Objectives] This study aimed to determine the optimal medium components and culture conditions for the induction and proliferation of Dioscorea nipponica Makino axillary buds in vitro .[Methods] The effects of basic m...Objectives] This study aimed to determine the optimal medium components and culture conditions for the induction and proliferation of Dioscorea nipponica Makino axillary buds in vitro .[Methods] The effects of basic medium (MS, WPM, B5 and N6), cytokinins type and concentration (0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L 6-BA; 0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L KT), auxins type and concentration (0, 0.25, 0.50, 0.75, 1.00 mg/L IBA, 2,4-D and NAA) on the induction of D. nipponica axillary buds were respectively measured and compared. Then, the effects of sugar source (30 g/L sucrose, fructose, white sugar, maltose and glucose) and light intensity (0, 800, 1 600, 2 400 and 3 200 lx) on the proliferation coefficient of D. nipponica axillary buds were evaluated.[Results] Using stem segments with leaf axils as the explants, the highest induction rate (90.8%) of axillary buds was achieved in MS supplemented with 2.5 mg/L 6-BA and 0.5 mg/L 2,4-D, and the fresh and dry weights of tissue culture seedlings in this medium were also the highest, up to 1.5 g and 160.9 mg, respectively. Then, the highest proliferation coefficient of was D. nipponica axillary buds noted when sucrose was used as the sugar source in medium, and the optimal light intensity for the proliferation of D. nipponica axillary buds was 2 400 lx.[Conclusions] The results provide an experimental evidence for rapid propagation of D. nipponica .展开更多
[Objective]This study aimed to investigate the major factors influencing the proliferation and growth of aseptic buds of Vietnam Mesona blumes,thus providing technical references for establishing rapid propagation sys...[Objective]This study aimed to investigate the major factors influencing the proliferation and growth of aseptic buds of Vietnam Mesona blumes,thus providing technical references for establishing rapid propagation system in vitro of Vietnam M.blumes.[Method]The effects of medium components(different basic medium,sucrose concentrations and cytokinins)and explant materials(different explants)on the proliferation of aseptic buds of Vietnam M.blumes were analyzed.[Result]Among MS,Miller and modified White basic medium,MS medium exhibited the best effect on proliferation of Vietnam M.blumes buds with the bud proliferation multiple of 8.53.Among different sucrose concentrations,20.0 g/L sucrose gave the highest bud proliferation multiple of 8.34,while 25.0 g/L sucrose led to the best growth status with the bud proliferation multiple of 7.05.The effect of CPPU on bud proliferation was higher than that of 6-BA and KT,with the bud proliferation multiple of 9.80.Among different explants,top stem exhibited the best effect on bud proliferation with the bud proliferation multiple of 9.35,resulting in robust seedlings.[Conclusion]The most suitable subculture medium for the proliferation and growth of aseptic buds of Vietnam M.blumes is MS+20.0-25.0 g/L sucrose+0.5 mg/L CPPU and the most suitable explant is top stem.展开更多
Thamnocalamus falconeri, Hook.f. ex Munro.,an important bamboo species belonging to the family Poaceae, locally known as Ringal, occurs in the hills of Uttarakhand, India. This species has been traditionally exploited...Thamnocalamus falconeri, Hook.f. ex Munro.,an important bamboo species belonging to the family Poaceae, locally known as Ringal, occurs in the hills of Uttarakhand, India. This species has been traditionally exploited by local communities to support their livelihoods.Increasing needs of the hill villages impose unsustainable pressure on natural stands of Ringal in the Uttarakhand hills and forests have been degraded. The long history of excessive cutting of Ringal from natural forests and the lack of replanting threaten villager livelihoods. Replanting is required to conserve the species. We propose a protocol for generation of planting material through axillary bud proliferation for multiplication and conservation of this species. We collected offsets/rhizomes from a natural stand of T. falconeri in the Chopta Mandal areas(Chamoli district, India). These were planted at sites of varied elevation and fresh single nodal segments were collected from them as explants. Different sterilization treatments were assessed to combat contamination. Among these, treatment of 0.1 %Hg Cl2 followed by 5 % Na OCl, proved best. Among two cytokinin treatments, viz. BAP and Kinetin, singly or in combination, BAP alone(5 mg L-1) proved superior and resulted in 100 % bud break. BAP-supplemented MS media yielded maximum vigorous shoot formation(90 %)and maximum number of shoots(8.9). Subculturing of shoots on the same medium with similar BAP treatment(5 mg L-1BAP) enabled continuous production of healthy shoots at similar frequency. Maximum rooting(100 %)was recorded on half-strength MS medium supplemented with 5 mg L-1IBA. Micropropagated plants were hardened and acclimatized in soil mixture(2:1:1) and then transplanted to field sites(Magra, Uttarakhand, 1,834 m).Eight to ten months after field transplantation we recorded100 % survival of transplanted material. This micropropagation protocol could be used successfully for raising a stock of genetically homogenous plant material in bulk for field plantations and for conservation of the species.展开更多
