Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segm...Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segment for Bcl-2 gene was designed and synthesized,then was induced into gastric cancer BGC 823 cells by liposome transfection.Bcl-2 protein expression was detected by Western Blotting.After X radiation,flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity. Result:After the transfection of Bcl-2 siRNA,the positive expression rate of Bcl-2 protein in BGC823 cells was（35.45±2.35）%.Compared with the control group and negative siRNA transfection group,the rate was significantly decreased（P【0.01）.The apoptosis rate of BGC823-RNAi cell was（10.81±0.91）%,which was significantly higher than the control group and negative siRNA transfection group（P【0.01）.After 48h X radiation,the apoptosis rate of BGC823-RNAi was（28.91±1.40）%,which was significantly higher than the control group and the group without radiation （P【0.01）.During clone forming assay D<sub>0</sub>,D<sub>4</sub> and SF<sub>2</sub> values in Bcl-2 siRNA1 transfection group were all lower than those in the control group.The radiosensitivity ratio was 1.28（the ratio of D<sub>0</sub>） and 1.60（the ratio of D<sub>4</sub>）.Conclusions:Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene,enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells,having good clinical application perspective.展开更多
[Objectives]This study aimed to investigate the inhibiting effect of Obazema on proliferation of gastric cancer BGC-823 cells and its mechanism.[Methods]BGC-823 cells were treated with high,medium and low concentratio...[Objectives]This study aimed to investigate the inhibiting effect of Obazema on proliferation of gastric cancer BGC-823 cells and its mechanism.[Methods]BGC-823 cells were treated with high,medium and low concentrations of drug-containing serum(0.316%,0.158%and 0.079%)for 0,48,72 and 96 h,respectively.Then,the proliferation of the cells was detected with CCK-8 method,and the expression of related proteins,B lymphocytoma-2(Bcl-2),phosphorylated protein kinase B(p-Akt),protein kinase B(Akt)and glyceraldehyde-3-phosphate dehydrogenase(GAPDH),was detected using Western blotting.[Results]The proliferation of the BGC-823 cells was significantly inhibited with different doses of Boenninghausenia albiflora(Hook.)Reichb.ex Meisn.var.albiflora(CH)and B.sessilicarpa Lévl.(S)(P<0.01),in dose-dependent and time-dependent manners.The inhibition of high-dose S on cell proliferation was similar to that of CTX 48 h after administration;the inhibition of high-dose CH on cell proliferation was significantly stronger than that of CTX(P<0.01);different doses of drug administration groups significantly inhibited the expression of p-Akt and Bcl-2 in the BGC-823 cells;the inhibition of high-dose CH on the expression of P-Akt and Bcl-2 and the inhibition of medium-dose CH on the expression of Bcl-2 were significantly stronger than that of CTX(P<0.05,P<0.01),in a certain dose-dependent manner;at the same dose,the inhibition of CH on the expression of the proteins was stronger than that of S(P<0.05,P<0.01);administration of S and CH significantly inhibited the expression of GAPDH compared with CTX(P<0.05,P<0.01).[Conclusions]Obazema has the capacity to inhibit the proliferation of BGC-823 cells.The mechanism may be achieved by inhibiting the expression of p-Akt and Bcl-2,and GAPDH may be the target gene of its anti-tumor mechanism.The inhibiting effect of CH on BGC-823 cells was more significantly than that of S.展开更多
