敲减cGAS基因对人食管鳞癌细胞TE-1增殖、迁移和凋亡的影响  

The influence of cGAS gene silencing on proliferation,migration and apoptosis of esophageal squamous cell carcinoma TE-1 cells

作　　者：王云帆 戴东方[2] 傅聪 陈德玉[2] WANG Yun-fan;DAI Dong-fang;FU Cong;CHEN De-yu(School of Medicine,Jiangsu University, Zhenjiang Jiangsu 212013;Department of Radiotherapy, the Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001, China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]江苏大学附属医院放疗科,江苏镇江212001

出　　处：《江苏大学学报（医学版）》2020年第1期24-28,33,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81572956)

摘　　要：目的:探究环鸟苷酸腺苷酸合成酶(cyclic guanosine monophosphate-adenosine monophosphate synthase,cGAS)蛋白在食管鳞癌组织中的表达及其对人食管鳞癌TE-1细胞增殖、迁移和凋亡的影响。方法:免疫组化分析60例食管鳞癌患者癌组织与癌旁组织中cGAS的表达。荧光定量PCR和蛋白质印迹法检测正常食管上皮细胞与食管鳞癌细胞中cGAS的表达。使用慢病毒构建稳定敲减cGAS的细胞系(TE-1),荧光定量PCR和蛋白质印迹法验证敲减效率。CCK8和克隆形成实验检测cGAS敲减后TE-1细胞的增殖能力,划痕实验和Transwell实验检测细胞的迁移能力,流式细胞术检测细胞的凋亡率。结果:免疫组化结果显示cGAS在食管鳞癌组织中的表达明显高于癌旁组织;食管鳞癌细胞系TE-1、KYSE-150中cGAS的mRNA和蛋白表达明显高于食管正常上皮细胞Het-1A。荧光定量PCR和蛋白质印迹法检测结果显示,敲减cGAS后,TE-1细胞cGAS mRNA和蛋白水平的表达明显降低。CCK8、克隆形成实验、划痕实验和Transwell实验表明,敲减cGAS抑制食管鳞癌TE-1细胞增殖、迁移的能力,流式细胞分析显示敲减cGAS促进TE-1细胞凋亡。结论:敲减cGAS在食管鳞癌细胞增殖、迁移中发挥抑制作用,有效促进细胞凋亡。Objective:To investigate the expression of cyclic guanosine-monophosphate adenosine monophosphate synthase(cGAS)protein in esophageal squamous cell carcinoma,and to clarify the influence of cGAS gene silencing on proliferation,migration and apoptosis of esophageal squamous carcinoma cell line TE-1.Methods:Sixty cases of esophageal cancer and adjacent tissues were examined by immunohistochemiscal staining.The expression of cGAS in the normal esophageal epithelial cell and esophageal squamous cell was detected by fluorescence quantitative PCR and Western blotting.Lentivirus infection was used to construct stable sh-cGAS cell lines TE-1.qRT-PCR and Western blotting were used to assess cGAS knockdown efficiency.The proliferation ability of cells was detected by CCK8 assay and clone formation assay.The migration ability of cells was detected by cell scratch test and Transwell test.The apoptosis of cells was detected by flow cytometry.Results:Immunohistochemical evaluation results showed that the expression of cGAS in human ESCC was significantly higher than that of matched adjacent tissues.The expression levels of cGAS was low in normal esophageal epithelial cells Het-1A and high in esophageal squamous cell lines TE-1,KYSE-150.The cGAS expression levels of sh-cGAS groups was significantly lower than that in control group by RT-PCR and Western blotting.Compared with the control group,CCK8 assay,Colony formation assay,cell scratch test and Transwell test showed that the proliferation and migration abilities of sh-cGAS groups had significant reductions,whereas flow cytometry showed that the apoptosis of sh-cGAS groups was significant stronger.Conclusion:Knockdown of cGAS gene can inhibit the proliferation ability and migration ability,and effectively promote apoptosis.

关 键 词：食管鳞癌 环鸟苷酸腺苷酸合成酶 增殖 迁移 凋亡 

分 类 号：R735.1[医药卫生—肿瘤]