摘要
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 explore the effects of miR-155 on chemotherapy sensitivity enhancement of prostate cancer in vitro.Methods The anti-miR-155 was transfected to inhibit levels of miR-155 in prostate cancer DU145 and PC-3cells after anti-miR-155 transfection combined with cisplatin treatment.Levels of DU145 and PC-3 cell proliferation were observed by microculture tetrazolium（MTT）,meanwhile flow cytometry were performed to evaluate the cell cycle and apoptosis.Western boltting analysis was used to detect Cdc2 and cyclin B1 protein expression,and the variation of caspase-3and caspase-9 activity.Cells in logarithmic growth phase were divided into control group,cisplatin group,anti-miR-155 group and cisplatin＋anti-miR-155 group（n=6）.Results Compared with control group,cell proliferation levels of DU145 and PC-3 in both of anti-miR-155 group and cisplatin group were obviously inhibited.Cdc2 and cyclin B1 protein expression was markedly decreased with cell blocking at G2/M phase.In addition,apoptosis rate,caspase-3 and caspase-9 activities were enhanced in anti-miR-155 group and cisplatin group which were significantly different（P〈0.05）.The cell proliferation levels in cisplatin＋anti-miR-155 group were prominently higher than those in control group,however,Cdc2 and cyclin B1 protein expression levels were obviously lower than those in control group with cell blocking at G2/M phase.The apoptosis rate,caspase-3 and caspase-9 activities were higher than those in control group,which were significantly different（P〈0.05）.Conclusion The inhibition of miR-155 can enhance the chemosensitivity of cisplatin in prostate cancer and increse clinical effects under cisplatin treatment.Its mechanism may be closely related to the process of cell cycle and apoptosis.
出处
《华南国防医学杂志》
CAS
2016年第7期421-425,共5页
Military Medical Journal of South China
基金
国家自然科学基金项目(81301303)
