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目的运用磷脂酰肌醇3-激酶抑制剂(phosphatidylinositol 3-kinases,PI3-K)LY294002[2-(4-吗啉基)-8-苯基-4氢-1-苯并吡喃-4-酮]作用于胃癌细胞系SGC7901,探讨抑制PI3K/Akt信号转导通路对胃癌细胞化疗敏感性的影响。方法采用MTT比色法,流式细胞术检测5-FU、DDP及ADM单独或联合PI3K抑制剂LY294002对人胃癌细胞SGC7901的抑制率、凋亡率。并分析单独及联合应用LY294002对SGC7901细胞周期的影响。Western-blot检测单独及联合化疗药后P-Akt蛋白在SGC7901细胞中的表达水平。结果单独使用化疗药5-FU、DDP及ADM均可抑制SGC7901细胞增殖、诱导其凋亡。当化疗药与抑制剂联合应用,对细胞的抑制作用明显增强,促凋亡作用增强,与对照组比较(P<0.05)。细胞周期同步分析显示,单独用药均可将SGC7901细胞阻滞于G0/G1期。联合使用抑制剂使处于G0/G1期细胞增加。Western blot显示化疗药上调P-Akt蛋白的表达,联合使用抑制剂后SGC7901细胞P-Akt蛋白的表达与未使用抑制剂比较减弱,差异有统计学意义(P<0.05)。结论阻断PI3K/Akt信号通路可提高化疗药5-FU、DDP及ADM对胃癌细胞株SGC7901的抑制率,凋亡率并使阻滞于G0/G1期细胞增多;LY294002通过阻断PI3K/Akt信号通路,抑制P-Akt蛋白表达,增强化疗药的敏感性;LY294002阻断PI3K/Akt信号通路对5-FU、DDP、ADM治疗胃癌有一定的协同或增强作用。
OBJECTIVE: To investigate the effects of phosphatidylinositol 3-kinases (PI3-K) LY294002 [2- (4-morpholinyl) -8- Ketone] on the gastric cancer cell line SGC7901 to investigate the effect of inhibiting the PI3K / Akt signal transduction pathway on the chemosensitivity of gastric cancer cells. Methods MTT assay and flow cytometry were used to detect the inhibitory rate and apoptosis rate of 5-FU, DDP and ADM alone or in combination with PI3K inhibitor LY294002 on human gastric cancer SGC7901 cells. The effects of LY294002 alone and in combination on SGC7901 cell cycle were analyzed. Western-blot was used to detect the expression of P-Akt protein in SGC7901 cells alone and in combination with chemotherapy. Results The chemotherapeutic agents 5-FU, DDP and ADM alone could inhibit the proliferation and induce the apoptosis of SGC7901 cells. When combined with chemotherapeutic agents and inhibitors, the inhibitory effect on cells was significantly enhanced and the pro-apoptotic effect was enhanced compared with the control group (P <0.05). Cell cycle synchronization analysis showed that SGC7901 cells can be arrested in the G0 / G1 phase alone. Combined use of inhibitors increases G0 / G1 phase cells. Western blot showed that the chemotherapeutic drugs up-regulated the expression of P-Akt protein. The expression of P-Akt protein in SGC7901 cells was weaker than that in unused inhibitor, and the difference was statistically significant (P <0.05). Conclusion The blockade of PI3K / Akt signaling pathway can increase the inhibitory rate and apoptosis rate of gastric cancer cell line SGC7901 by chemotherapeutic agents 5-FU, DDP and ADM, and increase the number of cells arrested in G0 / G1 phase. LY294002 can block PI3K / Akt LY294002 block PI3K / Akt signaling pathway for 5-FU, DDP, ADM treatment of gastric cancer have a certain synergistic or enhanced role.