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目的探讨5-氮脱氧胞苷(5-aza-dC)与雷帕霉素(RPM)对人结直肠癌细胞的联合作用及机制。方法应用 MTY 比色法检测5-aza-dC 与 RPM 对3种人结直肠癌细胞株的生长抑制作用;应用流式细胞术检测两者对细胞周期的调控;应用免疫印迹法检测两者对于 mTOR 信号转导通路相关蛋白的影响。结果 5-aza-dC 与 RPM 对人结直肠癌细胞株 SW480、HT29和 HCT116均有生长抑制作用,在相同作用条件下,5-aza-dC 与 RPM 的联合抑制效果明显优于两者单独用药。5-aza-dC 与 RPM单独或联合用药对3种细胞的生长周期均具有调控作用(P<0.05),但作用位点不尽相同。5-aza-dC与 RPM 可下调 mTOR、p70s6K、4E-BP1蛋白的磷酸化水平,对蛋白的总体水平无明显影响,以两者的联合作用最为显著。结论 5-aza-dC 与 RPM 可联合抑制人结直肠癌细胞株 SW480、HT29和 HCT116的生长,并阻滞其细胞周期进程。其作用机制可能与影响 mTOR 信号转导通路蛋白磷酸化水平有关。
Objective To investigate the combined effects of 5-aza-dC and rapamycin on human colorectal cancer cells and its mechanism. Methods MTT assay was used to detect the inhibitory effect of 5-aza-dC and RPM on the growth of three human colorectal cancer cell lines. Flow cytometry was used to detect the cell cycle regulation. Western blotting was used to detect the effect of both mTOR signaling pathway related proteins. Results Both 5-aza-dC and RPM could inhibit the growth of human colorectal cancer cell lines SW480, HT29 and HCT116. Under the same conditions, the combination of 5-aza-dC and RPM had better inhibitory effect than the two alone . 5-aza-dC alone or in combination with RPM could regulate the growth cycle of all three kinds of cells (P <0.05), but the sites of action were different. 5-aza-dC and RPM could down-regulate the phosphorylation levels of mTOR, p70s6K and 4E-BP1 protein, but had no significant effect on the overall protein level. Conclusions 5-aza-dC and RPM can inhibit the growth of human colorectal cancer cell lines SW480, HT29 and HCT116 and block the cell cycle progression. Its mechanism may be related to the impact of mTOR signal transduction pathway protein phosphorylation level.