RAD51C表达下调对小鼠卵巢癌体内成瘤和VEGF、NRP-2表达的调控

The Effects of Down-regulating RAD51C Expression on Tumorigenesis and VEGF and NRP-2 Expression in Mouse Ovarian Cancer in Vivo

目的探讨RAD51旁系同源基因C(RAD51C)表达下调对小鼠卵巢癌体内成瘤和血管内皮生长因子(VEGF)、神经纤毛蛋白-2(NRP-2)表达调控的影响。方法使用A2780卵巢癌细胞,通过培养检测细胞中RAD51C的表达,构建3种RAD51C干扰载体(SiRNA-47、SiRNA-183和SiRNA-285),并包装成慢病毒,转染细胞,逆转录实时定量聚合酶链式反应(RT-qPCR)验证转染效果,进行稳筛,构建稳转细胞株。使用Balb/c雌性裸鼠建立细胞系来源的异体移植肿瘤模型(CDX)模型,将18只建模成功的裸鼠分为3组:A2780细胞组(Control组)、A2780细胞+空载体对照组(NC组)、A2780细胞+RAD51C干扰组(Si-RAD51C组),每组各6只。Control组注射生理盐水,NC组注射空载慢病毒50μL,Si-RAD51C组注射RAD51C干扰慢病毒50μL。观察各组成瘤情况,取肿瘤组织,采用蛋白质印迹法(WB)、免疫组织化学检测RAD51C、VEGF、NRP-2蛋白表达情况。结果RT-qPCR验证RAD51C干扰慢病毒转染效果以SiRAN-285最明显(P<0.05)。Si-RAD51C组与Control组、NC组比较瘤体的体积最小、重量也最轻,且RAD51C、NRP-2及VEGF蛋白的表达显著降低(P<0.05)。结论RAD51C干扰慢病毒可抑制小鼠A2780卵巢癌细胞肿瘤的形成,且对RAD51C、NRP-2及VEGF蛋白的表达均有抑制作用。

Objective To investigate the effects of down-regulating RAD51 paralogous gene C(RAD51C)expression on tumorigenesis and the expression of vascular endothelial growth factor(VEGF)and neuropilin-2(NRP-2)in mouse ovarian cancer in vivo.Methods 3 RAD51C interference vectors(SiRNA-47,SiRNA-183 and SiRNA-285)were constructed in A2780 ovarian cancer cells through culture to detect the expression of RAD51C in the cells,and were packaged as lentiviruses to transfect the cells.qPCR was used to verify the transfection effect,and the stable cell line was constructed.A CDX model was established using Balb/c female nude mice,and 18 nude mice were divided into 3 groups:A2780 cell group(Control group),A2780 cell+empty carrier control group(NC group),A2780 cell+RAD51C interference group(Si-RAD51C group),with 6 mice in each group.The Control group was injected with normal saline,the NC group was injected with empty lentivirus 50μL,and the Si-RAD51C group was injected with RAD51C interfering lentivirus 50μL.The tumor composition was observed,and the tumor tissues were taken,and the protein expressions of RAD51C,VEGF and NRP-2 were detected by Western blot(WB)and immunohistochemistry.Results RT-qPCR suggested that the transfection effect of RAD51C interfered with lentivirus was most pronounced with SiRAN-285(P<0.05).Compared with Control group and NC group,the tumor volume and weight of Si-RAD51C group were the smallest,and the protein expressions of RAD51C,NRP-2 and VEGF were significantly decreased(P<0.05).Conclusion RAD51C interfered with lentivirus inhibited tumorigenesis in mouse A2780 ovarian cancer cells,and inhibited the expression of RAD51C,NRP-2 and VEGF.