Long noncoding RNAs HAND2-AS1 ultrasound microbubbles suppress hepatocellular carcinoma progression by regulating the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis

BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.

Point mutation of 5' noncoding region of BCL-6gene in primary gastric lymphomas

AIM: To investigate the mutations of the 5' noncoding region of BCL-6 gene in Chinese patients with primary gastric lymphomas. METHODS: PCR and direct DNA sequencing were used to identify BCL-6 gene mutations in the 5' noncoding region in 29 cases of gastric diffuse large B-cell lymphoma (DLBCL) and 18 cases of gastric mucosa-associated lymphoid tissue (MALT) lymphoma as well as 10 cases of reactive hyperplasia of lymph node (LRH). RESULTS: Six of 29 gastric DLBCLs (20.7%), 4 of 18 gastric MALT lymphomas (22.2%) and 1 of 10 LRHs(10%) were found to have mutations. All mutations were single-base substitutions and the frequency of single-base changes was 0.20×1O^(-2)-1.02×1O^(-2)per bp. CONCLUSION: Point mutations in the 5' noncoding region of BCL-6 gene are found in Chinese patients with primary gastric DLBCLs and MALT lymphomas, suggesting that they may, in some extent, participate in the pathogenesis of primary gastric DLBCLs and MALT lymphomas.

Long noncoding RNA RP4 functions as a competing endogenous RNA through miR-7-5p sponge activity in colorectal cancer

AIM To investigate the role of long noncoding RNA(lnc RNA) RP4 in colorectal cancer.METHODS Lentivirus-mediated lnc RNA RP4 overexpression and knockdown were performed in the colorectal cancer cell line SW480. Cell proliferation, tumor growth, and early apoptosis were evaluated by a cell counting kit-8 assay, an in vivo xenograft tumor model, and annexin V/propidium iodide staining, respectively. Analysis of the lnc RNA RP4 mechanism involved assessment of the association of its expression with mi R-7-5 p and the SH3 GLB1 gene. Western blot analysis was also performed to assess the effect of lnc RNA RP4 on the autophagy-mediated cell death pathway and phosphatidylinositol-3-kinase(PI3 K)/Akt signaling.RESULTS Cell proliferation, tumor growth, and early apoptosis in SW480 cells were negatively regulated by lnc RNA RP4. Functional experiments indicated that lnc RNA RP4 directly upregulated SH3 GLB1 expression by acting as a competing endogenous RNA(ce RNA) for mi R-7-5 p. This interaction led to activation of the autophagy-mediated cell death pathway and de-repression of PI3 K and Akt phosphorylation in colorectal cancer cells in vivo.CONCLUSION Our results demonstrated that lnc RNA RP4 is a ce RNA that plays an important role in the pathogenesis of colorectal cancer, and could be a potential therapeutic target for colorectal cancer treatment.

长链的非编码RNA生长停滞特异性转录本5在骨关节炎中的研究进展

骨关节炎(OA)是一种以软骨退变、骨重建为主要病理特点的慢性关节疾病。目前没有可治愈该疾病的药物及手段,治疗策略有限的根本原因是对其潜在发病机制的认识不足。长链的非编码RNA(LncRNAs)被认为是许多疾病中的一类新的调控因子,是目前研究OA发病机制的一个重要切入点。LncRNA生长停滞特异性转录本5(Cas5)参与调控细胞的增殖、分化及凋亡,是一种在软骨细胞中特异性高表达的LncRNA。然而,由于LncRNA Cas5作用机制复杂,其与OA相关具体分子机制目前仍不明确。该文针对LncRNA Cas5在OA中的软骨细胞凋亡、软骨基质代谢成骨分化及治疗方面的作用进行综述。

Potential role of long noncoding RNA RP5-881L22.5 as a novel biomarker and therapeutic target of colorectal cancer

BACKGROUND The incidence of colorectal cancer in humans is high,and it is in the top five for cancer-related morbidity and mortality.It is one of the main threats to human health.The function of long noncoding RNAs in tumor occurrence and development has gradually gained attention in recent years.In increasing numbers of studies,researchers have demonstrated that it plays an important role in the pathogenesis of colorectal cancer.AIM To find out if long noncoding RNA RP5-881L22.5 played a role in the pathogenesis of colorectal cancer in relation to the tumor microenvironment.METHODS We analyzed the transcriptome data and clinical data in The Cancer Genome Atlas-colon adenocarcinoma.The CIRBERSORT algorithm was applied to evaluate these tumor-infiltrating immune cells in The Cancer Genome Atlas-colon adenocarcinoma cancer tissue samples.Using the“estimate”package in R,we assessed the tumor immune microenvironment.The expression level of RP5-881L22.5 in tumor tissue and adjacent normal tissue samples from 4 pairs of colorectal cancer patients was determined by quantitative reverse transcription PCR.Colorectal cancer cells were tested for invasiveness using a transwell invasion assay after RP5-881L22.5 expression was knocked down.RESULTS The expression of lncRNA RP5-881L22.5 was related to the clinical characteristics of the tumors,and it was negatively related to the infiltration level of immune cells in the tumor microenvironment and the expression of T cell inhibitory receptors.A major function of its coexpressed mRNA was to regulate tumor immunity,such as the immune response.When quantitative reverse transcription PCR was performed on tumor tissues from 4 pairs of colorectal cancer patients,the results showed that RP5-881L22.5 was highly expressed.Subsequently,knocking down the expression of RP5-881L22.5,the invasiveness of colorectal cancer cell lines was reduced,and the apoptosis rate was increased.CONCLUSION RP5-881L22.5 plays a crucial role in the microenvironment of tumors as well as in the pathogenesis of colorectal cancer.The relationship between RP5-881L22.5 and the tumor immune microenvironment deserves further study.

Study on the Knockout and the Soluble Prokaryotic Expression of VP5 Protein Transmembrane Region of IBDV

[Objective] The research aimed to construct the prokaryotic expression vector of VP5 protein of IBDV.The transmembrane region sequence of VP5 protein was knocked out.Moreover,the expression,separation and purification of objective protein were carried out.[Method] PCR technology was used to respectively amplify the extracellular and intracellular fragments of VP5 gene of IBDV.Then,the two fragments were simultaneously linked to pET-28b（＋）,and it was the vector-intracellular fragment-extracellular fragment-vector.The recombinant expression plasmid pET-VP5-FC and the improved pET-VP5-SC of VP5 whose transmembrane region gene fragment was knocked out were constructed.Then,the expression plasmid was transformed into BL21（DE3）.After IPTG induction,the recombinant protein was purified by Ni affinity chromatography and the gel filtration chromatography.[Result] The soluble expressed VP5 of IBDV was obtained.[Conclusion] The research laid the foundation for further studying the structure and function of VP5 protein.

Three 5’-flanking Regions of crtO Encoding β-carotene Oxygenase in Haematococcus pluvialis

Three separate 5＇-flanking regions （1.1 kb, 1.9 kb and 2.2 kb） of crtO were cloned through walking upstream. Results of sequence analysis show that three 5＇-flanking regions of crtO might have some similar putative cis-acting elements such as ABA （abscisic acid）-responsive element （ABRE）, C-repeat/dehydration responsive element （C-repeat/DRE）, light-responsive element （G-box, GAG-motif, I-box and ATC-motif）, wound-responsive element （WUN-motif）, auxin-responsive element （TGA-element）, MeJA-responsive element （TGACG-element） and MYB binding site （MBS）, except for typical TATA box or CCAAT box. These findings might mean diversiform regulatory patterns of crtO being in astaxanthin biosynthesis of Haematococcus pluvialis.

