吗啡对胃癌细胞系HGC27细胞增殖的影响及其机制

目的探讨吗啡对人胃癌细胞系HGC27细胞增殖的影响及其机制。方法体外培养的HGC27细胞,用不同浓度吗啡处理24、48、72 h,四甲基偶氮唑蓝（MTT）法测定其对HGC27细胞增殖的影响;倒置显微镜下观察细胞的形态学变化;实验组用吗啡（浓度分别为0.05、0.1、0.2μmol/L）处理HGC27细胞48 h,对照组加入不含药物的培养基,分别采用流式细胞仪检测细胞凋亡、分光光度计检测细胞凋亡蛋白酶-3（Casepase-3）相对活性、western blot法检p16、bcl-2、bax基因表达情况。结果吗啡（浓度0.025～0.4μmol/L）能抑制HGC27细胞的增殖,呈剂量和时间依赖性;倒置显微镜下可观察到典型的细胞凋亡形态;吗啡（浓度分别为0.05、0.1、0.2μmol/L）作用HGC27细胞48 h后,细胞凋亡率分别为9.4%、11.5%、21.4%,而对照组凋亡率仅为2.2%;Casepase-3相对活性显著增加,处理后Casepase-3相对活性分别为（1.32±0.08）、（1.85±0.06）和（2.45±0.07）,对照组为（0.92±0.07）;p16和bax蛋白表达量升高,而bcl-2蛋白表达量降低,呈剂量依赖性。结论吗啡能抑制HGC27细胞的增殖并诱导其凋亡,其作用机制可能与上调p16表达,进而引起bax蛋白表达量升高,而bcl-2蛋白表达量降低,并增加细胞Casepase-3活性有关。

Effects of morphine on cell proliferation in human gastric cancer HGC27 cells and its mechanism

Objective To investigate the effect of morphine on cell proliferation in gastric cancer line HGC27 cells and discuss its possible mechanism. Methods HGC27 cells were cultured in vitro. After treatment by morphine at different concentrations respectively at different time, the cell survival was determined by the MTT method. The changes of cell morphology were observed by inverted microscope. Apoptosis was detected by flow cytometry. The relative activity of caspase-3 was monitored by spectrophotometer. The changes of protein expression of p16, bcl-2 and bax were detected by western blot. Results From the data of MTT, the cell proliferation of human gastricl cancer HGC27 cells was inhibited by morphine in a dose-dependent and time-dependent manner. Typical apoptosis morphology of HGC27 cells was observed by inverted microscope. Flow cytometry assays showed that morphine significantly induced apoptosis in HGC27 cells. After treated with morphine, the apoptosis rate of HGC27 cells was 9.4%, 11.5% and 21.4% respectively,which showed an obvious concentration-effect relationship. The relative activity of caspase-3 of morphine group was（1.32±0.08）,（1.85±0.06） and（2.45±0.07） respectively, which of control group was（0.92±0.07）. The data of western blot showed that morphine up-regulated p16 and bax and down- regulated bcl-2 expression in a dose-dependent manner.Conclusion Morphine can inhibit the proliferation of HGC27 cells and induce apoptosis, and the mechanism of morphine on apoptosis may be related to the up-regulation of p16 expression, thus results up-regulation of bax and downregulation of bcl-2, as well as the increase of relative activity of caspase-3.