抑制转移相关基因1的表达对食管癌Eca109细胞增殖和凋亡的影响  

Knockdown of MTA1 inhibits the proliferation and apoptosis of esophageal cancer cells

作　　者：于耀洋 赵佳[2] 李向楠 Yu Yaoyang;Zhao Jia;Li Xiangnan(Department of Thoracic Surgery,Zhumadian Central Hospital,Zhumadian 463000,China;Department of Thoracic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]驻马店市中心医院胸外科,463000 [2]郑州大学第一附属医院胸外科,450000

出　　处：《中华肿瘤杂志》2020年第3期197-202,共6页Chinese Journal of Oncology

摘　　要：目的探讨抑制转移相关基因1(MTA1)的表达对人食管癌Eca109细胞增殖和凋亡的影响。方法采用脂质体转染法将MTA1 siRNA转染至人食管癌Eca109细胞,同时设置转染对照组和空白对照组。通过实时荧光定量PCR法和Western blot法检测转染对Eca109细胞中MTA1 mRNA和蛋白表达的影响。采用MTT法检测各组Eca109细胞的增殖能力,采用平板克隆实验检测各组Eca109细胞的克隆形成能力,采用流式细胞术检测各组Eca109细胞的凋亡情况,采用Western blot法检测各组Eca109细胞增殖细胞核抗原(PCNA)和凋亡相关蛋白caspase-3和cleaved caspase-3蛋白的表达水平。结果人食管癌Eca109细胞转染MTA1 siRNA 48 h后,实时荧光定量PCR法检测结果显示,空白对照组、转染对照组和MTA1 siRNA组Eca109细胞中MTA1 mRNA的相对表达量分别为1.00±0.10、0.98±0.09和0.21±0.03,MTA1 siRNA组Eca109细胞中MTA1 mRNA的表达水平被抑制,与空白对照组和转染对照组比较,差异有统计学意义(P<0.05)。而空白对照组与转染对照组比较,MTA1 mRNA的表达水平差异无统计学意义(P>0.05)。Western blot法检测结果与实时荧光定量PCR法检测结果一致。MTT法检测结果显示,与空白对照组和转染对照组比较,MTA1 siRNA组Eca109细胞在48、72和96 h时的吸光度(A)值均明显降低,差异均有统计学意义(均P<0.05);而空白对照组与转染对照组48、72和96 h时的A值差异无统计学意义(均P>0.05)。平板克隆形成实验检测结果显示,空白对照组、转染对照组和MTA1 siRNA组Eca109细胞的克隆形成数分别为(58.64±6.86)个、(60.02±7.04)个和(18.10±3.16)个,差异有统计学意义(P<0.05);空白对照组与转染对照组比较,细胞克隆形成能力差异无统计学意义(P>0.05)。流式细胞术检测结果显示,空白对照组、转染对照组和MTA1 siRNA组Eca109细胞的凋亡率分别为(2.13±0.54)%、(2.27±0.61)%和(32.61±5.28)%。MTA1 siRNA组Eca109细胞的凋亡率明显升高,与�Objective To investigate the effects of metastasis associated gene 1(MTA1)expression on the proliferation and apoptosis of human esophageal cancer Eca109 cells.Methods MTA1 siRNA was transfected into human esophageal cancer Eca109 cells,and the control group and blank group were set up.The expression of MTA1 in Eca109 cells with different treatment was detected by real-time fluorescent quantitative PCR(RT-PCR)and western blot.The proliferation of Eca109 cells was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide(MTT)assay.The cloning formation ability of Eca109 cells was detected by plate cloning assay.The apoptosis of Eca109 cells were detected by flow cytometry.The expression of proliferating cell nuclear antigen(PCNA)and apoptosis-related proteins,including cleaved caspase-3 and total caspase-3 protein in Eca109 cells were detected by western blot.Results After 48 hours of transfection,RT-PCR result showed that the relative expression levels of MTA1 mRNA in Eca109 cells in the blank group,control group,and siRNA group were 1.00±0.10,0.98±0.09 and 0.21±0.03,respectively.The expression of MTA1 mRNA in siRNA group was significantly inhibited(P<0.05),while no significant difference of MTA1 mRNA expression between the blank group and the control group has been found(P>0.05).Western blot results were consistent with those of RT-PCR.MTT array results showed that,compared with the blank group and transfection group,the absorbance values of Eca109 cells in siRNA group were dramatically reduced at 48,72,and 96 h(all P<0.05).There were no significant differences of absorbance values between the blank group and the control group at 48,72,and 96 h(all P>0.05).The results of the plate colony formation test showed that the number of colony formation in the blank group and control group were 58.64±6.86 and 60.02±7.04,respectively,significantly higher than 18.10±3.16 in siRNA group(P<0.05),while there was no significant difference between the blank group and control group(P>0.05).Flow cytometry

关 键 词：食管肿瘤 转移相关基因1 细胞增殖 细胞凋亡 增殖细胞核抗原 

分 类 号：R735[医药卫生—肿瘤]