MicroRNA-210对卵巢癌细胞生长及放射敏感性的影响

摘要

目的:探讨 MicroRNA-210(miR-210)在卵巢癌细胞生长过程中的作用及其与放射治疗敏感性的关系,阐明miR-210对卵巢癌发展和治疗的影响及可能的分子机制.方法:体外培养人卵巢癌细胞株 OVCAR3和SKOV3,应用细胞转染法分别将miR-210 mimic和抗miR-210抑制剂转染至 OVCAR3和 SKOV3细胞中,并利用 Real-time PCR法进行鉴定,得到miR-210过表达和低表达卵巢癌细胞模型.将2种细胞分别分为对照组、miR-210过表达组和miR-210低表达组,采用MTT法检测各组细胞增殖活性;采用不同剂量(30、50和100 Gy)电离辐射照射对照组和miR-210过表达组细胞,采用 MTT法检测各组细胞增殖活性;采用 Western blotting法检测50 Gy照射剂量组卵巢癌细胞中凋亡相关蛋白表达水平.所有实验细胞培养采用3复孔,并重复3次.结果:与对照组比较,miR-210过表达组细胞增殖活性升高,miR-210低表达组细胞增殖活性降低(P<0.05);在给予电离辐射照射后,与对照组比较,miR-210过表达组细胞增殖活性升高(P<0.05);且凋亡相关蛋白 BAX在2株细胞中表达水平均明显下降,而BCL-2表达水平均明显升高.结论:miR-210可促进卵巢癌细胞的生长,同时通过抑制凋亡作用降低卵巢癌细胞对放射治疗的敏感性.%Objective:To investigate the role of MicroRNA-210(miR-210)in the growth process of ovarian cancer cells and its relationship with radiosensitivity,and to elucidate the effect of miR-210 on the development and treatment of ovarian cancer the possible molecular mechanism.Methods:The human ovarian cancer OVCAR3 and SKOV3 cells were cultured in vitro,miR-210 mimic and anti miR-210 inhibitors were transfected into the human ovarian cancer cell lines OVCAR3 and SKOV3,respectively,by cell transfection,and identified by Real-time PCR method;miR-210 overexpression and low expression of ovarian cancer cell models were obtained.The two kinds of cells were divided into control group,miR-210 overexpression model group and miR-210 low expression model group,and the cell proliferation activity was detected by MTT.The cells in control group and miR-210 overexpression model group were irradiated with different doses(30,50,and 100 Gy)of ionizing radiation,and the cell proliferation activity in each group was detected by MTT method.Western blotting method was used to detect the expression levels of apoptosis-related proteins in the ovarian cancer cells exposed to 50 Gy radiation dose.All the experimental cells were cultured with three double holes and repeated three times.Results:Compared with control group,the proliferation activity of the cells in miR-210 overexpression ovarian cancer cells group was enhanced,and the proliferation activity of the cells in miR-210 low expression cells group was reduced(P<0.05).After ionizing radiation,the proliferation activity of ovarian cancer cells in miRNA-210 overexpression group was enhanced compared with control group(P<0.05);the expression levels of apoptosis-related protein BAX in OVCAR3 and SKOV3 cells were significantly decreased,while the expression level of BCL-2 were significantly increased. Conclusion:miR-210 can promote the growth of ovarian cancer cells,and reduce the sensitivity of ovarian cancer cells to radiation therapy by inhibiting the apoptosis.