摘要
目的：研究 VEGF - ERK 信号通路在软坚散结胶囊含药血清诱导胃癌 SGC7901细胞凋亡过程中的变化，探讨软坚散结胶囊含药血清调控胃癌细胞凋亡的机制。方法制备软坚散结胶囊含药血清；激光共聚焦显微镜观察软坚散结胶囊含药血清处理后胃癌 SGC7901细胞形态变化；流式细胞仪检测软坚散结胶囊含药血清处理后胃癌 SGC7901细胞凋亡率；Western Blot 检测软坚散结胶囊含药血清对人胃癌 SGC7901细胞 ERK、P - ERK 及 VEGF 蛋白表达影响。结果软坚散结胶囊含药血清处理细胞后，激光共聚焦显微镜下见：阴性对照组以正常活性细胞为主，Annexin V 和 PI 均为低染，细胞膜完整，细胞形态正常；低剂量组可见 Annexin V 高染、PI 低染，细胞膜呈绿色荧光，细胞核着色浅或不着色；中剂量组可见 Annexin V 高染、PI 低染，细胞膜呈绿色荧光，细胞核呈桔红色荧光；高剂量组 PI 高染，细胞核呈红色荧光，出现部分细胞的细胞膜破裂、细胞核呈大小不等形态不规则的碎片状，为典型的细胞凋亡形态特征。流式细胞仪检测经阴性对照组及低中高剂量组含药血清处理的 SGC7901细胞后，各组细胞凋亡率分别为：（6.053±0.223）％、（9.117±1.583）％、（23.663±3.428）％、（37.707±1.328）％，细胞凋亡率随软坚散结胶囊含药血清剂量的增加而升高。Western Blot 检测发现 SGC7901细胞中 VEGF 蛋白水平随含药血清剂量的增加而降低（ P ﹤0.05）；ERK 蛋白表达无明显变化（ P ﹥0.05），P - ERK（ERK 磷酸化）蛋白水平随含药血清剂量增加而降低（ P ﹤0.05）。结论软坚散结胶囊含药血清能诱导人胃癌 SGC7901细胞凋亡；其作用机制可能是通过抑制 VEGF 蛋白表达，调控 ERK 信号通路，抑制 ERK 磷酸化（p - ERK）表达进而促进胃癌细胞凋亡。
Abstract
Objective To study the changes of VEGF - ERK signal path in the apoptosis of SGC7901 cells of gastric cancer induced by serum drug of ruanjiansanjie capsules(drug serum)and explore the mechanism on the regulation of cell apoptosis in gastric cancer. Methods The drug serum was prepared. Confocal microscopy was used to detect the morphological changes of SGC7091 cell. FCM was used to detect SGC7091 apoptosis after treatment. Western Blot was used to detect ERK,P - ERK and VEGF protein ex-pressions of SGC7901 cells. Results Confocal microscopy showed that the normal active cells were predomi-nated in the negative control group,Annexinv and PI staining was lower,cytomembrane was complete and the morphology was normal. In the low - dose group,Annexin V was highly stained and PI staining was lower,cy-tomembrane was of green fluorescence and the nucli were colored light or not colored. In the middle - dose group,Annexin V was highly stained and PI staining was lower,cytomembrane was of green fluorescence and the nucli were of nacarat one. In the high - dose group,PI was highly stained,the nucli were colored red,the membrane was ruptured and the size of nucli was fragmented,presenting the typical apoptotic characters. FCM showed that after treatment,the apoptotic rates were 6. 053 ± 0. 223% ,9. 117 ± 1. 583% ,23. 663 ± 3. 428%and 37. 707 ± 1. 328% in each group. It was increased with the dose increase of drug serum. Western Blot found that P - ERK expression was significantly decreased(P ﹤0.05),VEGF expression in SGC7901 was reduced <br> with the dose of drug serum increased(P ﹤ 0. 05). The change of ERK protein expression was not significant (P ﹥ 0. 05). The protein level of P - ERK was reduced while the dose of drug serum increased(P ﹤ 0. 05).Conclusion The serum drug induces SGC7901apoptosis of human gastric cancer cell,which may be through the inhibition of VEGF protein expression,the regulation of ERK signal path and the suppression of P - ERK expression.
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维普
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