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
AIM： To determine whether midkine （M/O and its truncated form （tMK） contribute to gastric tumorigenesis using in vitro and in vivo models. METHODS： Human MK and tMt（ plasmids were constructed and expressed in BG...AIM： To determine whether midkine （M/O and its truncated form （tMK） contribute to gastric tumorigenesis using in vitro and in vivo models. METHODS： Human MK and tMt（ plasmids were constructed and expressed in BGC823 （a gastric adenocarcinoma cell line） to investigate the effect of over-expressed MK or tMK on cell growth and turmorigenesis in nude mice. RESULTS： The growth of MK-transfected or tMK- transfected cells was significantly increased compared with that of the control cells, and tMK-transfected cells grew more rapidly than MK-transfected cells. The number of colony formation of the cells transfected with MK or tMK gene was larger than the control cells. In nude mice injected with MK-transfected or tMK-transfected cells, visible tumor was observed earlier and the tumor tissues were larger in size and weight than in control animals that were injected with cells without the transfection of either genes. CONCLUSION： Over-expressed MK or tMtC can promote human gastric cancer cell growth in vitro and in vivo, and bMK has greater effect than MK. tMK may be a more promising gene therapeutic target compared with MK for treatment of malignant tumors.展开更多
Various biological indices were used to observe the effects of Tween-80 in combination with hyperthermia at different temperature (39-43℃) for different period of time (20-100 minutes) on human stomach cell line BGC-...Various biological indices were used to observe the effects of Tween-80 in combination with hyperthermia at different temperature (39-43℃) for different period of time (20-100 minutes) on human stomach cell line BGC-823. The results showed that Tween-80obviously reduced the activation energy of BGC-823 cells. Synergistic effect was observed if applied with heat at 39℃ with the increase in temperature and time,the inhibitory effect on the cancer cells was gradually intensified. The lethal rate of BGC-823 cells treated by heat at 41℃ in combination with Tween-80 was around 5.2 times as that treated by hyperthermia alone. The synergistic effect of heat at 41℃ for 100 min. in combination with Tween-80 was equivalent to the effect of 43℃ for 100 min.In other words, the critical temperature for BGC-823 cells was hereby reduced about 2℃. The measurement of membrane mobility, SDH activity etc.also showed that at 41℃ the synergistic effect of hyperthermia in combination with Tween-80 was the best, it exceeding the single effect of these factors in combination and showing effect of multiplication. The synergistic effect of heat at 41℃ in combination with Tween-80 was higher than that of heat at 41℃ in combination with MMC. It also demonstrated that the specific and sustained action on the inhibition of cancer cells did exist. These studies suggested that the synergistic mechanism of Tween-80 and hyperthermia probably was that both of them acted on the cell membrane system. Supported by The National Natural Science Fund of China (No. 3860948)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
