiang Province (2021C02056).
文摘Grain size is an important determinant of grain yield in rice.Although dozens of grain size genes have been reported,the molecular mechanisms that control grain size remain to be fully clarified.Here,we report the cloning and characterization of GR5(GRAIN ROUND 5),which is allelic to SMOS1/SHB/RLA1/NGR5 and en-codes an AP2 transcription factor.GR5 acts as a transcriptional activator and determines grain size by influencing cell proliferation and expansion.We demonstrated that GR5 physically interacts withfive Gg subunit proteins(RGG1,RGG2,DEP1,GS3,and GGC2)and acts downstream of the G protein complex.Four downstream target genes of GR5 in grain development(DEP2,DEP3,DRW1,and CyCD5;2)were re-vealed and their core T/CGCAC motif identified by yeast one-hybrid,EMSA,and ChIP–PCR experiments.Our results revealed that GR5 interacts with Gg subunits and cooperatively determines grain size by regu-lating the expression of downstream target genes.Thesefindings provide new insight into the genetic reg-ulatory network of the G protein signaling pathway in the control of grain size and provide a potential target for high-yield rice breeding.
基金This work was sup-ported by the National Natural Science Foundation of China (Grant No. 30170615) and National Major Basis Study Develop-mental Plan 973 Project (TG2000016208).
文摘A preliminary study on the interaction of G protein (guanine triphosphate binding pro- tein) b1g2 subunits and their coupled components in cell signal transduction was conducted in vitro. The insect cell lines, Sf9 (Spodoptera frugiperda) and H5 (Trichoplusia ni ) were used to express the recombinant protein Gb1g2. The cell membrane containing Gb1g2 was isolated through affinity chromatography column with Ni-NTA agarose by FPLC method, and the highly purified protein was obtained. The adenylyl cyclase 2 (AC2) activity assay showed that the purified Gb1g2 could signifi-cantly stimulate AC2 activity. The interaction of b1g2 subunits of G protein with the cytoplasmic tail of various mammalian adenylyl cyclases was monitored by BIAcore technology using NTA sensor chip, which relies on the phenomenon of surface plasmon resonance (SPR). The experiments showed the direct binding of Gb1g2 to the cytoplasmic tail C2 domain of AC2. The specific binding domain of AC2 with Gb1g2 was the same as AC2 activity domain which was stimulated by Gb1g2.
基金supported by the National Natural Science Foundation of China,No.81373705the Natural Science Foundation of Hunan Province in China,No.13JJ3030
文摘Bushen Tiansui decoction is composed of six traditional Chinese medicines:Herba Epimedii,Radix Polygoni multiflori,Plastrum testudinis,Fossilia Ossis Mastodi,Radix Polygalae,and Rhizoma Acorus tatarinowii.Because Bushen Tiansui decoction is effective against amyloid beta(Aβ) toxicity,we hypothesized that it would reduce hippocampal synaptic damage and improve cognitive function in Alzheimer's disease.To test this hypothesis,we used a previously established animal model of Alzheimer's disease,that is,microinjection of aggregated Aβ25–35 into the bilateral brain ventricles of Sprague-Dawley rats.We found that long-term(28 days) oral administration of Bushen Tiansui decoction(0.563,1.688,and 3.375 g/m L;4 m L/day) prevented synaptic loss in the hippocampus and increased the expression levels of synaptic proteins,including postsynaptic density protein 95,the N-methyl-D-aspartate receptor 2 B subunit,and Shank1.These results suggested that Bushen Tiansui decoction can protect synapses by maintaining the expression of these synaptic proteins.Bushen Tiansui decoction also ameliorated measures reflecting spatial learning and memory deficits that were observed in the Morris water maze(i.e.,increased the number of platform crossings and the amount of time spent in the target quadrant and decreased escape latency) following intraventricular injections of aggregated Aβ25–35 compared with those measures in untreated Aβ_(25–35)-injected rats.Overall,these results provided evidence that further studies on the prevention and treatment of dementia with this traditional Chinese medicine are warranted.