抑郁模型大鼠不同脑区5-HT_(1A)蛋白表达及加味温胆汤的干预作用

目的:基于不同脑区5-HT_(1A)蛋白表达,探讨模型大鼠抑郁发病机制以及加味温胆汤的干预作用。方法:将32只SPF级雄性SD大鼠(体质量180~200 g)随机分为正常对照组、抑郁模型组、抑郁中药组、抑郁西药组各8只。采用慢性轻度不可预见性应激制作大鼠抑郁模型,其中正常对照组、抑郁模型组大鼠灌胃生理盐水,抑郁中药组灌胃12 g/kg加味温胆汤,抑郁西药组灌胃1.8 mg/kg百忧解。35天后进行行为学检测及取材,通过Western blot法检测各组大鼠杏仁核、前额皮质、海马CA3区及下丘脑组织中5-HT_(1A)蛋白表达。结果:行为学数据显示,抑郁模型组大鼠水平运动次数、垂直运动次数以及总路程较正常对照组减少(P<0.05),抗抑郁西药百忧解只对总路程有影响(P<0.05),而加味温胆汤能改善模型大鼠抑郁行为表现(P<0.05),提高水平运动、垂直运动次数,延长总路程。抑郁模型组大鼠海马CA3区5-HT_(1A)蛋白表达水平升高,加味温胆汤能降低海马CA3区5-HT_(1A)蛋白表达(P<0.05),百忧解能提高杏仁核5-HT_(1A)蛋白表达(P<0.05)。结论:抑郁模型大鼠不同脑区5-HT_(1A)蛋白表达不一致,抑郁模型大鼠海马CA3区5-HT_(1A)蛋白表达升高,加味温胆汤可能通过降低抑郁模型大鼠海马CA3区5-HT_(1A)蛋白表达发挥抗抑郁作用,而百忧解能提高杏仁核5-HT_(1A)蛋白表达,其抗抑郁机制有待进一步阐明。

自拟安神方对心肾不交证失眠大鼠皮质区和海马区5-HT和DA信号通路影响

目的探索自拟安神方(简称安神方)对大鼠不同脑区5-羟色胺(5-hydroxytryptamine,5-HT)和多巴胺(dopamine,DA)信号通路的影响,揭示安神方抗大鼠心肾不交证失眠的机制。方法将SD雄性大鼠随机分为正常组,模型组,阳性对照组,安神方高、中、低剂量组,每组10只。除正常组外,其他各组采用多因素方法制备改良的心肾不交证失眠大鼠模型,灌胃治疗2周。HE染色观察大鼠脑组织形态结构。高效液相色谱-电化学法(HPLC-ECD)检测大鼠皮质区和海马区DA和5-HT的含量。免疫蛋白印迹法检测5-HT信号通路上5-HT_(1A)受体、5-HT_(2A)受体、磷酸腺苷反应元件结合蛋白(cAMP-response element binding protein,CREB)、蛋白激酶A(protein kinase A,PKA)和DA信号通路上D_(1)R_(1)在皮质区和海马区的蛋白表达。结果HE染色显示模型组皮质区神经元有少量的凋亡,海马结构区变化不明显;安神方组皮质区神经元凋亡有所减轻。HPLC-ECD结果显示,与正常组比较,模型组皮质区和海马区DA含量均升高(P<0.05),5-HT含量均显著下降(P<0.05);与模型组比较,安神方组皮质区和海马区DA含量显著降低(P<0.05),同时海马区5-HT含量有所提高(P<0.05)。免疫印迹结果显示,与正常组比较,模型组皮质区5-HT_(1A)和CREB蛋白表达降低(P<0.05),海马区D_(1)R_(1)蛋白表达升高(P<0.05);与模型组比较,安神方组皮质区5-HT_(1A)和信号通路上关键蛋白CREB的蛋白表达上调(P<0.05),并且海马区D_(1)R_(1)的蛋白表达下调(P<0.05),其信号通路上CREB蛋白表达差异虽无统计学意义,但各给药组仍有上调趋势。结论安神方抗心肾不交证失眠的机制可能与抑制皮质区神经元凋亡,并促进皮质区和海马区5-HT通路功能、抑制DA通路功能有关。

Quasispecies evolution in NS5A region of hepatitis C virus genotype 1b during interferon or combined interferon-ribavirin therapy

AIM: To evaluate the implication of substitutions in the hepatitis C virus (HCV) non-structural 5A (NS5A) protein in the resistance of HCV during mono-interferon (IFN) or combined IFN-ribavirin (IFN-R) therapy. Although NS5A has been reported to interact with the HCV RNA- dependent RNA polymerase, NS5B, as well as with many cellular proteins, the function of NS5A in the life cycle of HCV remains unclear. METHODS: HCV quasispecies were studied by clon- ing and sequencing of sequential isolates from patients infected by HCV genotype 1b. Patients were treated by IFN-α2b for 3 mo followed by IFN-α2b alone or com- bined IFN-R therapy for 9 additional months. Patients were categorized intro two groups based on their re- sponse to the treatments: 7 with sustained virological re- sponse (SVR) (quasispecies = 150) and 3 non-respond- ers (NR) to IFN-R (quasispecies = 106). RESULTS: Prior to treatment, SVR patients displayed a lower complexity of quasispecies than NR patients. Most patients had a decrease in the complexity of quasispe- cies during therapy. Analysis of amino acids substitu- tions showed that the degree of the complexity of the interferon sensitivity-determining region (ISDR) and the V3 domain of NS5A protein was able to discriminate thetwo groups of patients. Moreover, SVR patients displayed more variability in the NS5A region than NR patients. CONCLUSION: These results suggest that detailed mo- lecular analysis of the NS5A region may be important for understanding its function in IFN response during HCV 1b infection.