AIM:To compare the impact of ErbB2 on cell invasion and proliferation in cholangiocarcinoma(CCA) cell lines.METHODS:Level of endogenous ErbB2 expression in three CCA cell lines,namely HuCCA-1,KKU-100 and KKU-M213,was ...AIM:To compare the impact of ErbB2 on cell invasion and proliferation in cholangiocarcinoma(CCA) cell lines.METHODS:Level of endogenous ErbB2 expression in three CCA cell lines,namely HuCCA-1,KKU-100 and KKU-M213,was determined by real-time reversetranscriptase polymerase chain reaction.Two ErbB2 inhibitory methods,a small molecule ErbB2 kinase inhibitor(AG825) and siRNA,were used to disrupt ErbB2 function in the cell lines.CCA cell invasion,motility and proliferation under ErbB2-disrupted conditions were detected using Transwell and 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide assays.In addition,ErbB2 downstream effectors were investigated by Western blotting analysis.RESULTS:Suppression of ErbB2 activity,using a specific kinase inhibitor(AG825),reduced invasion,motility and proliferation of all three CCA cell lines.The ability of this drug to inhibit neoplastic properties(invasion,motility and proliferation) increased concomitantly with the level of ErbB2 expression.Similarly,knockdown of ErbB2 level by siRNA inhibited cell invasion and proliferation of KKU-M213,a high-ErbB2-expressing cell,better than those of the lower-ErbB2-expressing cells,HuCCA-1 and KKU-100.Thus,both inhibitory methods indicated that there is more ErbB2-dependency for malignancy of the high-ErbB2-expressing cell,KKU-M213,than for that of low-ErbB2-expressing ones.In addition,interrupting ErbB2 activity decreased phosphorylation of AKT and p70S6K,but not extracellular signal-regulated kinase 1/2,in the high-ErbB2-expressing CCA cell line.CONCLUSION:Our data indicated that high ErbB2 expression enhances CCA invasion,motility and proliferation via the AKT/p70S6K pathway,which suggests the possibility of targeting these molecules for CCA therapy.展开更多
Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferatio...Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferation. Here, the effects of the LRP16 gene expression on growth of MCF-7 human breast cancer cells and the mechanism were further studied by establishing two stably LRP16-inhibitory MCR-7 cell lines. Methods: Hairpin small interference RNA (siRNA) strategy, by which hairpin siRNA was released by U6 promoter and was mediated by pLPC-based retroviral vector, was adopted to knockdown endogenous LRP16 level in MCF-7 cells. And the hairpin siRNA against green fluorescence protein (GFP) was used as the negative control. The suppressant efficiency of the LRP16 gene expression was confirmed by Nothern blot. Cell proliferation assay and soft agar colony formation assay were used to determine the status of the cells proliferation. Cell cycle checkpoints including cyclin E and cyclin D1 were examined by Western blot. Results: The results from cell proliferation assays suggested that down-regulation of LRP16 gene expression is capable of inhibiting MCF-7 breast cancer cell growth and down-regulation of the LRP16 gene expression is able to inhibit anchorage-independent growth of breast cancer cells in soft agar. We also demonstrated that cyclin E and cyclin D1 proteins were much lower in the LRP16-inhibitory cells than in the control cells. Conclusion: These data suggest that LRP16 gene play an important role in MCF-7 cells proliferation by regulating the pathway of the G1/S transition and may function as an important modulator in regulating the process of tumorigenesis in human breast.展开更多
Objective:To study whether the infection of Schistosomiasis japanicum(S.japanicum) is related to enhanced proliferation and migration of cancer cells,and the molecular mechanism pertains to cancer cell metastasis in h...Objective:To study whether the infection of Schistosomiasis japanicum(S.japanicum) is related to enhanced proliferation and migration of cancer cells,and the molecular mechanism pertains to cancer cell metastasis in human host.Methods:The gene of S.japanicum glutathione transferase(sjGST) cloned from 5.japanicum was expressed,purified and applied in a series of assays to explore the effect of sjGST on proliferation and migration of MDA-MB-435S,and the expression of MMP2 and MMP9.Immunofluorescence assay for the binding of sjGST to MDA-MB-435S was also carried out.Results:Results showed that sjGST enhanced proliferation and migration in human breast cancer cell MDA-MB-435S signifycantly at 50-200 nM,but did not enhance them in human lung cancer cell A549.Immunofluorescence assay for the binding of sjGST to MDA-MB-435S and A549 showed that GST was readily hound to the breast cancer cells,but showed almost no binding to human lung cancer cells.The assays for gelatinase activity showed that both MMP2 and MMP9 activities were increased significantly in the presence of sjGST(50-200 nM) in MDA-MB-435S, but they were not significant in A549.Conclusions:Our current results show strongly that S. japanicum GST binds to MDA-MB-435S probably via its i'eceptor,and enhances proliferation and migration of the cancer cells by up-regulatory expression of MMP2 and MMP9.展开更多
Objective To investigate the effect of exogenous S100A13 gene overexpression on the proliferation of human thyroid cancer cell line TT.Methods The recombinant ORF of S100A13 tagged with six histidines at the 5' en...Objective To investigate the effect of exogenous S100A13 gene overexpression on the proliferation of human thyroid cancer cell line TT.Methods The recombinant ORF of S100A13 tagged with six histidines at the 5' end was subcloned into the pcDNA3.2/V5/GW/D-TOPO vector and sequenced.The eukaryotic expression plasmid pcDNA3.2/V5 /GW/D-S100A13 and empty vector pcDNA3.2/V5/GW/D were transfected into TT cells.The positive clones were selected by G418.The expressions of S100A13 mRNA and protein were detected by real time reverse transcription-polymerase chain reaction(RT-PCR) and Western blot.The effect of S100A13 on cell proliferation and cell cycle was evaluated by cell growth curve,MTT colorimetric assay and flow cytometry.Results S100A13 gene tagged with six histidines at the 5 ' end was confirmed to be inserted into the pcDNA3.2/V5/GW/D vector correctly.TT-S100A13-V5 cells,which over-expressed S100A13,were constructed successfully.TT-S100A13-V5 cells grew much faster than TT-V5 and TT cells(P <0.001).The proportions of both S and G2/M phase cells were significantly higher in TT-S100A13-V5 cells than those in TT-V5 and TT cells(P <0.001).Conclusion The eukaryotic expression vector containing human S100A13 gene has been successfully constructed,which highly expresses S100A13 in TT cells.Exogenous S100A13 gene overexpression accelerates TT cell proliferation and drives the cell cycle progression of TT cells from G0/G1 phase to S and G2/M phases.展开更多
AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells ...AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells in vitro and on mouse implanted S-180tumors in vivo.METHODS The polysaccharide extracts were isolated from the fruit bodies of FV, LE and AB by the methods of hot-water extraction, Sevag’sremoval of proteins, ethanol precipitation,trypsin digestion and ethanol fractionalprecipitation. Human hepatoma SMMC-7721 cells were treated with 50 mg/L Polysaccharide extracts, and the mitosis index, mitochondria activity and cell proliferation were detected at different times in both control and experimental groups. The mice with S-180 implanted tumors were injected with the polysaccharide extracts at 24 mg/ kg body weight for 9 d and the tumorweight was measured on the 15th day.RESULTS The mitosis index of hepatoma cells in vitro could be significantly decreased by treatment with the polysaccharide extracts fromthe three kinds of edible fungi (P < 0 .005 ). Thecell numbers and mitochondria activity of SMMC7721 cells treated with polysaccharide extracts were lower than those in control groups (P <0.005). The inhibition rates of polysaccharide extracts against implanted S-180 tumors in mice were 52.8%, 56.6% and 51 .9% respectivelycompared with that in c0ntrol gr0ups.CONCLUSI0N The POIysaccharide extractsfrom the three kinds of edible fungi could inhibitnot only the Cultured malignant cells in vitfO butalso impIanted Sl80 tum0r i0 vivo.展开更多
The increasing amounts of artificial marine substrates, in many parts of the world have been proposed as a potential driver of Aurelia spp. blooms, on account of providing extra habitats for the settlement and the pro...The increasing amounts of artificial marine substrates, in many parts of the world have been proposed as a potential driver of Aurelia spp. blooms, on account of providing extra habitats for the settlement and the proliferation of the benthic stage(polyps). Previous experiments have mainly focused on the substrate choices of Aurelia spp. planulae. However, substrate preferences for the proliferation and immigration of polyps have not been reported. We monitored the propagation and immigration of Aurelia aurita(s. l.) polyps on two natural and nine artificial substrates at constant temperature(20±0.5°C) and salinity(30±0.5) in beakers and a glass aquarium in the laboratory, respectively. The results showed that, among artificial substrates, the highest number for polyp proliferation and immigration was found on nets, rigid polyvinyl chloride plates(RPVC), and wood. The lowest density of polyps was present on iron plates. Among natural substrates, the asexual reproduction rate of polyps on Patinopecten yessoensis(Jay, 1857) shells was significantly higher than Azumapecten farreri(Jones & Preston, 1904). On the account of the distinction in the roughness, chemical properties and biofilms of these material surfaces, bare artificial or natural substrates discriminatively affect the proliferation and the immigration of Aurelia spp. polyps at laboratory. These observations suggest that, even in the natural environment, different materials and texture may influence the composition and the abundance of the fouling communities and the assemblages of polyps and, indirectly, have effects on the amounts of released medusae.展开更多
[Objectives]The paper was to study the tissue culture nursery technology of Curcuma alismatifolia‘Kimono Rose’.[Methods]By sterilizing and inoculating explants derived from disparate regions of C.alismatifolia,we id...[Objectives]The paper was to study the tissue culture nursery technology of Curcuma alismatifolia‘Kimono Rose’.[Methods]By sterilizing and inoculating explants derived from disparate regions of C.alismatifolia,we identified the most optimal explants and optimized the culture conditions for cluster buds induction and proliferation.This was achieved by incorporating MS medium with varying concentrations of 6-BA and NAA,thereby establishing a foundation for the large-scale production of C.alismatifolia tissue culture seedlings.[Results]The optimal explant for C.alismatifolia was identified as a lateral bud.The most effective cluster buds induction medium was determined to be MS+6-BA 5 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.The optimal cluster buds proliferation medium was found to be MS+6-BA 3 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.[Conclusions]The findings of this study can provide a foundation for the enhancement of the industrialized breeding system of tissue culture propagation of C.alismatifolia.展开更多
Marek’s disease(MD),a highly cell-associated and contagious disease of chickens caused by Marek’s disease virus(MDV)can result in neural lesions,immunosuppression and neoplasia in chicken.The Meq gene is an importan...Marek’s disease(MD),a highly cell-associated and contagious disease of chickens caused by Marek’s disease virus(MDV)can result in neural lesions,immunosuppression and neoplasia in chicken.The Meq gene is an important oncogene in the MDV genome,and it is expressed highly in MD tumor tissues and MD T-lymphoblastoid cell lines.An experiment was conducted to elucidate the role of Meq in MD tumor transformation.RNA interference technology was used to block its expression,and then analyzed the biological effects of Meq knockdown on the MD tumor cell line MSB1.A small interfering RNA with an interference efficiency of 70%(P<0.01)was transfected into MSB1 cells to knock down