沉默载脂蛋白B-mRNA编辑酶复合物3B基因对人胰腺癌SW1990细胞的影响  

Effect of silencing apolipoprotein B mRNA editing enzyme complex 3B gene on human pancreatic cancer SW1990 cells

作　　者：陈雪姣[1] 李昊[2] 张宏伟[2] 仓顺东[1] 唐强[2] Chen Xuejiao;Li Hao;Zhang Hongwei;Cang Shundong;Tang Qiang(Department of Oncology,People’s Hospital of Henan Province,Zhengzhou 450003,China;Department of Hepatobiliary Pancreatic Surgery,People’s Hospital of Henan Province,Zhengzhou 450003,China)

机构地区：[1]河南省人民医院肿瘤内科,郑州450003 [2]河南省人民医院肝胆胰腺外科,郑州450003

出　　处：《中华实验外科杂志》2020年第5期874-876,共3页Chinese Journal of Experimental Surgery

基　　金：河南省科技攻关项目(202102310444)。

摘　　要：目的:观察沉默载脂蛋白B-mRNA编辑酶复合物3B(APOBEC3B)基因对胰腺癌细胞PATU8988(购自美国典型菌种保藏中心)细胞周期、增殖及侵袭能力的影响。方法:通过构建重组靶向干扰APOBEC3B基因的短发卡RNA(shRNA)慢病毒表达质粒PGC-shRNA-APOBEC3B沉默APOBEC3B基因。分为空白对照组、阴性对照组及转染组。实时定量反转录聚合酶链反应(RT-qPCR)、蛋白质印迹法(Western blot)检测各组细胞APOBEC3B基因和蛋白的表达变化;流式细胞仪检测细胞周期的变化;平板克隆实验检测细胞增殖;Transwell侵袭实验检测细胞侵袭力。组间比较采用单因素方差分析和LSD- t检验。 结果:APOBEC3B基因和蛋白表达空白对照组为2.28±0.27和3.51±0.32,阴性对照组为2.32±0.21和3.33±0.29,转染组(0.26±0.09和0.31±0.12)明显低于上述两组( t=6.230、5.640, P<0.01);细胞周期结果显示G 0/G 1期与S期细胞空白对照组为(55.16±4.19)%、(31.21±1.92)%,阴性对照组为(52.35±3.53)%、(32.17±4.21)%,转染组为(75.12±5.34)%、(13.55±2.01)%,细胞周期阻止于G 0/G 1期( t=5.230, P<0.01),S期细胞数减少( t=5.830, P<0.01);平板克隆实验显示转染组细胞克隆形成数[(135.77±12.14)个]明显低于空白对照组和阴性对照组[(254.00±12.31)、(247.00±14.51)个, t=11.210, P<0.01];穿膜细胞数转染组[(92.41±18.42)个]明显低于空白对照组和阴性对照组[(215.00±23.83)、(227.00±21.25)个, t=6.480, P<0.01]。 结论:APOBEC3B基因沉默显著抑制SW1990细胞的增殖,减少S期细胞,降低细胞侵袭能力。Objective To observe the effect of short hairpin RNA(shRNA)silencing apolipoprotein B mRNA editing enzyme complex 3B(APOBEC3B)gene on cell cycle,proliferation and invasion ability of pancreatic cancer SW1990 cells.Methods The shRNA was constructed to silence APOBEC3B gene of SW1990 cells.The WW1990 cells were divided into blank control group,negative control group and transfection group.The real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the changes of APOBEC3B gene and protein expression in each group.Flow cytometry was used to detect the change of cell cycle.The plate cloning experiment was done to detect cell proliferation.SPSS 22.0 software was used to analyze the data.Results The APOBEC3B gene and protein expression in the blank control group and negative control group was 2.28±0.27,3.51±0.32;2.32±0.21,3.33±0.29,which was significantly higher than in the transfection group(0.26±0.09,0.31±0.12)(t=6.230,5.640,P<0.01).Cell cycle results showed the proportion of G0/G1 and S-phase cells in the blank control group and negative control group was(55.16±4.19)%,(31.21±1.92)%;(52.35±3.53)%,(32.17±4.21)%,and that in the transfection group was(75.12±5.34)%and(13.55±2.01)%.In the transfection group,the cell cycle was arrested in the G0/G1 phase(t=5.230,P<0.01),and the number of cells in the S-phase decreased(t=5.830,P<0.01).The plate cloning experiments showed that the number of colonies formed in the transfection group was(135.77±12.14)cells,which was significantly less than that in the blank control group and negative control group[(254.00±12.31),(247.00±14.51)cells,t=11.210,P<0.01].The number of transmembrane cells in the transfection group was(92.41±18.42)cells,which was significantly less than in the blank control group and negative control group[(215.00±23.83),(227.00±21.25)cells,t=6.480,P<0.01].Conclusion Silencing APOBEC3B gene can significantly inhibit the proliferation of SW1990 cells,reduce S-phase cells,and decrease cell

关 键 词：载脂蛋白B-mRNA编辑酶复合物3B 胰腺癌 细胞周期 脱噬作用 侵袭 

分 类 号：R735.9[医药卫生—肿瘤]