黏液瘤病毒对卵巢癌细胞增殖的抑制作用及其机制探讨  

Effect of myxomavirus on the proliferation of ovarian cancer cells and its mechanism

作　　者：赖蔚菁[1] 贺锦曦[1] 尤嘉萍[1] 黄晔[2] 吴美秀 

机构地区：[1]福建医科大学附属泉州第一医院妇产科,福建泉州362000 [2]福建医科大学省立临床医学院妇产科,福建福州350001 [3]温州市中心医院妇产科,浙江温州325000

出　　处：《肿瘤》2016年第11期1196-1202,共7页Tumor

摘　　要：目的:探讨黏液瘤病毒(myxomavirus,MV)感染对人卵巢癌SKOV3细胞增殖的影响及其分子机制。方法:体外培养人卵巢癌SKOV3细胞,给予MV感染处理;同时,以灭活病毒处理作为阴性对照组,以RPMI 1640培养液处理作为空白对照组。采用CCK-8法检测各组卵巢癌SKOV3细胞的增殖情况,实时荧光定量PCR法检测各组细胞中Bcl-2和survivin的m RNA水平,FCM法分析各组细胞周期分布,蛋白质印迹法检测各组细胞中细胞外调节蛋白激酶1/(2extracellular regulated protein kinase 1/2,ERK1/2)、蛋白激酶B(protein kinase B,PKB,又称Akt)、磷酸化ERK1/2、磷酸化Akt、Bcl-2和survivin的蛋白表达水平,并采用比色法测定caspase-3和caspase-8的活性。结果:与阴性对照组和空白对照组相比,MV能够明显抑制人卵巢癌SKOV3细胞的增殖和细胞周期进程(P值均<0.05)。MV作用SKOV3细胞96 h后,抗凋亡蛋白Bcl-2和survivin的m RNA及蛋白表达水平均明显下调(P值均<0.05),增殖相关信号通路中ERK1/2和Akt的磷酸化水平均明显降低(P值均<0.05),而caspase-3和caspase-8的活性均明显升高(P值均<0.05)。结论 :MV能明显抑制卵巢癌细胞的增殖,其作用机制可能与阻滞细胞周期,下调抗凋亡蛋白Bcl-2和survivin的表达,提高caspase-3和caspase-8的活性,并抑制增殖相关信号通路中ERK1/2和Akt的磷酸化有关。Objective： To investigate the effect of myxomavirus （MV） on the proliferation of human ovarian cancer SKOV3 cells, and its molecular mechanism. Methods： The human ovarian cancer SKOV3 cells were cultured in vitro and infected with MV. At the same time, SKOV3 cells infected with inactivated virus or only cultured with RPMI 1640 medium were used as the negative control group or the blank group, respectively. The proliferation of SKOV3 cells in the three groups was determined by CCK-8 assay. The mRNA levels of Bcl-2 and survivin were detected by real-time fluorescent quantitative PCR. The cell cycle distribution was analyzed by FCM. The expressions of total extracellular regulated protein kinase 1/2 （ERK1/2）, phosphorylated ERK1/2 （p-ERK1/2）, Akt, p-Akt, Bcl-2 and survivin proteins were measured by Western blotting. The activities of caspase-3 and caspase-8 were also quantified by colorimetric method. Results： Compared with the negative control group and the blank group, MV significantly inhibited the proliferation and cell cycle progression of human ovarian cancer SKOV3 cells （all P 〈 0.05）. After SKOV3 cells were infected with MV for 96 h, the mRNA and protein expressions of Bcl-2 and survivin were significantly down-regulated （both P 〈 0.05）, while the phosphorylation levels of ERK1/2 and Akt were significantly decreased （both P 〈 0.05）, but the activities of caspase-3 and caspase-8 were obviously enhanced （both P 〈 0.05）. Conclusion： MV can inhibit the proliferation of ovarian cancer cells, and its mechanism may be related to blocking cell cycle progression, down-regulating the expressions of anti-apoptotic proteins Bcl-2 and survivin, increasing the activation of caspase-3 and caspase-8, and inhibiting the phosphorylation of ERK and Akt in proliferation-related signal pathway.

关 键 词：卵巢肿瘤 痘苗病毒 细胞增殖 凋亡调节蛋白质类 黏液瘤病毒 

分 类 号：R737.31[医药卫生—肿瘤]