BRCA2基因在免疫杀伤肺癌细胞中的作用及机制研究

Role and mechanism of BRCA2 gene in immune killing of lung cancer cells

目的:研究BRCA2基因对免疫细胞杀伤肺癌的作用及分子机制。方法:siRNA敲减肺癌细胞中的BRCA2基因,qPCR及Western blot技术检测BRCA2表达;MTT及CCK-8法检测细胞生长;将外周血单核细胞与肺癌细胞共培养,酶标法检测GFP荧光强度,评估肺癌细胞杀伤效率。结果:BRCA2基因在肺癌细胞A549与H1299中表达,与肺上皮细胞BEAS-2B相比差异无统计学意义(P>0.05);当细胞A549与H1299中BRCA2基因被成功敲减后,细胞增殖率与对照组相比显著升高;外周血单核细胞对A549与H1299细胞的杀伤效率与对照组相比显著增强,ATM、RAD51、RAD50分子表达与对照组相比显著减少,而P53蛋白表达是对照组的7.2倍。结论:敲减BRCA2基因后,外周血单个核细胞对肺癌细胞的杀伤更强,干预BRCA2与单核细胞有协同抑制肺癌效应,并且调控ATM信号通路分子表达。

Objective:To study the effect and molecular mechanism of BRCA2 gene on the killing of lung cancer by immune cells.Methods:siRNA was designed to reduce BRCA2 gene in lung cancer cells;BRCA2 expression was detected by qPCR and Western blot;cell growth was detected by MTT and CCK-8 methods;peripheral blood mononuclear cells were co-cultured with lung cancer cells,and GFP fluorescence was detected by enzyme labeling method.Killing efficiency of lung cancer cells was evaluated.Results:BRCA2 gene was expressed in moderate abundance in lung cancer cells A549 and H1299,and there was no significant differ-ence compared with lung epithelial cells BEAS-2B(P>0.05).When the BRCA2 gene in A549 and H1299 cells was successfully knocked down,the cell proliferation rate was significantly increased compared with control group;the killing efficiency of peripheral blood mononuclear cells to lung cancer cells A549 and H1299 were significantly higher than that of the control group;the expression of ATM,RAD51 and RAD50 were significantly reduced,while the expression of P53 protein was 7.2 times of the control group.Con⁃clusion:After knockdown of BRCA2 gene,peripheral blood monocytes are more effective in killing lung cancer cells.Intervention of BRCA2 and monocytes can synergistically inhibit lung cancer and regulate the expression of ATM signaling pathway molecules.