Future Precipitation Extremes in China under Climate Change and Their Physical Quantification Based on a Regional Climate Model and CMIP5 Model Simulations

The atmospheric water holding capacity will increase with temperature according to Clausius-Clapeyron scaling and affects precipitation.The rates of change in future precipitation extremes are quantified with changes in surface air temperature.Precipitation extremes in China are determined for the 21st century in six simulations using a regional climate model,RegCM4,and 17 global climate models that participated in CMIP5.First,we assess the performance of the CMIP5 models and RCM runs in their simulation of extreme precipitation for the current period(RF:1982-2001).The CMIP5 models and RCM results can capture the spatial variations of precipitation extremes,as well as those based on observations:OBS and XPP.Precipitation extremes over four subregions in China are predicted to increase in the mid-future(MF:2039-58)and far-future(FF:2079-98)relative to those for the RF period based on both the CMIP5 ensemble mean and RCM ensemble mean.The secular trends in the extremes of the CMIP5 models are predicted to increase from 2008 to 2058,and the RCM results show higher interannual variability relative to that of the CMIP5 models.Then,we quantify the increasing rates of change in precipitation extremes in the MF and FF periods in the subregions of China with the changes in surface air temperature.Finally,based on the water vapor equation,changes in precipitation extremes in China for the MF and FF periods are found to correlate positively with changes in the atmospheric vertical wind multiplied by changes in surface specific humidity(significant at the p<0.1 level).

Future temperature changes over the critical Belt and Road region based on CMIP5 models

Based on data of 22 models from the Coupled Model Inter-comparison Project Phase 5(CMIP5),the performance of climate simulation is assessed and future changes under RCP2.6,RCP4.5 and RCP8.5 are projected over critical Belt and Road region.Compared with observations,the CMIP5 models simulate the linear trend and spatial distribution of the annual mean surface air temperature(SAT)better in the north(NBR)and south(SBR)of the Belt and Road region.The trend of the 22-model ensemble mean(CMIP5 MME)is 0.70/0.50
C per 100 years from 1901 to 2005,and the observed trend is 1.11/0.77
C per 100 years in the NBR/SBR region.After 1971,the relative error between CMIP5 MME and observations is 22%/15%in the NBR/SBR region.Seven/nine models are selected in the NBR/SBR to project future SAT changes under three RCP scenarios.For 2081e2100,warming in the NBR/SBR is projected to be(1.16±0.29)/(0.72±0.32)
C,(2.41±0.54)/(1.55±0.44)
C,and(5.23±1.02)/(3.33±0.65)
C for RCP2.6,RCP4.5,and RCP8.5,respectively.Under the RCP scenarios,the NBR region shows greater warming than the SBR region.The most significant warming is expected in Kazakhstan and the northern part of the SBR.The associated uncertainty generally increases with time under the three RCP scenarios.Furthermore,increases in warming over the Belt and Road region are more remarkable under higher-emission scenarios than lower-emission ones.

Projected changes in mean and extreme climates over Hindu Kush Himalayan region by 21 CMIP5 models

Based on the outputs from 21 CMIP5 (Coupled Model Intercomparison Project phase 5) models, future changes in the mean temperature, precipitation and four climate extreme indices (annual maximum of daily maximum temperature (TXx), minimum of daily minimum temperature (TNn), annual total precipitation when the daily amount exceeds the 95th percentile of wet-day precipitation (R95p), and maximum consecutive 5-day precipitation (RX5day)) over Hindu Kush Himalayan (HKH) region are investigated under the greenhouse gas concentration pathways of RCP4.5 and RCP8.5. Two periods of the 21st century, 2036e2065 and 2066e2095, are selected, with the reference period is considered as 1976e2005. Results show general increase of the mean temperature, TXx and TNn under both scenarios, with the largest increases found during 2066e2095 under RCP8.5. Future precipitation is projected to increase over most part of HKH, except for the northwestern part. Intensification of the precipitation extremes is projected over the region. The uncertainties of mean temperature, TXx and TNn over the HKH1 subregions are the largest compared to the other three subregions and the overall HKH. Besides RX5day during 2036e2065 over HKH1, the uncertainties of R95p and RX5day tend to be larger following the increase of greenhouse gas concentrations. The multimodel ensemble medians of temperature and four extreme indices under RCP8.5 are projected to be larger than those under RCP4.5 in each of the subregions.

Functional characterization of a Δ6 fatty acid desaturase gene and its 5′-upstream region cloned from the arachidonic acid-rich microalga Myrmecia incisa Reisigl(Chlorophyta)

It is suggested that Δ6 fatty acid desaturase(FAD) plays a critical role in the biosynthesis of polyunsaturated fatty acids in plants and microalgae. But why does it adapt to the changed environments such as nitrogen starvation is seldom understood. One Δ6 FAD gene( MiD6 fad) from an arachidonic acidrich microalga M yrmecia incisa Reisigl(Chlorophyta) was first heterologously expressed in S accharomyces cerevisiae for the identification of function. The fatty acid profile of transgenic yeast detected by gas chromatography-mass spectrometry illustrated that the enzyme MiD6 FAD could convert linoleic and ?-linolenic acids to γ-linolenic and stearidonic acids, respectively, demonstrating that M iD6 fad encoded a Δ6 FAD. A 1 965-bp fragment of the cloned 2 347-bp 5′-upstream region of M iD6 fad was next subcloned and fused upstream with green fluorescent protein(GFP) gene to replace the GAL1 promoter of the vector pYES2. The generated construct was transformed into S. cerevisiae for function determination. Confocal microscopic images of the transformed line illustrated that this inserted fragment could drive GFP expression, which was further verified by fluorescence intensity quantification and Western blot analysis using antiGFP antibody. The conversion efficiency(approximately 2%-3%) of MiD6 FAD was much lower than the reported ? 3 FAD and Δ6 elongase in this microalga, suggesting that MiD6 FAD catalysed the possible ratelimiting step for ArA biosynthesis. The presence of several putative c is-acting regulatory elements in this identified promoter sheds new light on the regulation mechanism research of Δ6 FAD transcription for the ArA production in M. incisa in changing environmental factors.

Long noncoding RNA X-inactive specific transcript regulates NLR family pyrin domain containing 3/caspase-1-mediated pyroptosis in diabetic nephropathy

BACKGROUND NLRP3-mediated pyroptosis is recognized as an essential modulator of renal disease pathology.Long noncoding RNAs(lncRNAs)are active participators of diabetic nephropathy(DN).X inactive specific transcript(XIST)expression has been reported to be elevated in the serum of DN patients.AIM To evaluate the mechanism of lncRNA XIST in renal tubular epithelial cell(RTEC)pyroptosis in DN.METHODS A DN rat model was established through streptozotocin injection,and XIST was knocked down by tail vein injection of the lentivirus LV sh-XIST.Renal metabolic and biochemical indices were detected,and pathological changes in the renal tissue were assessed.The expression of indicators related to inflammation and pyroptosis was also detected.High glucose(HG)was used to treat HK2 cells,and cell viability and lactate dehydrogenase(LDH)activity were detected after silencing XIST.The subcellular localization and downstream mechanism of XIST were investigated.Finally,a rescue experiment was carried out to verify that XIST regulates NLR family pyrin domain containing 3(NLRP3)/caspase-1-mediated RTEC pyroptosis through the microRNA-15-5p(miR-15b-5p)/Toll-like receptor 4(TLR4)axis.RESULTS XIST was highly expressed in the DN models.XIST silencing improved renal metabolism and biochemical indices and mitigated renal injury.The expression of inflammation and pyroptosis indicators was significantly increased in DN rats and HG-treated HK2 cells;cell viability was decreased and LDH activity was increased after HGtreatment. Silencing XIST inhibited RTEC pyroptosis by inhibiting NLRP3/caspase-1. Mechanistically,XIST sponged miR-15b-5p to regulate TLR4. Silencing XIST inhibited TLR4 by promotingmiR-15b-5p. miR-15b-5p inhibition or TLR4 overexpression averted the inhibitory effect ofsilencing XIST on HG-induced RTEC pyroptosis.CONCLUSIONSilencing XIST inhibits TLR4 by upregulating miR-15b-5p and ultimately inhibits renal injury inDN by inhibiting NLRP3/caspase-1-mediated RTEC pyroptosis.