基金supported by grants from the National Natural Science Foundation of China (No. 30900631)Hubei Provincial Department of Education (No. Q20102101 andD20082406)
文摘Pim kinases contribute to tumor formation and development of lymphoma,which shows enhanced DNA replication,DNA recombination and repair.Endothelial cells (ECs) express all the three members of Pim kinase gene family.We hypothesized that DNA repair gene would regulate Pim ex-pression in ECs.Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium.The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining.The siRNA fragments were synthesized and transfected by using Lipofectamine LTX.The total cellular RNA was extracted from the cells by using Trizol reagent.cDNAs were quantified by semi-quantity PCR.The effects of LY294002 and wortmannin on RNA stability in ECs were also ex-amined.Our data showed that LY294002 and wortmannin,phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors,increased Pim mRNA expression in ECs without altering the mRNA stabil-ity.RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1,respectively.Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs.But etoposide,a nucleoside analogue,which could activate DNA-PKcs and ATM,increased Pim expression in ECs.Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.
文摘Radiotherapy is a part of the front-line treatment regime for many cancers. The mechanisms of radiation-induced effects in cancers mainly involves double-strand breaks (DBS) which plays very important role in maintaining the stability of gene. As DNA repair gene breast cancer 1 (BRCA1) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) can act to maintain genetic stability though two distinct and complementary mechanisms for DNA DSB repair-homologous recombination (HR) and non-homologous end joining (NHEJ). Therefor, BRCA1 and DNA-PKcs are closely associated with radiation sensitivity, which means that they may be used as a useful tool to predict radio sensitivity in human tumour cells.
文摘Reversible phosphorylation and dephosphorylation play important roles in cell function and cell signal transduction. PPP2R5A (protein phosphatase 2 regulatory subunit B’ alpha) is responsible for specifically regulating the catalytic function, substrate specificity and intracellular localization of the tumor suppressor phosphatase PP2A (serine/threonine protein phosphatase 2A). Therefore, the abnormal expression and function of PPP2R5A may be related to canceration. The aim of this study was to reveal its role in the occurrence and development of hepatocellular carcinoma (HCC). It is hoped that the results of this study can provide guidance for the prevention and treatment of HCC. The results showed that PPP2R5A inhibited the proliferation and metastasis of HCC cells, and acted as a tumor suppressor in HCC cells, but it had no significant effect on cell cycle. Further research found that PPP2R5A exerted tumor suppressor efficacy by inhibiting the MAPK/AKT/WNT signaling pathway. Combined with analysis of clinical tissue samples and TCGA database, it was found that the expression of PPP2R5A in tumor tissues of Chinese HCC patients was down-regulated and significantly correlated with the progression-free survival (PFS) of HCC patients. On the contrary, PPP2R5A showed an up-regulation trend in HCC cases in TCGA database although its effect on PFS was the same with that in Chinese HCC patients. Hepatitis B virus (HBV) infection is the main pathogenic factor of HCC in China. It was found that HBV infection reduced the content of PPP2R5A in cells. It was concluded that HBV inhibited the initiation of the protective mechanism mediated by PPP2R5A, making the occurrence and progress of HCC more “unimpeded”. This conclusion will further reveal the role of PPP2R5A in HBV-induced and HBV-unrelated HCC, therefore, providing clues for the prevention and treatment of the two types of HCC, respectively.
文摘Objective:To examine the frequency of C825T G protein β3 subunit gene polymorphism and its association with obesity of ethnic Kyrgyz.Methods:The study enrolled 210 people,89 patients(35 females,54 males)with obesity(BMI≥30 kg/m2)and 121 practically healthy patients(38 females,83 males)with normal body weight and no signs of type 2 diabetes(group of control),who were not observed before by a cardiologist.The blood pressure,anthropometry,glucose and lipid profile were examined among all subjects.Genomic DNA was extracted from peripheral blood cells.G proteinβ3 subunit C825T polymorphism was determined by polymerase chain reaction(PCR).Results:TT and CT genotypes carriers were grouped together in one group because the TT genotype was rare.CT+TT genotype frequency in the group with obesity made 0.72 and was significantly higher than that in the control group-0.52(χ2-8.44;P=0.004;odds ratio-2.55;95%CI 1.31-4.23).The statistical analysis revealed that hypertension(45%vs.31.3%,P=0.049)and obesity(51.2%vs.30%,P<0.01)occurred significantly more often in CT+TT genotype carriers than in the CC homozygotes.The results of the multivariate logistic regression analysis showed that the presence of 825T allele(exp β-2.89;95%CI 1.25-6.7;P=0.013),along with the occasional consumption of vegetables(exp β-3.47;95%CI 1.52-7.94;P=0.003)was the significant risk factor for obesity,regardless of gender,age and level of physical activity.In the construction of the similar regression model for hypertension,the statistically significant role of 825T allele was lost after adjustment for obesity as an independent variable.Conclusion:G protein β3 subunit gene C825T allele in the Kyrgyz ethnic group has an association with obesity.