the expression of Meq gene.The cell proliferation,cycle and apoptosis were detected post-Meq knockdown.The results showed that MSB1 cell proliferation was downregulated remarkably at 48 h(P<0.01),60 h(P<0.05)and 72 h(P<0.01)post-Meq knockdown.The cell cycle was unaffected(P>0.05).B-cell lymphoma 2 gene(BCL2)was anti-apoptotic and caspase-6 was the effector in the apoptosis pathway.The activity of caspase-6 was upregulated(P<0.05)significantly and BCL2 gene expression was downregulated(P<0.05)significantly post-Meq knockdown,suggesting cell apoptosis might be induced.MSB1 cell migration did not exhibit any obvious change(P>0.05)post-Meq knockdown,but the expression of two genes(matrix metalloproteinase 2(MMP2)and MMP9)that are correlated closely to cell invasion was downregulated(P<0.05)remarkably post-Meq knockdown.The Meq knockdown might affect the main features of tumorous cells,including proliferation,apoptosis,and invasion,suggesting that the Meq gene might play a crucial role in interfering with lymphomatous cell transformation.展开更多
Torin 1 is an ATP-competitive TOR inhibitor which inhibits the signaling of TOR and S6K kinase in mammals and plants.The objective of this research is to determine the effect of Torin 1 in a relatively simple and homo...Torin 1 is an ATP-competitive TOR inhibitor which inhibits the signaling of TOR and S6K kinase in mammals and plants.The objective of this research is to determine the effect of Torin 1 in a relatively simple and homogeneous plant system such as the NT-1 tobacco suspension cell cultures.Cultures of NT-1 cells were tested with 5,50,150 and 250 nM of Torin 1.During kinetics growth of NT-1 tobacco suspension cell cultures,150 and 250 nM Torin 1 inhibits the early growth and later enhanced the cellular proliferation during exponential growth by means of an increased expression of E2F1 and cyclin B.Furthermore,Torin 1 stimulates the growth of NT-1 cells during log phase with small shaped cell,characteristic of tobacco suspension cell cultures with high mitotic activity.展开更多
Neurons are believed to be non-proliferating cells.However,neuronal stem cells are still present in certain areas of the adult brain,although their proliferation diminishes with age.Just as with other cells,their prol...Neurons are believed to be non-proliferating cells.However,neuronal stem cells are still present in certain areas of the adult brain,although their proliferation diminishes with age.Just as with other cells,their proliferation and differentiation are modulated by various mechanisms.These mechanisms are foundational to the strategies developed to induce neuronal proliferation and differentiation,with potential therapeutic applications for neurodegenerative diseases.The most common among these diseases are Parkinson's disease and Alzheimer's disease,associated with the formation ofβ-amyloid(Aβ)aggregates which cause a reduction in the number of neurons.Compounds such as LiCl,4-aminothiazoles,Pregnenolone,ACEA,harmine,D2AAK1,methyl 3,4-dihydroxybenzoate,and shikonin may induce neuronal proliferation/differentiation through the activation of pathways:MAPK ERK,PI3K/AKT,NFκB,Wnt,BDNF,and NPAS3.Moreover,combinations of these compounds can potentially transform somatic cells into neurons.This transformation process involves the activation of neuron-specific transcription factors such as NEUROD1,NGN2,ASCL1,and SOX2,which subsequently leads to the transcription of downstream genes,culminating in the transformation of somatic cells into neurons.Neurodegenerative diseases are not the only conditions where inducing neuronal proliferation could be beneficial.Consequently,the impact of pro-proliferative compounds on neurons has also been researched in mouse models of Alzheimer's disease.展开更多
Cohesin is an evolutionary conserved complex that controls chromosome segregation during mitosis. Here we show that, in response to DNA damage, Saccharomyces cerevisiae Scc3, one of the major regulatory subunits of th...Cohesin is an evolutionary conserved complex that controls chromosome segregation during mitosis. Here we show that, in response to DNA damage, Saccharomyces cerevisiae Scc3, one of the major regulatory subunits of the Smc1-Smc3-Scc1 cohesin ring, is phosphorylated on S/T-Q residues. This event depended on the Mec1 checkpoint kinase as well as on cell cycle arrest triggered by the DNA damage checkpoint network. This phosphorylation event also took place during mitosis of an unperturbed cell cycle. The present finding that S. cerevisiae Scc3 is phosphorylated during mitosis represents a potentially important new regulatory step in cohesin’s mitotic functions.展开更多
Severe growth abnormalities including shoot stunting, leaf blade reduction and flower bud failure of Brussels sprout were observed in Poland. The presence of phytoplasma in diseased as well as in healthy looking plant...Severe growth abnormalities including shoot stunting, leaf blade reduction and flower bud failure of Brussels sprout were observed in Poland. The presence of phytoplasma in diseased as well as in healthy looking plants, was demonstrated by nested polymerase chain reaction assay employing phytoplasma universal rRNA primer pairs-P1/P7 followed by R16F2n/R16R2. Products of PCR primed by R 16F2n/R 16R2 primer pair from naturally infected Brussels sprouts were sequenced. Comparison of the obtained 16S rDNAs revealed high nucleotide sequence identity between analyzed phytoplasma isolates (99.8%-100%). They were also nearly identical with the sequences of other phytoplasmas isolates from sub-group 16SrI-B, and they were classified as members of "Candidatus Phytoplasma asteris".展开更多