The hepatitis C virus 5’ untranslated region gene amplified by rapid amplification of cDNA ends and its secondary structure

Objectives: To obtain very end full-length cDNA ofhepatitis C virus (HCV) 5’ untranslated region(5’UTR) and analyze its primary and secondarystructure.Methods: A patient infected genotype 2a HCV wasidentified by reverse transcription-nested polymerasechain reaction (RT-PCR) and restriction fragmentlength polymorphism (RFLP). Total RNA isolatedfrom the serum was used as template, and the cDNAof the 5’ untranslated region was amplified using rap-id amplification of cDNA ends (RACE). The frag-ments were recombinated by A-T clone strategy, andthe recombinants were confirmed by RFLP andPCR, and sequenced subsequently. Secondary struc-tures were analysed by RNAdraw.Results: Very end full-length cDNA of genotype 2aHCV 5’ UTR was obtained by RACE. In five clonesobtained, three contained full-length 5’UTR cDNA;A21G, G170A, T222C, T247C, C339T substitutionswere found as compared to HC-J6. Homological re-sults of HCV-1, HC-J6, HC-C2, HC-J8 were 93.6%-94.4%, 92.1%-93%, 98.8%-99.7%, 96.2%-96.5%, respectively; however, the substitutions didnot alter secondary structure. Two of 5 clones weredeletions of 53bp and 135bp at the 5’terminal ofHCV 5’UTR, respectively.Conclusions: RACE can be used to obtain the full-length cDNA of 2a genotype HCV 5’UTR. Genes de-leted at the 5’ terminal of HCV circulate in hepatitisC patients.

Identification of key long noncoding RNAs and their biological functions in hepatocellular carcinoma

Long noncoding RNAs(lncRNAs)are vital regulators in tumorigenesis and metastasis.However,the pathological role of lncRNAs in hepatocellular carcinoma(HCC)is still unclear.In this study,we filtered out three lncRNAs from The Cancer Genome Atlas(TCGA)data that were screened for basic expression and clinical research.We selected lncRNA-NEAT1 for further study to explore its function in HCC progression and its regulatory mechanism.We identified three differentially expressed lncRNAs(DElncRNAs)in tumor and adjacent normal tissues from the TCGA library using data mining methods:lncRNA-NEAT1,lncRNA-MAGI2-AS3 and lncRNA-HCG11.Their basic expression levels were detected by qPCR.Then,we selected lncRNA-NEAT1 as a potentially important lncRNA to verity its biological function and mechanism in HCC cell lines.lncRNA-NEAT1,lncRNA-MAGI2-AS3 and lncRNA-HCG11 were overexpressed in liver cancer tissues and cell lines.We found that silencing NEAT1 in vitro can inhibit the proliferation of HuH-7 and Li-7 cells,inhibit cell migration,and induce apoptosis as well as significantly increase the level of miR-16-5p.We also confirmed that miR-16-5p has a significant correlation with Bcl-2.When NEAT1 is silenced,the expression of Bcl-2 decreases.Inhibiting miR-16-5p can restore Bcl-2 to its original level.We conclude that miR-16-5p1/lncRNA NEAT1 plays a crucial role in regulating the delivery of Bcl-2 in HCC.Overall,the miR-16-5p/lncRNA-NEAT1/Bcl-2 signaling axis may be a promising target for HCC treatment.

