肺鳞癌LC-42及腺癌SELS细胞中PDHA1及GBP1表达的测定

Detection of PDHA1 and GBP1 expressions in lung squamous cell carcinoma LC-42 and lung adenocarcinoma SELS cells

目的:探索肺鳞癌LC-42及肺腺癌SELS细胞中PDHA1及GBP1基因活性的相互关系。方法:肺鳞癌LC-42及肺腺癌SELS细胞分别分为CT组、siRNA CT组、PDHA1 siRNA组及GBP1 siRNA组。其中CT组为空白对照组,siRNA CT组为siRNA对照组,PDHA1 siRNA组及GBP1 siRNA组为siRNA转染试剂分别沉默PDHA1或GBP1基因,转染48 h后收取细胞。采用RT-PCR检测各组细胞PDHA1及GBP1 mRNA的表达,再用免疫细胞化学染色及Western blot分析各组细胞PDHA1及GBP1蛋白的表达。结果:RT-PCR结果显示,与CT组相比,PDHA1 siRNA组的GBP1 mRNA表达水平上调(P<0.05)。免疫细胞化学染色及Western blot结果显示,与CT组相比,PDHA1 siRNA组的GBP1蛋白表达水平上调(P<0.05)。结论:肺鳞癌LC-42及肺腺癌SELS细胞中,GBP1可能是PDHA1的下游基因,PDHA1直接或间接参与了GBP1基因的负调控。

Aim:To explore the PDHA1 and GBP1 genes activity relationship in lung squamous cell carcinoma LC-42 and lung adenocarcinoma SELS cells.Methods:Lung squamous cell carcinoma LC-42 and lung adenocarcinoma SELS cells were respectively divided into CT group,siRNA CT group,PDHA1 siRNA group and GBP1 siRNA group.CT group was blank control group,siRNA CT group was siRNA control group,PDHA1 siRNA group and GBP1 siRNA group were transfected PDHA1 siRNA or GBP1 siRNA respectively.Cells were collected after 48 hours transfection.The expression of PDHA1 and GBP1 mRNA was detected by RT-PCR,and immunocytochemistry and Western blot were used to analyze the expression of PDHA1 and GBP1 protein in each group.Results:RT-PCR showed that,compared with the CT group,the expression of GBP1 mRNA was up-regulated in PDHA1 siRNA group(P<0.05).Immunocytochemical staining and Western blot showed that,compared with the CT group,GBP1 protein expression was up-regulated in PDHA1 siRNA group(P<0.05).Conclusion:In lung squamous cell carcinoma LC-42 and lung adenocarcinoma SELS cells,GBP1 maybe a downstream gene of PDHA1,and PDHA1 is directly or indirectly involved in the negative regulation of GBP1 gene.