目的探讨人参皂苷20(S)-Rg3抑制卵巢癌细胞增殖和迁移的机制。方法在西安交通大学第一附属医院收集卵巢癌组织19例,正常卵巢组织18例,采用real-time PCR检测lncRNA KRT18P55的表达。将卵巢癌细胞SKOV3和3AO分别分为2组:阴性对照组和20(S...目的探讨人参皂苷20(S)-Rg3抑制卵巢癌细胞增殖和迁移的机制。方法在西安交通大学第一附属医院收集卵巢癌组织19例,正常卵巢组织18例,采用real-time PCR检测lncRNA KRT18P55的表达。将卵巢癌细胞SKOV3和3AO分别分为2组:阴性对照组和20(S)-Rg3组,通过real-time PCR检测KRT18P55的表达水平,CCK-8和平板克隆实验检测SKOV3和3AO细胞增殖能力,Transwell小室法检测SKOV3和3AO细胞迁移能力。将KRT18P55 siRNA分别转染SKOV3和3AO细胞,通过real-time PCR检测KRT18P55的表达水平,CCK-8和平板克隆实验检测SKOV3和3AO细胞增殖能力,Transwell小室法检测SKOV3和3AO细胞迁移能力。癌症基因组学门户网站(cBioPortal for Cancer Genomics,cBioPortal)提取KRT18P55共表达基因数据;基因功能分析数据库(Gene Annotation and Analysis Resource,Metascape)对KRT18P55及其显著相关基因进行功能富集分析;基因表达谱交互式分析(Gene Expression Profiling Interactive Analysis,GEPIA)、阿拉巴马大学癌症数据分析门户(University of Alabama at Birmingham Cancer Data Analysis Portal,UALCAN)在线数据库对筛选的KRT18P55显著相关基因进行表达分析。结果人卵巢癌组织中的KRT18P55表达水平显著高于正常卵巢组织,经20(S)-Rg3处理的卵巢癌细胞SKOV3和3AO的增殖和迁移能力均下降,KRT18P55的表达水平显著降低,差异均具有统计学意义(P<0.05)。敲低KRT18P55后,卵巢癌细胞的增殖和迁移能力下降(P<0.05)。基因本体(Gene Ontology,GO)功能富集分析显示,KRT18P55可能参与中间丝细胞骨架组成、外源性凋亡信号通路等。GEPIA及UALCAN数据库的表达分析显示,KRT18P55相关性最强的两个基因KRT18、KRT8的mRNA及蛋白水平在卵巢癌中均较正常对照显著增高(P<0.05)。结论人参皂苷20(S)-Rg3可能通过降低KRT18P55的表达水平抑制卵巢癌细胞的增殖与迁移,进而抑制卵巢癌的进展。展开更多
基金Supported by University-enterprise Cooperation Project of Yanbian University([2006]3)
文摘Objectives] This study aimed to determine the optimal medium components and culture conditions for the induction and proliferation of Dioscorea nipponica Makino axillary buds in vitro .[Methods] The effects of basic medium (MS, WPM, B5 and N6), cytokinins type and concentration (0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L 6-BA; 0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L KT), auxins type and concentration (0, 0.25, 0.50, 0.75, 1.00 mg/L IBA, 2,4-D and NAA) on the induction of D. nipponica axillary buds were respectively measured and compared. Then, the effects of sugar source (30 g/L sucrose, fructose, white sugar, maltose and glucose) and light intensity (0, 800, 1 600, 2 400 and 3 200 lx) on the proliferation coefficient of D. nipponica axillary buds were evaluated.[Results] Using stem segments with leaf axils as the explants, the highest induction rate (90.8%) of axillary buds was achieved in MS supplemented with 2.5 mg/L 6-BA and 0.5 mg/L 2,4-D, and the fresh and dry weights of tissue culture seedlings in this medium were also the highest, up to 1.5 g and 160.9 mg, respectively. Then, the highest proliferation coefficient of was D. nipponica axillary buds noted when sucrose was used as the sugar source in medium, and the optimal light intensity for the proliferation of D. nipponica axillary buds was 2 400 lx.[Conclusions] The results provide an experimental evidence for rapid propagation of D. nipponica .
基金Supported by Scientific Research and Technological Development Project of Wuzhou City,Guangxi Zhuang Autonomous Region(200901024)
文摘[Objective]This study aimed to investigate the major factors influencing the proliferation and growth of aseptic buds of Vietnam Mesona blumes,thus providing technical references for establishing rapid propagation system in vitro of Vietnam M.blumes.[Method]The effects of medium components(different basic medium,sucrose concentrations and cytokinins)and explant materials(different explants)on the proliferation of aseptic buds of Vietnam M.blumes were analyzed.[Result]Among MS,Miller and modified White basic medium,MS medium exhibited the best effect on proliferation of Vietnam M.blumes buds with the bud proliferation multiple of 8.53.Among different sucrose concentrations,20.0 g/L sucrose gave the highest bud proliferation multiple of 8.34,while 25.0 g/L sucrose led to the best growth status with the bud proliferation multiple of 7.05.The effect of CPPU on bud proliferation was higher than that of 6-BA and KT,with the bud proliferation multiple of 9.80.Among different explants,top stem exhibited the best effect on bud proliferation with the bud proliferation multiple of 9.35,resulting in robust seedlings.[Conclusion]The most suitable subculture medium for the proliferation and growth of aseptic buds of Vietnam M.blumes is MS+20.0-25.0 g/L sucrose+0.5 mg/L CPPU and the most suitable explant is top stem.
文摘Thamnocalamus falconeri, Hook.f. ex Munro.,an important bamboo species belonging to the family Poaceae, locally known as Ringal, occurs in the hills of Uttarakhand, India. This species has been traditionally exploited by local communities to support their livelihoods.Increasing needs of the hill villages impose unsustainable pressure on natural stands of Ringal in the Uttarakhand hills and forests have been degraded. The long history of excessive cutting of Ringal from natural forests and the lack of replanting threaten villager livelihoods. Replanting is required to conserve the species. We propose a protocol for generation of planting material through axillary bud proliferation for multiplication and conservation of this species. We collected offsets/rhizomes from a natural stand of T. falconeri in the Chopta Mandal areas(Chamoli district, India). These were planted at sites of varied elevation and fresh single nodal segments were collected from them as explants. Different sterilization treatments were assessed to combat contamination. Among these, treatment of 0.1 %Hg Cl2 followed by 5 % Na OCl, proved best. Among two cytokinin treatments, viz. BAP and Kinetin, singly or in combination, BAP alone(5 mg L-1) proved superior and resulted in 100 % bud break. BAP-supplemented MS media yielded maximum vigorous shoot formation(90 %)and maximum number of shoots(8.9). Subculturing of shoots on the same medium with similar BAP treatment(5 mg L-1BAP) enabled continuous production of healthy shoots at similar frequency. Maximum rooting(100 %)was recorded on half-strength MS medium supplemented with 5 mg L-1IBA. Micropropagated plants were hardened and acclimatized in soil mixture(2:1:1) and then transplanted to field sites(Magra, Uttarakhand, 1,834 m).Eight to ten months after field transplantation we recorded100 % survival of transplanted material. This micropropagation protocol could be used successfully for raising a stock of genetically homogenous plant material in bulk for field plantations and for conservation of the species.