Differentially expressed long noncoding RNAs and regulatory mechanism of LINC02407 in human gastric adenocarcinoma

BACKGROUND Long noncoding RNAs(lncRNAs)have been identified to play important roles in the development and progression of various tumors,including gastric cancer(GC).However,the molecular role of lncRNAs in GC progression remains unclear.AIM To investigate the differential expression of lncRNAs in human GC and elucidate the function and regulatory mechanism of LINC02407.METHODS The Cancer Genome Atlas database was used to investigate the involvement of lncRNAs in GC.Quantitative real-time polymerase chain reaction was used to estimate the relative expression level of LINC02407 in GC tissues and cells.Functional experiments including CCK8 assay,apoptosis assay,wound healing assay,and transwell assay were used to investigate the effect of LINC02407 on GC cells.Some microRNAs were predicted and verified via bioinformatics analysis and the luciferase reporter system.Predictive analysis and Western blot assay were used to analyze the expression of related proteins.RESULTS Many differentially expressed lncRNAs were identified in GC,and some of them including LINC02407 can affect the survival.LINC02407 was upregulated in tumor tissues compared with adjacent tissues.HGC-27 cells showed the highest LINC02407 expression and HaCaT cells exhibited the lowest expression.Different experiment groups were constructed using LINC02407 overexpressing plasmids and related siRNAs.The results of functional experiments showed that LINC02407 can promote the proliferation,migration,and invasion of GC cells but inhibit apoptosis.Luciferase reporter assay showed that hsa-miR-6845-5p and hsa-miR-4455 was downstream regulated by LINC02407.Western blot analysis showed that adhesion G protein-coupled receptor D1(ADGRD1)was regulated by the LINC02407-miR-6845-5p/miR-4455-ADGRD1 pathways.CONCLUSION LINC02407 plays a role in GC through the LINC02407-miR-6845-5p/miR-4455-ADGRD1 pathways,and thus,it may be an important oncogene and has potential value in GC diagnosis and treatment.

5-HTTLPR基因多态性与惊恐障碍发病风险关系的Meta分析

目的系统评价5-羟色胺转运体启动子区(5-HTTLPR)基因多态性与惊恐障碍(PD)发病风险的关系。方法利用PubMed、Embase、中国知网、维普网和万方数据知识服务平台等数据库检索5-HTTLPR基因多态性与PD发病风险关系的相关文献,根据异质性检验结果,采用固定效应模型分析总体人群及以种族划分为亚组人群5种基因遗传模型下5-HTTLPR基因多态性与PD发病风险的关系。结果共纳入合格文献11篇,包括PD组1431例,对照组2148例。在总体人群及以种族划分为亚组人群中,5种基因遗传模型下5-HTTLPR基因多态性与PD发病风险均无关(均P>0.05)。结论5-HTTLPR基因多态性可能与PD发病风险无关。

Genomic structure analysis of SNC6, a progesterone-receptor associated protein gene, and cloning and characterization of its 5＇-flanking region

Objective: To analyze the genomic structure of SNC6, a progesterone\|receptor associated protein gene and its regulatory elements in its 5'\|flanking region. Methods: Genomic sequence from GenBank database (accession number: Z98048) covering the whole SNC6 gene was used to analyze the genomic structure of SNC6 and design primers for PCR amplification of its 5'\|flanking region. A 1894 bp fragment of the 5'\|flanking region \{(-1814\} to +75) was cloned by PCR using genomic DNA from a healthy donor peripheral blood lymphocyte as template. This fragment, as well as 3 shorter derivative fragments (1423 bp, 632 bp and 416 bp, which correspond to -1344 to +75, -552 to +75 and -337 to +75 respectively), were subcloned into pGL2 series luciferase reporter vectors. These constructs were introduced into colorectal cancer cell line SW620 for transient expression of reporter gene and luciferase activities were measured. Results: The genomic structure analysis showed there are 12 exons for SNC6 gene, which spans 32017 bp (nt71529 to nt39513 in Z98048 sequence). All transfected SW620 cells with the above 5\|flanking region\|containing constructs showed luciferase activities. The highest luciferase activities were measured in transfected cells with vectors containing 1894 bp fragments, and the lowest luciferase activities were measured in transfected cells with vectors containing 416 bp fragments. Luciferase activities were higher in transfected cells with vectors containing 632 bp fragments than that in transfected cells with vectors containing 1423 bp fragments. Conclusion: The basic transcription\|promoting element (promoter) for SNC6 expression resides between 0 to -337, and two transcription\|enhancing elements (enhancer) resides between -337 to -552 and -1344 to -1814, whereas one transcription\|inhibiting element (silencer) exists between -552 to -1344.