NK4基因转染对人卵巢癌细胞SKOV-3生物学特性的影响  

Effects of NK4 on biological behaviors of human ovarian cancer cell line SKOV-3

作　　者：王冬冬[1] 霍乃晨[1] 耿东华[2] 杨志亮[3] 蔡峰[4] 尹少尉[1] 张淑兰[1] 

机构地区：[1]中国医科大学附属盛京医院妇产科,辽宁沈阳110004 [2]中国医科大学附属盛京医院普外科,辽宁沈阳110004 [3]中国医科大学附属第一医院儿科,辽宁沈阳110001 [4]中国医科大学附属第一医院放疗科,辽宁沈阳110001

出　　处：《中国微生态学杂志》2013年第2期147-149,共3页Chinese Journal of Microecology

基　　金：辽宁省博士启动课题(20111108)

摘　　要：目的通过在人卵巢癌细胞SKOV-3中转染NK4基因表达片段,研究NK4对人卵巢癌的治疗作用,从而验证NK4可作为卵巢癌潜在的基因治疗新策略。方法用NK4和荧光霉素表达质粒分别稳定转染人卵巢癌细胞SKOV-3。用West-ern blot检测细胞培养基中NK4蛋白表达,以及经不同培养基(SKOV-3、SKOV-3/LUC和SKOV-3/NK4培养上清)培养后SKOV-3细胞中c-Met和磷酸化-c-Met的表达。用MTT试剂盒绘制细胞生长曲线。用细胞划痕试验检测NK4对细胞转移的作用。结果在SKOV-3/NK4培养基中有NK4蛋白表达,对照(SKOV-3,SKOV-3/LUC)细胞中无表达。磷酸化-c-Met在SKOV-3/NK4培养基培养的SKOV-3细胞中被抑制,而在对照(SKOV-3,SKOV-3/LUC)细胞培养基培养的SKOV-3细胞中正常表达。c-Met表达在各组差异无统计学意义。三种细胞体外生长曲线差异无统计学意义(P>0.05)。细胞划痕试验表明SKOV-3/NK4划痕区域的细胞个数明显少于SKOV-3和SKOV-3/LUC细胞。结论 NK4蛋白在SKOV-3/NK4细胞培养基中大量分泌,NK4能抑制人卵巢癌细胞c-Met受体磷酸化,并能抑制卵巢癌细胞体外活动能力,为应用NK4作为卵巢癌基因治疗方法提供了研究基础。Objective To investigate the antitumor activity of NK4 in ovarian cancer and prove NK4 as a potential molecular therapeutic strategy against ovarian cancer.Methods NK4 cDNA was introduced into human ovarian cancer cell line SKOV-3 and the effect of NK4 after gene transfer on biological behaviors of SKOV-3 were observed.Western blot analysis was used to detect NK4 expression in the supernatants of SKOV-3,SKOV-3/LUC and SKOV-3/NK4 cells and to detect c-Met and phospho-c-Met expression in SKOV-3 cells cultured with supernatants from SKOV-3,SKOV-3/LUC and SKOV-3/NK4 cells.Cell growth curves were performed using MTT kit.Migration capabilities of the cells were evaluated in vitro using scratch wound healing assay.Results NK4 was detected in the supernatant of SKOV-3/NK4,and NK4 expressed in the supernatant of SKOV-3/NK4 inhibited the phosphorylation of c-Met in SKOV-3 cells.There was no intergroup difference among SKOV-3,SKOV-3/LUC or SKOV-3/NK4 cells in doubling time in vitro.The number of migrated cells was significantly smaller in the SKOV-3/NK4 cells than that in the control（SKOV-3 or SKOV-3/LUC） cells.Conclusion NK4 can inhibit the phosphorylation of c-Met in SKOV-3 cells and inhibit the cell migration capability of SKOV-3 cells.These findings suggest that NK4 has the potential to be a novel gene targeted therapeutic strategy to human ovarian cancer.

关 键 词：NK4 卵巢癌 基因治疗 

分 类 号：R378.992[医药卫生—病原生物学]