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目的:探讨白藜芦醇诱导胃癌原代细胞发生凋亡的可能性,揭示该凋亡发生与Bcl-2和Bax之间的关系。方法:在体外实验中,采用MTT比色法测定白藜芦醇对胃癌原代细胞的生长抑制率;以透射电镜和TUNEL染色法,定性、定量地研究白藜芦醇与胃癌原代细胞凋亡的关系;通过免疫组织化学法和RT-PCR检测凋亡相关基因bcl-2和bax的表达。结果:白藜芦醇对胃癌原代细胞生长有明显抑制作用,随白藜芦醇浓度增加和培养时间的延长而增强;白藜芦醇诱导胃癌原代细胞出现典型的凋亡细胞形态;TUNEL染色法可见,经白藜芦醇处理24h、48h、72h、96h后,胃癌原代细胞凋亡数明显随时间增加。免疫组化发现经白藜芦醇处理24h、48h、72h、96h后,胃癌原代细胞的Bcl-2蛋白阳性率减少,Bax蛋白阳性率增加。经白藜芦醇处理24、48、72、96h后,RT-PCR检测发现胃癌原代细胞bcl-2和bax的mRNA表达阳性,并显示bcl-2mRNA条带密度随时间的延长而递减,baxmRNA条带密度随时间的延长而增强。结论:白藜芦醇可能通过下调bcl-2的表达和上调bax的表达而诱导胃癌原代细胞发生凋亡。
Objective: To investigate the possibility of apoptosis induced by resveratrol in primary gastric cancer cells and to reveal the relationship between the occurrence of apoptosis and the expression of Bcl-2 and Bax. Methods: In vitro experiments, MTT colorimetric determination of resveratrol on primary gastric cancer cell growth inhibition rate; transmission electron microscopy and TUNEL staining, qualitative and quantitative study of resveratrol and gastric cancer primary cell apoptosis The expression of apoptosis-related genes bcl-2 and bax were detected by immunohistochemistry and RT-PCR. Results: Resveratrol significantly inhibited the growth of primary gastric cancer cells and increased with the increase of resveratrol concentration and incubation time. Resveratrol induced the appearance of typical apoptotic cells in primary gastric cancer cells. TUNEL Staining showed that after resveratrol treatment 24h, 48h, 72h, 96h, the number of primary gastric cancer cells significantly increased with time. Immunohistochemistry showed that after resveratrol treatment for 24h, 48h, 72h, 96h, the positive rate of Bcl-2 protein in primary gastric cancer cells decreased and the positive rate of Bax protein increased. After treated with resveratrol for 24,48,72 and 96 h, the mRNA expression of bcl-2 and bax in primary gastric cancer cells was detected by RT-PCR and showed that the density of bcl-2 mRNA bands decreased with time, the expression of bax mRNA Stripe density increases with time. Conclusion: Resveratrol may induce apoptosis of primary gastric cancer cells by down-regulating bcl-2 expression and up-regulating the expression of bax.