基金Supported by A grant from Mahidol University,Thailand (to Suthiphongchai T)a scholarship from the Royal Golden Jubilee PhD Program,the Thailand Research Fund (to Treekit-karnmongkol W)
文摘AIM:To compare the impact of ErbB2 on cell invasion and proliferation in cholangiocarcinoma(CCA) cell lines.METHODS:Level of endogenous ErbB2 expression in three CCA cell lines,namely HuCCA-1,KKU-100 and KKU-M213,was determined by real-time reversetranscriptase polymerase chain reaction.Two ErbB2 inhibitory methods,a small molecule ErbB2 kinase inhibitor(AG825) and siRNA,were used to disrupt ErbB2 function in the cell lines.CCA cell invasion,motility and proliferation under ErbB2-disrupted conditions were detected using Transwell and 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide assays.In addition,ErbB2 downstream effectors were investigated by Western blotting analysis.RESULTS:Suppression of ErbB2 activity,using a specific kinase inhibitor(AG825),reduced invasion,motility and proliferation of all three CCA cell lines.The ability of this drug to inhibit neoplastic properties(invasion,motility and proliferation) increased concomitantly with the level of ErbB2 expression.Similarly,knockdown of ErbB2 level by siRNA inhibited cell invasion and proliferation of KKU-M213,a high-ErbB2-expressing cell,better than those of the lower-ErbB2-expressing cells,HuCCA-1 and KKU-100.Thus,both inhibitory methods indicated that there is more ErbB2-dependency for malignancy of the high-ErbB2-expressing cell,KKU-M213,than for that of low-ErbB2-expressing ones.In addition,interrupting ErbB2 activity decreased phosphorylation of AKT and p70S6K,but not extracellular signal-regulated kinase 1/2,in the high-ErbB2-expressing CCA cell line.CONCLUSION:Our data indicated that high ErbB2 expression enhances CCA invasion,motility and proliferation via the AKT/p70S6K pathway,which suggests the possibility of targeting these molecules for CCA therapy.
基金This work was supported by NationalNatural Science Foundation of China (No. 30200095).
文摘Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferation. Here, the effects of the LRP16 gene expression on growth of MCF-7 human breast cancer cells and the mechanism were further studied by establishing two stably LRP16-inhibitory MCR-7 cell lines. Methods: Hairpin small interference RNA (siRNA) strategy, by which hairpin siRNA was released by U6 promoter and was mediated by pLPC-based retroviral vector, was adopted to knockdown endogenous LRP16 level in MCF-7 cells. And the hairpin siRNA against green fluorescence protein (GFP) was used as the negative control. The suppressant efficiency of the LRP16 gene expression was confirmed by Nothern blot. Cell proliferation assay and soft agar colony formation assay were used to determine the status of the cells proliferation. Cell cycle checkpoints including cyclin E and cyclin D1 were examined by Western blot. Results: The results from cell proliferation assays suggested that down-regulation of LRP16 gene expression is capable of inhibiting MCF-7 breast cancer cell growth and down-regulation of the LRP16 gene expression is able to inhibit anchorage-independent growth of breast cancer cells in soft agar. We also demonstrated that cyclin E and cyclin D1 proteins were much lower in the LRP16-inhibitory cells than in the control cells. Conclusion: These data suggest that LRP16 gene play an important role in MCF-7 cells proliferation by regulating the pathway of the G1/S transition and may function as an important modulator in regulating the process of tumorigenesis in human breast.
基金Supported by grants from National Science Council(NSC98-2314-B-110-001-MY3)
文摘Objective:To study whether the infection of Schistosomiasis japanicum(S.japanicum) is related to enhanced proliferation and migration of cancer cells,and the molecular mechanism pertains to cancer cell metastasis in human host.Methods:The gene of S.japanicum glutathione transferase(sjGST) cloned from 5.japanicum was expressed,purified and applied in a series of assays to explore the effect of sjGST on proliferation and migration of MDA-MB-435S,and the expression of MMP2 and MMP9.Immunofluorescence assay for the binding of sjGST to MDA-MB-435S was also carried out.Results:Results showed that sjGST enhanced proliferation and migration in human breast cancer cell MDA-MB-435S signifycantly at 50-200 nM,but did not enhance them in human lung cancer cell A549.Immunofluorescence assay for the binding of sjGST to MDA-MB-435S and A549 showed that GST was readily hound to the breast cancer cells,but showed almost no binding to human lung cancer cells.The assays for gelatinase activity showed that both MMP2 and MMP9 activities were increased significantly in the presence of sjGST(50-200 nM) in MDA-MB-435S, but they were not significant in A549.Conclusions:Our current results show strongly that S. japanicum GST binds to MDA-MB-435S probably via its i'eceptor,and enhances proliferation and migration of the cancer cells by up-regulatory expression of MMP2 and MMP9.
基金supported by the Natural Science Fundof Hunan Province(No.06jj5046,No.05jj30039)
文摘Objective To investigate the effect of exogenous S100A13 gene overexpression on the proliferation of human thyroid cancer cell line TT.Methods The recombinant ORF of S100A13 tagged with six histidines at the 5' end was subcloned into the pcDNA3.2/V5/GW/D-TOPO vector and sequenced.The eukaryotic expression plasmid pcDNA3.2/V5 /GW/D-S100A13 and empty vector pcDNA3.2/V5/GW/D were transfected into TT cells.The positive clones were selected by G418.The expressions of S100A13 mRNA and protein were detected by real time reverse transcription-polymerase chain reaction(RT-PCR) and Western blot.The effect of S100A13 on cell proliferation and cell cycle was evaluated by cell growth curve,MTT colorimetric assay and flow cytometry.Results S100A13 gene tagged with six histidines at the 5 ' end was confirmed to be inserted into the pcDNA3.2/V5/GW/D vector correctly.TT-S100A13-V5 cells,which over-expressed S100A13,were constructed successfully.TT-S100A13-V5 cells grew much faster than TT-V5 and TT cells(P <0.001).The proportions of both S and G2/M phase cells were significantly higher in TT-S100A13-V5 cells than those in TT-V5 and TT cells(P <0.001).Conclusion The eukaryotic expression vector containing human S100A13 gene has been successfully constructed,which highly expresses S100A13 in TT cells.Exogenous S100A13 gene overexpression accelerates TT cell proliferation and drives the cell cycle progression of TT cells from G0/G1 phase to S and G2/M phases.