The effect of LaCit on cell proliferation, DNA synthesize, cell cycle, cell membrane fluidity, growth in soft agar in human lung cancer cells PG and human gastric carcinoma cells BGC 823 was studied by MTT(3 (4,5 d...The effect of LaCit on cell proliferation, DNA synthesize, cell cycle, cell membrane fluidity, growth in soft agar in human lung cancer cells PG and human gastric carcinoma cells BGC 823 was studied by MTT(3 (4,5 dimethylthiazol 2 yl) 2,5 diphenytetrazolium bromide) assay, 3 H TdR incorporation, flow cytometry, fluorescence polarization assay, and soft agar culture. The results indicate that when the concentrations of LaCit are 0.05, 0.1, 0.5 and 1 mmol·L -1 , it inhibits PG cells proliferation. When the concentrations are 0.5 and 1 mmol·L -1 LaCit inhibits PG cells 3 H TdR incorporation, and decreases the proportion of PG cells in S phase while increases the proportion of PG cells in G 1 phase and decreases the membrane fluidity of PG cells. But LaCit has no effect on BGC 823 cells except for cell membrane fluidity and no effect on 2BS cells.展开更多
目的研究香加皮水提取物(CPE)诱导人胃癌细胞BGC-823凋亡及其作用机制。方法采用G iem sa染色观察细胞凋亡形态学变化;电子显微镜观察凋亡细胞的超微结构变化;流式细胞术和琼脂糖凝胶电泳方法检测BGC-823细胞凋亡率、细胞周期和细胞凋亡...目的研究香加皮水提取物(CPE)诱导人胃癌细胞BGC-823凋亡及其作用机制。方法采用G iem sa染色观察细胞凋亡形态学变化;电子显微镜观察凋亡细胞的超微结构变化;流式细胞术和琼脂糖凝胶电泳方法检测BGC-823细胞凋亡率、细胞周期和细胞凋亡的DNA水平变化;RT-PCR方法检测细胞凋亡相关基因bcl-2、bax和surv iv in mRNA表达水平变化;免疫细胞化学方法检测bcl-2、bax和surv iv in蛋白表达的变化。结果经CPE作用后,人胃癌细胞BGC-823出现明显的细胞凋亡形态学变化及超微结构改变,细胞DNA琼脂糖凝胶电泳呈现梯形图。经250μg/mL CPE处理48 h后,多数BGC-823细胞被阻滞在G2/M期,而且细胞发生明显的凋亡变化,BGC-823细胞凋亡率可达18.9%。CPE可抑制BGC-823细胞bcl和surv iv in mRNA及蛋白的表达,促进baxmRNA及蛋白的表达。CPE可明显延长S180荷瘤小鼠生存期,且具有剂量依赖性。结论CPE通过阻滞BGC-823细胞于G2/M期及诱导BGC-823细胞凋亡发挥抗肿瘤作用,其作用机制与抑制细胞的bcl-2和surv iv in基因mRNA及蛋白表达、促进bax基因和蛋白的表达有关。展开更多
文摘Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segment for Bcl-2 gene was designed and synthesized,then was induced into gastric cancer BGC 823 cells by liposome transfection.Bcl-2 protein expression was detected by Western Blotting.After X radiation,flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity. Result:After the transfection of Bcl-2 siRNA,the positive expression rate of Bcl-2 protein in BGC823 cells was（35.45±2.35）%.Compared with the control group and negative siRNA transfection group,the rate was significantly decreased（P【0.01）.The apoptosis rate of BGC823-RNAi cell was（10.81±0.91）%,which was significantly higher than the control group and negative siRNA transfection group（P【0.01）.After 48h X radiation,the apoptosis rate of BGC823-RNAi was（28.91±1.40）%,which was significantly higher than the control group and the group without radiation （P【0.01）.During clone forming assay D<sub>0</sub>,D<sub>4</sub> and SF<sub>2</sub> values in Bcl-2 siRNA1 transfection group were all lower than those in the control group.The radiosensitivity ratio was 1.28（the ratio of D<sub>0</sub>） and 1.60（the ratio of D<sub>4</sub>）.Conclusions:Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene,enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells,having good clinical application perspective.
基金Supported by National Key Technology Research and Development Program(2018FYC1708000)Basic Research Project for Application of Science and Technology in Sichuan Province(2017JY0274)Science and Technology Research Project of Sichuan Traditional Chinese Medicine Administration(2018JC028).