背景与目的：探讨丹参酮ⅡA（tanshinoneⅡA，TanⅡA）对人胃癌BGC-823细胞增殖及凋亡的影响。材料与方法：采用0～10μg／ml Tan ⅡA分别作用于BGC-823细胞48h及72h，MTT比色法观察药物对细胞生长的抑制效应；2、5、10μg／ml TanⅡA...背景与目的：探讨丹参酮ⅡA（tanshinoneⅡA，TanⅡA）对人胃癌BGC-823细胞增殖及凋亡的影响。材料与方法：采用0～10μg／ml Tan ⅡA分别作用于BGC-823细胞48h及72h，MTT比色法观察药物对细胞生长的抑制效应；2、5、10μg／ml TanⅡA分别作用于细胞72h，应用HE染色、DNA琼脂糖凝胶电泳、流式细胞术分析药物对细胞凋亡的影响。结果：TanⅡA明显抑制BGC-823细胞增殖、诱导细胞凋亡，镜下可见BGC-823细胞明显的凋亡特征性改变；5、10μg／ml Tan ⅡA处理组细胞DNA电泳可见典型的“梯状”条带；FCM结果显示5、10μg／ml TanⅡA处理组细胞，凋亡率分别为（20．60±1．84）％、（31．03±1．47）％，与对照组（5．23±0．39）％比较差异有统计学意义（P〈0．01）。结论：在一定条件下，TanⅡA可抑制人胃癌BGC-823细胞增殖，诱导其凋亡。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 The Scientific Research Fund of Graduate School of Nanjing University, the Fund for Key Program of Ministry of Education, No. 02111the 985-Ⅱ Program of Nanjing University
文摘AIM： To determine whether midkine （M/O and its truncated form （tMK） contribute to gastric tumorigenesis using in vitro and in vivo models. METHODS： Human MK and tMt（ plasmids were constructed and expressed in BGC823 （a gastric adenocarcinoma cell line） to investigate the effect of over-expressed MK or tMK on cell growth and turmorigenesis in nude mice. RESULTS： The growth of MK-transfected or tMK- transfected cells was significantly increased compared with that of the control cells, and tMK-transfected cells grew more rapidly than MK-transfected cells. The number of colony formation of the cells transfected with MK or tMK gene was larger than the control cells. In nude mice injected with MK-transfected or tMK-transfected cells, visible tumor was observed earlier and the tumor tissues were larger in size and weight than in control animals that were injected with cells without the transfection of either genes. CONCLUSION： Over-expressed MK or tMtC can promote human gastric cancer cell growth in vitro and in vivo, and bMK has greater effect than MK. tMK may be a more promising gene therapeutic target compared with MK for treatment of malignant tumors.
文摘Various biological indices were used to observe the effects of Tween-80 in combination with hyperthermia at different temperature (39-43℃) for different period of time (20-100 minutes) on human stomach cell line BGC-823. The results showed that Tween-80obviously reduced the activation energy of BGC-823 cells. Synergistic effect was observed if applied with heat at 39℃ with the increase in temperature and time,the inhibitory effect on the cancer cells was gradually intensified. The lethal rate of BGC-823 cells treated by heat at 41℃ in combination with Tween-80 was around 5.2 times as that treated by hyperthermia alone. The synergistic effect of heat at 41℃ for 100 min. in combination with Tween-80 was equivalent to the effect of 43℃ for 100 min.In other words, the critical temperature for BGC-823 cells was hereby reduced about 2℃. The measurement of membrane mobility, SDH activity etc.also showed that at 41℃ the synergistic effect of hyperthermia in combination with Tween-80 was the best, it exceeding the single effect of these factors in combination and showing effect of multiplication. The synergistic effect of heat at 41℃ in combination with Tween-80 was higher than that of heat at 41℃ in combination with MMC. It also demonstrated that the specific and sustained action on the inhibition of cancer cells did exist. These studies suggested that the synergistic mechanism of Tween-80 and hyperthermia probably was that both of them acted on the cell membrane system. Supported by The National Natural Science Fund of China (No. 3860948)
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。
文摘背景与目的：探讨丹参酮ⅡA（tanshinoneⅡA，TanⅡA）对人胃癌BGC-823细胞增殖及凋亡的影响。材料与方法：采用0～10μg／ml Tan ⅡA分别作用于BGC-823细胞48h及72h，MTT比色法观察药物对细胞生长的抑制效应；2、5、10μg／ml TanⅡA分别作用于细胞72h，应用HE染色、DNA琼脂糖凝胶电泳、流式细胞术分析药物对细胞凋亡的影响。结果：TanⅡA明显抑制BGC-823细胞增殖、诱导细胞凋亡，镜下可见BGC-823细胞明显的凋亡特征性改变；5、10μg／ml Tan ⅡA处理组细胞DNA电泳可见典型的“梯状”条带；FCM结果显示5、10μg／ml TanⅡA处理组细胞，凋亡率分别为（20．60±1．84）％、（31．03±1．47）％，与对照组（5．23±0．39）％比较差异有统计学意义（P〈0．01）。结论：在一定条件下，TanⅡA可抑制人胃癌BGC-823细胞增殖，诱导其凋亡。