基金supported by Chongqing basic research and frontier exploration project(cstc2022ycjh-bgzxm0119,China)。
文摘Colorectal cancer(CRC)is a type of malignant tumor that seriously threatens human health and life,and its treatment has always been a difficulty and hotspot in research.Herein,this study for the first time reports that antipsychotic aripiprazole(Ari)against the proliferation of CRC cells both in vitro and in vivo,but with less damage in normal colon cells.Mechanistically,the results showed that5-hydroxytryptamine 2B receptor(HTR2B)and its coupling protein G protein subunit alpha q(Gaq)were highly distributed in CRC cells.Ari had a strong affinity with HTR2B and inhibited HTR2B downstream signaling.Blockade of HTR2B signaling suppressed the growth of CRC cells,but HTR2B was not found to have independent anticancer activity.Interestingly,the binding of Gaq to HTR2B was decreased after Ari treatment.Knockdown of Gaq not only restricted CRC cell growth,but also directly affected the antiCRC efficacy of Ari.Moreover,an interaction between Ari and Gaq was found in that the mutation at amino acid 190 of Gaq reduced the efficacy of Ari.Thus,these results confirm that Gaq coupled to HTR2B was a potential target of Ari in mediating CRC proliferation.Collectively,this study provides a novel effective strategy for CRC therapy and favorable evidence for promoting Ari as an anticancer agent.
基金covered by the National Research Agency(AEI,Spain)(grant number:PID2019-103969RB-C33)to Jos e M.Cerd a-Reverter。
文摘In this study,we investigated the transcriptional spatio-temporal dynamics of the taste 1 receptor(T1R)gene family repertoire in seabream(Sparus aurata[sa]),during larval ontogeny and in adult tissues.In early larval development,sa T1R expression arises heterochronously,i.e.the extraoral taste-related perception in the gastrointestinal tract(GIT)anticipates first exogenous feeding(at 9 days post hatching[dph]),followed by the buccal/intraoral perception from 14 dph onwards,supporting the hypothesis that the early onset of the molecular machinery underlying sa T1R expression in the GIT is not induced by food but rather genetically hardwired.During adulthood,we characterized the expression patterns of sa T1R within specific tissues(n=4)distributed in oropharingeal,GIT and brain regions substantiating their functional versatility as chemosensory signaling players to a variety of biological functions beyond oral taste sensation.Further,we provided for the first time direct evidences in fish for m RNA coexpression of a subset of sa T1R genes(mostly sa T1R3,i.e.the common subunit of the heterodimeric T1R complexes for the detection of“sweet”and“umami”substances),with the selected gut peptides ghrelin(ghr),cholecystokinin(cck),hormone peptide yy(pyy)and proglucagon(pg).Each peptide defines the enteroendocrine cells(ECCs)identity,and establishes on morphological basis,a direct link for T1R chemosensing in the regulation of fish digestive processes.Finally,we analyzed the spatial gene expression patterns of 2 taste signaling components functionally homologous to the mammalian G(i)a subunit gustducin,namely sa G(i)a1 and sa G(i)a2,and demonstrated their co-localization with the sa T1R3in EECs,thus validating their direct involvement in taste-like transduction mechanisms of the fish GIT.In conclusion,data provide new insights in the evolutionary conservation of gut sensing in fish suggesting a conserved role for nutrient sensors modulating entero-endocrine secretion.