文摘[Objectives]This study aimed to investigate the inhibiting effect of Obazema on proliferation of gastric cancer BGC-823 cells and its mechanism.[Methods]BGC-823 cells were treated with high,medium and low concentrations of drug-containing serum(0.316%,0.158%and 0.079%)for 0,48,72 and 96 h,respectively.Then,the proliferation of the cells was detected with CCK-8 method,and the expression of related proteins,B lymphocytoma-2(Bcl-2),phosphorylated protein kinase B(p-Akt),protein kinase B(Akt)and glyceraldehyde-3-phosphate dehydrogenase(GAPDH),was detected using Western blotting.[Results]The proliferation of the BGC-823 cells was significantly inhibited with different doses of Boenninghausenia albiflora(Hook.)Reichb.ex Meisn.var.albiflora(CH)and B.sessilicarpa Lévl.(S)(P<0.01),in dose-dependent and time-dependent manners.The inhibition of high-dose S on cell proliferation was similar to that of CTX 48 h after administration;the inhibition of high-dose CH on cell proliferation was significantly stronger than that of CTX(P<0.01);different doses of drug administration groups significantly inhibited the expression of p-Akt and Bcl-2 in the BGC-823 cells;the inhibition of high-dose CH on the expression of P-Akt and Bcl-2 and the inhibition of medium-dose CH on the expression of Bcl-2 were significantly stronger than that of CTX(P<0.05,P<0.01),in a certain dose-dependent manner;at the same dose,the inhibition of CH on the expression of the proteins was stronger than that of S(P<0.05,P<0.01);administration of S and CH significantly inhibited the expression of GAPDH compared with CTX(P<0.05,P<0.01).[Conclusions]Obazema has the capacity to inhibit the proliferation of BGC-823 cells.The mechanism may be achieved by inhibiting the expression of p-Akt and Bcl-2,and GAPDH may be the target gene of its anti-tumor mechanism.The inhibiting effect of CH on BGC-823 cells was more significantly than that of S.
文摘The effect of LaCit on cell proliferation, DNA synthesize, cell cycle, cell membrane fluidity, growth in soft agar in human lung cancer cells PG and human gastric carcinoma cells BGC 823 was studied by MTT(3 (4,5 dimethylthiazol 2 yl) 2,5 diphenytetrazolium bromide) assay, 3 H TdR incorporation, flow cytometry, fluorescence polarization assay, and soft agar culture. The results indicate that when the concentrations of LaCit are 0.05, 0.1, 0.5 and 1 mmol·L -1 , it inhibits PG cells proliferation. When the concentrations are 0.5 and 1 mmol·L -1 LaCit inhibits PG cells 3 H TdR incorporation, and decreases the proportion of PG cells in S phase while increases the proportion of PG cells in G 1 phase and decreases the membrane fluidity of PG cells. But LaCit has no effect on BGC 823 cells except for cell membrane fluidity and no effect on 2BS cells.
文摘目的研究香加皮水提取物(CPE)诱导人胃癌细胞BGC-823凋亡及其作用机制。方法采用G iem sa染色观察细胞凋亡形态学变化;电子显微镜观察凋亡细胞的超微结构变化;流式细胞术和琼脂糖凝胶电泳方法检测BGC-823细胞凋亡率、细胞周期和细胞凋亡的DNA水平变化;RT-PCR方法检测细胞凋亡相关基因bcl-2、bax和surv iv in mRNA表达水平变化;免疫细胞化学方法检测bcl-2、bax和surv iv in蛋白表达的变化。结果经CPE作用后,人胃癌细胞BGC-823出现明显的细胞凋亡形态学变化及超微结构改变,细胞DNA琼脂糖凝胶电泳呈现梯形图。经250μg/mL CPE处理48 h后,多数BGC-823细胞被阻滞在G2/M期,而且细胞发生明显的凋亡变化,BGC-823细胞凋亡率可达18.9%。CPE可抑制BGC-823细胞bcl和surv iv in mRNA及蛋白的表达,促进baxmRNA及蛋白的表达。CPE可明显延长S180荷瘤小鼠生存期,且具有剂量依赖性。结论CPE通过阻滞BGC-823细胞于G2/M期及诱导BGC-823细胞凋亡发挥抗肿瘤作用,其作用机制与抑制细胞的bcl-2和surv iv in基因mRNA及蛋白表达、促进bax基因和蛋白的表达有关。
