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目的探讨miR-155提高前列腺癌化疗增敏作用的效应和分子机制。方法转染anti-miR-155,抑制前列腺癌DU145和PC-3细胞中miR-155水平,联合应用顺铂治疗,通过四甲基偶氮唑盐(microculture tetrazolium,MTT)法观察DU145和PC-3细胞的增殖水平,流式细胞仪检测细胞周期和细胞凋亡的变化,蛋白免疫印迹(western blotting)观察Cdc2和cyclin B1蛋白的表达,同时检测caspase-3和caspase-9活性的变化。取对数生长期细胞分对照组、顺铂处理组,anti-miR-155处理组,顺铂联合anti-miR-155处理组,每组n=6。结果与对照组相比,顺铂处理组,anti-miR-155处理组中DU145和PC-3细胞细胞增殖水平显著降低;引起G2/M期细胞阻滞,Cdc2和cyclin B1蛋白表达水平均降低;细胞凋亡率增加,caspase-3和caspase-9活性均提高,组间比较差异均具有统计学意义(P<0.05)。顺铂联合anti-miR-155处理组的细胞增殖水平显著高于对照组;G2/M期细胞阻滞明显,Cdc2和cyclin B1蛋白表达的水平均显著低于对照组;而细胞凋亡率,caspase-3和caspase-9活性等均显著高于对照组,组间比较差异均具有统计学意义(P<0.01)。结论抑制miR-155可增加前列腺癌细胞对顺铂的敏感性,提高顺铂的临床治疗效果,其机制可能与细胞周期和细胞凋亡的进程密切相关。
Objective To investigate the effect and molecular mechanism of miR-155 in enhancing chemosensitivity of prostate cancer. Methods The anti-miR-155 was transfected into DU145 and PC-3 cells to inhibit the expression of miR-155. The combination of cisplatin and MTT was used to detect the expression of DU145 and PC- 3 cells were detected by flow cytometry. The changes of cell cycle and apoptosis were detected by flow cytometry. The expressions of Cdc2 and cyclin B1 protein were detected by Western blotting. The changes of caspase-3 and caspase-9 activity were detected. The cells in logarithmic growth phase were divided into control group, cisplatin treatment group, anti-miR-155 treatment group and cisplatin combined anti-miR-155 treatment group, n = 6 in each group. Results Compared with the control group, the proliferation of DU145 and PC-3 cells in cisplatin-treated group and anti-miR-155 group was significantly decreased, and the cell cycle arrest in G2 / M phase and the protein expression of Cdc2 and cyclin B1 were decreased ; The apoptosis rate increased, the activity of caspase-3 and caspase-9 increased, the differences between the two groups were statistically significant (P <0.05). The cell proliferation of cisplatin combined with anti-miR-155 treatment group was significantly higher than that of the control group; G2 / M phase cell arrest, Cdc2 and cyclin B1 protein levels were significantly lower than the control group; while the apoptosis rate, The activity of caspase-3 and caspase-9 were significantly higher than that of the control group. The difference between the two groups was statistically significant (P <0.01). Conclusion Inhibition of miR-155 can increase the sensitivity of prostate cancer cells to cisplatin and improve the clinical efficacy of cisplatin. The possible mechanism may be related to the cell cycle and the process of apoptosis.