白血病抑制因子受体过表达对肺癌干细胞凋亡的影响及其机制  

Effect of overexpressed leukemia inhibitory factor receptor on apoptosis of lung cancer stem cells

作　　者：姜琪 任雪松[1] 刘春华 张艳[1] 汤秦 JIANG Qi;REN Xuesong;LIU Chunhua;ZHANG Yan;TANG Qin(Sichuan Mianyang 404 Hospital, Mianyang 621000, China)

机构地区：[1]四川绵阳四0四医院,四川绵阳621000 [2]绵阳市中心医院

出　　处：《山东医药》2018年第20期1-4,共4页Shandong Medical Journal

基　　金：四川省卫生和计划生育委员会科技发展计划项目(2016PJ187)

摘　　要：目的探讨白血病抑制因子受体(LIFR)过表达对肺癌干细胞(LCSC)凋亡的影响及其机制。方法从肺癌患者的原发癌灶中分离培养原代肺癌细胞,用流式细胞术筛选出CD133+表型的LCSC。将包含LIFR的重组慢病毒过表达质粒及其阴性对照空载体质粒分别以病毒/细胞数量为18的比例感染LCSC,经2.0μg/m L的嘌呤霉素筛选,成功构建稳定表达LIFR的LCSC细胞株及其对照细胞株,分别作为观察组和对照组。采用实时荧光定量PCR法检测两组LIFR mRNA相对表达量,采用流式细胞术检测培养48 h时两组细胞凋亡率,采用Western blotting法检测两组细胞凋亡相关蛋白Bax、Bcl-2和Caspase-3相对表达量。结果观察组、对照组LIFR mRNA表达量分别为8.54±1.32、1.03±0.02,细胞凋亡率分别为(35.00±5.29)%、(11.33±1.76)%,两组比较P均<0.05。与对照组比较,观察组Bcl-2蛋白相对表达量降低(P<0.05),Bax、Caspase-3蛋白相对表达量升高(P均<0.05)。结论 LIFR过表达可促进LCSC凋亡;其机制可能与抑制Bcl-2蛋白表达,促进Bax、Caspase-3蛋白表达有关。Objective To explore the effect and mechanism of overexpressed leukemia inhibitory factor receptor（LIFR） on the apoptosis of lung cancer stem cells（LCSCs）. Methods The primary lung cancer cells were isolated and cultured from primary cancer focus of patients with lung cancer. The CD133＋phenotype of LCSCs was selected by flow cytometry. The recombinant lentiviral overexpression plasmid containing LIFR and its negative control empty vector were used to infect LCSCs at the ratio of virus/cell number = 18,and they were screened by 2 micron g/m L of purinomycin. The LCSCs with stable expression of LIFR and their control cell lines were successfully constructed,which were taken as the observation group and the control group,respectively. The relative expression of LIFR mRNA in the two groups was detected by real-time quantitative PCR. The apoptosis rate of two groups was detected by flow cytometry at 48 h. The apoptosis-related proteins Bax,Bcl-2,and Caspase-3 in the two groups were detected by Western blotting. Results The expression levels of LIFR mRNA in the observation group and control group were 8. 54 ± 1. 32 and 1. 03 ± 0. 02,and the apoptosis rates were35. 00% ± 5. 29% and 11. 33% ± 1. 76%,respectively（all P〈0. 05）. Compared with the control group,the expression of Bcl-2 protein in the observation group decreased,and the expression of Bax and Caspase-3 protein increased（all P〈0. 05）. Conclusion Overexpression of LIFR can promote the apoptosis of LCSCs by inhibiting the expression of Bcl-2 protein and promoting the expression of Bax and Caspase-3 protein.

关 键 词：肺癌 白血病抑制因子受体 肺癌干细胞 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]