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目的:探讨雷帕霉素对人食管癌EC1细胞哺乳动物雷帕霉素靶蛋白(mTOR)的表达及细胞生长、凋亡的影响。方法:采用低、中、高浓度的雷帕霉素(100、150、200nmol/L)分别作用于EC1细胞24、48、72h,同时设溶剂对照组和空白对照组,用免疫细胞化学及原位杂交法检测EC1细胞中mTOR蛋白与mRNA的表达,MTT法检测EC1细胞的增殖情况,TUNEL法检测各组作用24h后细胞凋亡情况。结果:雷帕霉素作用24、48与72h,5组细胞mTOR蛋白与mRNA的表达差异均有统计学意义(mTOR蛋白:F=29.273、34.328、41.571,P均<0.001;mTORmRNA:F=34.969、53.614、36.943,P均<0.001),细胞生长抑制率差异亦有统计学意义(F=566.732、51.768和186.022,P均<0.001);雷帕霉素低、中、高浓度组EC1细胞mTOR蛋白与mRNA的表达均低于各对照组,且其表达随雷帕霉素浓度的增加及作用时间的延长而减弱(P<0.05)。雷帕霉素低、中、高浓度组细胞生长抑制率均高于各对照组,且随雷帕霉素浓度的增加和作用时间的延长而升高(P<0.05)。作用24h,5组细胞凋亡指数(AI)差异有统计学意义(F=524.563,P<0.001),雷帕霉素低、中、高浓度组AI均高于各对照组,且随雷帕霉素浓度的增加而增高(P<0.05)。结论:雷帕霉素可能通过抑制EC1细胞mTOR蛋白与mRNA的表达从而抑制EC1细胞的增殖,促进细胞凋亡。
Objective: To investigate the effect of rapamycin on the expression of mammalian target of rapamycin (mTOR) and cell growth and apoptosis in human esophageal EC1 cells. METHODS: EC1 cells were treated with low, medium and high levels of rapamycin (100, 150, 200 nmol / L) for 24, 48 and 72 hours, respectively. The control group and the blank control group were also established. Immunocytochemistry, The hybridization method was used to detect the expression of mTOR protein and mRNA in EC1 cells. The proliferation of EC1 cells was detected by MTT assay. The apoptosis of EC1 cells was detected by TUNEL method. Results: The expressions of mTOR protein and mRNA in 5 groups were all significantly different at 24 h, 48 h and 72 h after rapamycin treatment (mTOR protein: F = 29.273, 34.328, 41.571, P <0.001; mTOR mRNA: F = 34.969 , 53.614, 36.943, P <0.001), and the difference of cell growth inhibition rate was also statistically significant (F = 566.732,51.768 and 186.022, P <0.001) The expression of protein and mRNA were lower than that of the control group, and the expression of protein and mRNA was weakened with the increase of rapamycin concentration and prolongation of action time (P <0.05). The inhibitory rates of rapamycin in low, middle and high concentration groups were higher than those in control group, and increased with rapamycin concentration and prolongation of action time (P <0.05). The apoptotic index (AI) of 24h and 5 groups were statistically significant (F = 524.563, P <0.001). AI in low, middle and high concentration rapamycin groups were higher than those in control group Asmycin concentration increased (P <0.05). Conclusion: Rapamycin may inhibit EC1 cell proliferation and apoptosis by inhibiting the expression of mTOR protein and mRNA in EC1 cells.