文摘AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells in vitro and on mouse implanted S-180tumors in vivo.METHODS The polysaccharide extracts were isolated from the fruit bodies of FV, LE and AB by the methods of hot-water extraction, Sevag’sremoval of proteins, ethanol precipitation,trypsin digestion and ethanol fractionalprecipitation. Human hepatoma SMMC-7721 cells were treated with 50 mg/L Polysaccharide extracts, and the mitosis index, mitochondria activity and cell proliferation were detected at different times in both control and experimental groups. The mice with S-180 implanted tumors were injected with the polysaccharide extracts at 24 mg/ kg body weight for 9 d and the tumorweight was measured on the 15th day.RESULTS The mitosis index of hepatoma cells in vitro could be significantly decreased by treatment with the polysaccharide extracts fromthe three kinds of edible fungi (P < 0 .005 ). Thecell numbers and mitochondria activity of SMMC7721 cells treated with polysaccharide extracts were lower than those in control groups (P <0.005). The inhibition rates of polysaccharide extracts against implanted S-180 tumors in mice were 52.8%, 56.6% and 51 .9% respectivelycompared with that in c0ntrol gr0ups.CONCLUSI0N The POIysaccharide extractsfrom the three kinds of edible fungi could inhibitnot only the Cultured malignant cells in vitfO butalso impIanted Sl80 tum0r i0 vivo.
基金Supported by the State Key Program of National Natural Science of China(No.41230963)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDA11020305)+1 种基金the Shandong Joint Fund for Marine Ecology and Environmental Sciences(No.U1406403)the National Natural Science Foundation of China(No.41506144)
文摘The increasing amounts of artificial marine substrates, in many parts of the world have been proposed as a potential driver of Aurelia spp. blooms, on account of providing extra habitats for the settlement and the proliferation of the benthic stage(polyps). Previous experiments have mainly focused on the substrate choices of Aurelia spp. planulae. However, substrate preferences for the proliferation and immigration of polyps have not been reported. We monitored the propagation and immigration of Aurelia aurita(s. l.) polyps on two natural and nine artificial substrates at constant temperature(20±0.5°C) and salinity(30±0.5) in beakers and a glass aquarium in the laboratory, respectively. The results showed that, among artificial substrates, the highest number for polyp proliferation and immigration was found on nets, rigid polyvinyl chloride plates(RPVC), and wood. The lowest density of polyps was present on iron plates. Among natural substrates, the asexual reproduction rate of polyps on Patinopecten yessoensis(Jay, 1857) shells was significantly higher than Azumapecten farreri(Jones & Preston, 1904). On the account of the distinction in the roughness, chemical properties and biofilms of these material surfaces, bare artificial or natural substrates discriminatively affect the proliferation and the immigration of Aurelia spp. polyps at laboratory. These observations suggest that, even in the natural environment, different materials and texture may influence the composition and the abundance of the fouling communities and the assemblages of polyps and, indirectly, have effects on the amounts of released medusae.
基金Supported by Special Project of Public-interest Scientific Institutions of Fujian Province(2021R1011003).
文摘[Objectives]The paper was to study the tissue culture nursery technology of Curcuma alismatifolia‘Kimono Rose’.[Methods]By sterilizing and inoculating explants derived from disparate regions of C.alismatifolia,we identified the most optimal explants and optimized the culture conditions for cluster buds induction and proliferation.This was achieved by incorporating MS medium with varying concentrations of 6-BA and NAA,thereby establishing a foundation for the large-scale production of C.alismatifolia tissue culture seedlings.[Results]The optimal explant for C.alismatifolia was identified as a lateral bud.The most effective cluster buds induction medium was determined to be MS+6-BA 5 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.The optimal cluster buds proliferation medium was found to be MS+6-BA 3 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.[Conclusions]The findings of this study can provide a foundation for the enhancement of the industrialized breeding system of tissue culture propagation of C.alismatifolia.
基金The work was supported in part by the National Natural Science Foundation of China(31320103905 and U1901206)the Young Scientist Supporting Project,Program for Changjiang Scholars and Innovative Research Team in University(IRT_15R62)+6 种基金the earmarked fund for China Agriculture Research Systems(CARS-41)the National High Technology Development Plan of China(2013AA102501)the Farm Animals Germplasm Resource Bankthe Beijing Key Laboratory for Animal Genetic Improvement,the University Research Project of Anhui Province,China(KJ2020A0081)the Anhui Provincial Natural Science Foundation,China(2008085QC140)the Foundation of Anhui Science and Technology University,China(DKYJ201901)the Innovation Funds for Undergraduate Students of Anhui Province,China(S201910879019,S202010879109,and S202010879120).
文摘Marek’s disease(MD),a highly cell-associated and contagious disease of chickens caused by Marek’s disease virus(MDV)can result in neural lesions,immunosuppression and neoplasia in chicken.The Meq gene is an important oncogene in the MDV genome,and it is expressed highly in MD tumor tissues and MD T-lymphoblastoid cell lines.An experiment was conducted to elucidate the role of Meq in MD tumor transformation.RNA interference technology was used to block its expression,and then analyzed the biological effects of Meq knockdown on the MD tumor cell line MSB1.A small interfering RNA with an interference efficiency of 70%(P<0.01)was transfected into MSB1 cells to knock down the expression of Meq gene.The cell proliferation,cycle and apoptosis were detected post-Meq knockdown.The results showed that MSB1 cell proliferation was downregulated remarkably at 48 h(P<0.01),60 h(P<0.05)and 72 h(P<0.01)post-Meq knockdown.The cell cycle was unaffected(P>0.05).B-cell lymphoma 2 gene(BCL2)was anti-apoptotic and caspase-6 was the effector in the apoptosis pathway.The activity of caspase-6 was upregulated(P<0.05)significantly and BCL2 gene expression was downregulated(P<0.05)significantly post-Meq knockdown,suggesting cell apoptosis might be induced.MSB1 cell migration did not exhibit any obvious change(P>0.05)post-Meq knockdown,but the expression of two genes(matrix metalloproteinase 2(MMP2)and MMP9)that are correlated closely to cell invasion was downregulated(P<0.05)remarkably post-Meq knockdown.The Meq knockdown might affect the main features of tumorous cells,including proliferation,apoptosis,and invasion,suggesting that the Meq gene might play a crucial role in interfering with lymphomatous cell transformation.
