FOXP3 shRNA3对肝癌细胞趋化因子及受体CXCL12、CXCL11、CXCR4、CXCR7作用的研究  

The silencing of forkhead box P3 gene inhibited expression of chemokines CXCL12,CXCL11,CXCR4 and CXCR7 in liver cancer cells

作　　者：欧希[1] 张光涛[1] 田佩凯 陈景森[3] 徐喆[4] 谢勇[1] 王爱红[1] 刘吉奎[1] 刘晓平[1] OU Xi;ZHANG Guangtao;TIAN Peikai;CHEN Jingsen;XU Zhe;XIE Yong;WANG Aihong;LIU Jikui;LIU Xiaoping(Department of Hepatopancreatobiliary Surgery,Peking University Shenzhen Hospital,Shenzhen,518036,China;Department of Hepatobiliary Surgery,Shenzhen University General Hospital,Shenzhen,518055,China;Breast Department,Shenzhen Maternity and Child Healthcare Hospital,Shenzhen,518028,China;The Third Ward Of Liver Disease,The Third People's Hospital of Shenzhen,Shenzhen 518112,China)

机构地区：[1]北京大学深圳医院肝胆胰外科,广东深圳518036 [2]深圳大学总医院肝胆外科,广东深圳518055 [3]深圳市妇幼保健院乳腺科,广东深圳518028 [4]深圳市第三人民医院肝病三区,广东深圳518112

出　　处：《岭南现代临床外科》2018年第4期371-376,382,共7页Lingnan Modern Clinics in Surgery

基　　金：深圳市科技计划项目(JCYJ20150403091443302及JCYJ20160428164539088);深圳市医疗卫生三名工程项目(SZSM201612021);广东省科技发展专项(2017B090904010)

摘　　要：目的研究FOXP3 shRNA对肝癌细胞株SMMC-7721和MHCC-97H的趋化因子及受体CXCL12、CXCL11、CXCR4、CXCR7的影响。方法设计三种编码FOXP3 shRNA的FOXP3干扰慢病毒,sh-FOXP3-1-pGreenPuro、sh-FOXP3-2-pGreenPuro和sh-FOXP3-pgreenpuro并分别转染SMMC-7721和MHCC-97H细胞。q-PCR检测各组CXCL12、CXCL11、CXCR4、CXCR7 mRNA的表达情况;Western Blot检测各组CXCL12、CXCL11、CXCR4、CXCR7蛋白的表达情况。结果经菌落PCR和测序验证证明三个FOXP3干扰慢病毒载体构建正确;三种干扰序列中sh-FOXP3-1干扰效果最明显,因此后期实验都使用sh-FOXP3-1进行下面的实验。研究显示:(1)与对照组相比,sh-FOXP3-1组的CXCL12、CXCL11、CXCR4、CXCR7 mRNA表达明显下降,差异具有统计学意义。(2)与对照组相比,sh-FOXP3-1组的CXCL12、CXCL11、CXCR4、CXCR7蛋白表达明显下降,差异具有统计学意义。结论干扰FOXP3的表达后,能减少趋化因子CXCL12、CXCL11、CXCR4、CXCR7的表达。Objective To study the effects of forkhead box P3(FOXP3)shRNA on chemokine/chemokine receptor CXCL12、CXCL11、CXCR4、CXCR7 expression in liver cancer cells.Methods Two hepatoma cell lines SMMC-7721 and MHCC-97H cells were selected.Three FOXP3 interfering lentivirus encoding FOXP3 shRNA were designed:sh-FOXP3-1-pGreenPuro,sh-FOXP3-2-pGreenPuro and sh-FOXP3-3-pGreenPuro.SMMC-7721 and MHCC-97H cells were transfected with lentivirus vector encoding shRNA-FOXP3 respectively.q-PCR was used to detect the mRNA expression of FOXP3,CXCL12,CXCL11,CXCR4 and CXCR7 in each group.Western Blot was used to detect the expression of CXCL12,CXCL11,CXCR4 and CXCR7 at the protein levels.Results Three FOXP3 interfering lentivirus carriers were constructed accordingly,and the effect of sh-FOXP3-1 was the most obvious in reducing FOXP3 expression.The following experiments were carried out using sh-FOXP3-1.The expression of CXCL12,CXCL11,CXCR4 and CXCR7 at both mRNA and protein levels in sh-FOXP3-1 group decreased significantly compared with controls.Conclusion The silencing of FOXP3 can inhibit the expression of chemokine/chemokine receptor CXCL12,CXCL11,CXCR4 and CXCR7.

关 键 词：FOXP3 shRNA 趋化因子 趋化因子受体 肝癌细胞 

分 类 号：R681.5[医药卫生—骨科学]