BATF2真核表达载体的构建及其对肝癌细胞Bel-7402增殖的影响  

Construction of BATF2 Eukaryotic Expression Vector and Its Effect on the Proliferation of Bel-7402 Cells

作　　者：张畅 周浮杨 郑宇航 魏超越 王璎珞 孙艳[2] ZHANG Chang;ZHOU Fuyang;ZHENG Yuhang;WEI Chaoyue;WANG Yingluo;SUN Yan(School of Basic Medical Science,Qiqihar Medical University,Qiqihar 161000,China;不详)

机构地区：[1]齐齐哈尔医学院基础医学院,黑龙江齐齐哈尔161000 [2]齐齐哈尔医学院医学技术学院

出　　处：《中国医学创新》2021年第19期27-30,共4页Medical Innovation of China

基　　金：黑龙江省大学生创新创业训练计划项目(201911230002)。

摘　　要：目的:构建人BATF2真核表达载体,并验证其过表达对肝癌Bel-7402细胞增殖的影响。方法:利用Trizol法提取正常人外周血单核细胞RNA并将其反转录成cDNA,再以cDNA为模板扩增出BATF2基因序列,再将其连接到pcDNA3.1真核表达载体上,采用酶切和测序的方法对重组质粒进行鉴定;用脂质体转染的方法将重组质粒转染Bel-7402细胞后,Western blot检测BATF2蛋白表达变化,MTT法检测对细胞增殖的影响。结果:双酶切和DNA测序结果验证了pcDNA3.1-BATF2真核表达载体构建成功;pc-DNA3.1-BATF2组BATF2蛋白表达量高于空白对照组与pcDNA3.1组(P<0.05)。转染48、72、96 h后,pc-DNA3.1-BATF2组Bel-7402细胞增殖的OD值均低于空白对照组和pcDNA3.1组(P<0.05)。结论:成功构建pcDNA3.1-BATF2真核表达载体并转染Bel-7402细胞后BATF2蛋白表达量增加,且过表达BATF2能够显著抑制肝癌细胞Bel-7402细胞增殖能力。Objective:To construct the eukaryotic expression vector of human BATF2 and verify the effect of its overexpression on the proliferation of hepatoma Bel-7402 cells.Method:The RNA of normal human peripheral blood mononuclear cells was extracted by Trizol method and reverse transcribed into cDNA.The BATF2 gene was amplified by the cDNA template,and then linked to pcDNA3.1 eukaryotic expression vector.The recombinant plasmid was identified by enzyme digestion and sequencing.The recombinant plasmid was transfected into Bel-7402 cells by the method of liposome transfection.After the recombinant plasmid was transfected into Bel-7402 cells by liposome transfection,the expression of BATF2 protein was detected by Western blot and the effect on cell proliferation was detected by MTT method.Result:The results of double enzyme digestion and DNA sequencing confirmed that the eukaryotic expression vector pcDNA3.1-BATF2 was successfully constructed.The protein expression of BATF2 in pc-DNA3.1-BATF2 group was higher than those in blank control group and PCDNA3.1 group(P<0.05).After transfection for 48,72 and 96 h,the OD values of Bel-7402 cell proliferation in pcDNA3.1-BATF2 group were lower than those in blank control group and pcDNA3.1 group(P<0.05).Conclusion:pcDNA3.1-BATF2 eukaryotic expression vector is successfully constructed and transfected into Bel-7402 cells to increase the expression of BATF2 protein,and the overexpression of BATF2 can significantly inhibit the proliferation ability of Bel-7402 cells.

关 键 词：BATF2 真核表达载体 BEL-7402细胞 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤]