摘要
目的:探讨低表达DJ-1对肺癌细胞增殖凋亡及PI3K/AKT通路的影响。方法:采用Western blot检测肺癌细胞系中DJ-1蛋白的表达;以脂质体法转染干扰SK-MES-1细胞中DJ-1蛋白的表达后,MTT法检测细胞的增殖变化,流式细胞仪检测细胞的凋亡情况,Western blot检测细胞中p-AKT和AKT蛋白的表达水平;将PI3K/AKT通路抑制剂LY294002处理SK-MES-1细胞48 h后,MTT法和流式细胞仪分别检测细胞的增殖和凋亡情况,Western blot检测细胞中p-AKT和AKT蛋白的表达。结果:与正常肺上皮BEAS-2B细胞相比,肺腺癌A549细胞和肺鳞癌SK-MES-1细胞中DJ-1蛋白的相对表达量均显著升高,且SK-MES-1细胞中DJ-1蛋白的表达量高于A549细胞,差异均有统计学意义(P<0.05)。转染后siRNA DJ-1组细胞中DJ-1蛋白的表达量明显低于对照组(P<0.05);与对照组相比,转染24 h后siRNA DJ-1组细胞的吸光值(OD值)变化不显著(P>0.05),而转染48 h和72 h后细胞的OD值明显降低(P<0.05);转染48 h后,与对照组相比,siRNA DJ-1组细胞的凋亡率显著升高(P<0.05),p-AKT/AKT值显著降低(P<0.05)。SK-MES-1细胞经抑制剂LY294002处理48 h后,细胞的增殖凋亡趋势与下调DJ-1表达的结果相一致。结论:DJ-1在肺癌细胞中高表达,下调其表达能够抑制细胞增殖,促进细胞凋亡,其作用机制可能与PI3K/AKT信号通路有关。
Objective:To investigate the effects of low expression of DJ-1 on proliferation,apoptosis and PI3 K/AKT pathway in lung cancer cells.Methods:The expression of DJ-1 protein in lung cancer cell lines was detected by Western and blot.The expression of DJ-1 protein was interfered by liposome transfection in SK-MES-1 cells,and MTT method was used to detect the proliferation of cells,and the apoptosis of cells was detected by flow cytometry,and the expression levels of p-AKT and AKT proteins in cells were detected by Western and blot.The PI3 K/AKT pathway inhibitor LY294002 was treated with SK-MES-1 cells 48 h later.The proliferation and apoptosis of the cells were detected by MTT and flow cytometry,and the expression of p-AKT and AKT proteins in the cells was detected by Western and blot.Results:Compared with normal lung epithelial BEAS-2 B cells,the relative expression of DJ-1 protein in lung adenocarcinoma A549 cells and lung squamous cell carcinoma SK-MES-1 cells was significantly increased,and the relative expression of DJ-1 protein in SK-MES-1 cells was higher than that in A549 cells,which difference was statistically significant(P<0.05).Compared with the control group,the absorbance value(OD) cells in siRNA DJ-1 group did not change significantly after transfection 24 h(P>0.05),but the cell OD value was decreased significantly after transfection 48 h and 72 h(P<0.05).After transfection 48 h,the apoptotic rate of cells in siRNA DJ-1 group was increased significantly(P<0.05) and the p-AKT/AKT value was decreased significantly compared with the control group( P < 0. 05). Treated with LY294002 inhibitor after 48 h,proliferation and apoptosis of SK-MES-1 cells were consistent with the downregulation of DJ-1 expression. Conclusion: DJ-1 was highly expressed in lung cancer cells,and low expression of DJ-1 can inhibit cell proliferation and promote apoptosis,which mechanism may be related to PI3 K/AKT signaling pathway.
作者
张希
谭国民
廖巾琼
李东方
胡利
李丽琼
陈小会
刘巧玲
Zhang Xi;Tan Guomin;Liao Jinqiong;Li Dongfang;Hu Li;Li Liqiong;Chen Xiaohui;Liu Qiaoling(Respiratory Medicine Affiliated Teaching Hospital of Medical University of Chongqing ( Traditional Chinese Medicine Hospital Dianjiang Chongqing) , Chongqing 408300 , China;Department of Respiratory Medicine , Department of Internal Medicine, School of Clinical Medicine, Chongqing Medical University, Chongqing 400016, China;Oncology Department , Affiliated Teaching Hospital of Medical University of Chongqing ( Traditional Chinese Medicine Hospital Dianjiang Chongqing) , Chongqing 408300 , China;Medicine of Traditional Chinese Medicine, Chongqing Medical University , Chongqing 400016 ,China)
出处
《现代肿瘤医学》
CAS
2019年第12期2060-2065,共6页
Journal of Modern Oncology
基金
重庆医科大学附属教学医院科技计划项目(编号:djkjxm2017jsyfysfyy041)