文摘Torin 1 is an ATP-competitive TOR inhibitor which inhibits the signaling of TOR and S6K kinase in mammals and plants.The objective of this research is to determine the effect of Torin 1 in a relatively simple and homogeneous plant system such as the NT-1 tobacco suspension cell cultures.Cultures of NT-1 cells were tested with 5,50,150 and 250 nM of Torin 1.During kinetics growth of NT-1 tobacco suspension cell cultures,150 and 250 nM Torin 1 inhibits the early growth and later enhanced the cellular proliferation during exponential growth by means of an increased expression of E2F1 and cyclin B.Furthermore,Torin 1 stimulates the growth of NT-1 cells during log phase with small shaped cell,characteristic of tobacco suspension cell cultures with high mitotic activity.
基金This research was funded under OPUS grant from National Science Center(NCN,Poland),grant number UMO-2021/43/B/NZ7/01732(to Agnieszka A.Kaczor).Figures were created using BioRender.
文摘Neurons are believed to be non-proliferating cells.However,neuronal stem cells are still present in certain areas of the adult brain,although their proliferation diminishes with age.Just as with other cells,their proliferation and differentiation are modulated by various mechanisms.These mechanisms are foundational to the strategies developed to induce neuronal proliferation and differentiation,with potential therapeutic applications for neurodegenerative diseases.The most common among these diseases are Parkinson's disease and Alzheimer's disease,associated with the formation ofβ-amyloid(Aβ)aggregates which cause a reduction in the number of neurons.Compounds such as LiCl,4-aminothiazoles,Pregnenolone,ACEA,harmine,D2AAK1,methyl 3,4-dihydroxybenzoate,and shikonin may induce neuronal proliferation/differentiation through the activation of pathways:MAPK ERK,PI3K/AKT,NFκB,Wnt,BDNF,and NPAS3.Moreover,combinations of these compounds can potentially transform somatic cells into neurons.This transformation process involves the activation of neuron-specific transcription factors such as NEUROD1,NGN2,ASCL1,and SOX2,which subsequently leads to the transcription of downstream genes,culminating in the transformation of somatic cells into neurons.Neurodegenerative diseases are not the only conditions where inducing neuronal proliferation could be beneficial.Consequently,the impact of pro-proliferative compounds on neurons has also been researched in mouse models of Alzheimer's disease.
文摘Cohesin is an evolutionary conserved complex that controls chromosome segregation during mitosis. Here we show that, in response to DNA damage, Saccharomyces cerevisiae Scc3, one of the major regulatory subunits of the Smc1-Smc3-Scc1 cohesin ring, is phosphorylated on S/T-Q residues. This event depended on the Mec1 checkpoint kinase as well as on cell cycle arrest triggered by the DNA damage checkpoint network. This phosphorylation event also took place during mitosis of an unperturbed cell cycle. The present finding that S. cerevisiae Scc3 is phosphorylated during mitosis represents a potentially important new regulatory step in cohesin’s mitotic functions.
文摘Severe growth abnormalities including shoot stunting, leaf blade reduction and flower bud failure of Brussels sprout were observed in Poland. The presence of phytoplasma in diseased as well as in healthy looking plants, was demonstrated by nested polymerase chain reaction assay employing phytoplasma universal rRNA primer pairs-P1/P7 followed by R16F2n/R16R2. Products of PCR primed by R 16F2n/R 16R2 primer pair from naturally infected Brussels sprouts were sequenced. Comparison of the obtained 16S rDNAs revealed high nucleotide sequence identity between analyzed phytoplasma isolates (99.8%-100%). They were also nearly identical with the sequences of other phytoplasmas isolates from sub-group 16SrI-B, and they were classified as members of "Candidatus Phytoplasma asteris".
文摘目的探讨人参皂苷20(S)-Rg3抑制卵巢癌细胞增殖和迁移的机制。方法在西安交通大学第一附属医院收集卵巢癌组织19例,正常卵巢组织18例,采用real-time PCR检测lncRNA KRT18P55的表达。将卵巢癌细胞SKOV3和3AO分别分为2组:阴性对照组和20(S)-Rg3组,通过real-time PCR检测KRT18P55的表达水平,CCK-8和平板克隆实验检测SKOV3和3AO细胞增殖能力,Transwell小室法检测SKOV3和3AO细胞迁移能力。将KRT18P55 siRNA分别转染SKOV3和3AO细胞,通过real-time PCR检测KRT18P55的表达水平,CCK-8和平板克隆实验检测SKOV3和3AO细胞增殖能力,Transwell小室法检测SKOV3和3AO细胞迁移能力。癌症基因组学门户网站(cBioPortal for Cancer Genomics,cBioPortal)提取KRT18P55共表达基因数据;基因功能分析数据库(Gene Annotation and Analysis Resource,Metascape)对KRT18P55及其显著相关基因进行功能富集分析;基因表达谱交互式分析(Gene Expression Profiling Interactive Analysis,GEPIA)、阿拉巴马大学癌症数据分析门户(University of Alabama at Birmingham Cancer Data Analysis Portal,UALCAN)在线数据库对筛选的KRT18P55显著相关基因进行表达分析。结果人卵巢癌组织中的KRT18P55表达水平显著高于正常卵巢组织,经20(S)-Rg3处理的卵巢癌细胞SKOV3和3AO的增殖和迁移能力均下降,KRT18P55的表达水平显著降低,差异均具有统计学意义(P<0.05)。敲低KRT18P55后,卵巢癌细胞的增殖和迁移能力下降(P<0.05)。基因本体(Gene Ontology,GO)功能富集分析显示,KRT18P55可能参与中间丝细胞骨架组成、外源性凋亡信号通路等。GEPIA及UALCAN数据库的表达分析显示,KRT18P55相关性最强的两个基因KRT18、KRT8的mRNA及蛋白水平在卵巢癌中均较正常对照显著增高(P<0.05)。结论人参皂苷20(S)-Rg3可能通过降低KRT18P55的表达水平抑制卵巢癌细胞的增殖与迁移,进而抑制卵巢癌的进展。