慢病毒介导的LKB1基因在子宫内膜癌HEC-1A细胞中的表达  

Lentivirus-mediated LKB1 gene expression in HEC-1A endometrial cancer cells

作　　者：唐乔乔[1] 宋玥[1] 龙颖[2] 张洁清[2] 宋红林[2] 赵冰冰[2] 

机构地区：[1]广西医科大学研究生学院 [2]广西医科大学附属肿瘤医院妇瘤科,南宁530021

出　　处：《中国癌症防治杂志》2014年第2期133-138,共6页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：广西研究生教育创新计划项目(1059820111002D29)

摘　　要：目的探讨慢病毒系统介导的LKB1基因在子宫内膜癌HEC-1A细胞中的过表达,为进一步研究LKB1基因在子宫内膜癌的作用机制奠定基础。方法以PCR扩增LKB1克隆质粒获得全长cDNA,将LKB1 cDNA链接到慢病毒载体pWPI,构建慢病毒表达质粒LKB1/pWPI。通过与包装质粒pCMV-Dr8.74和pMD2.G共转染293T细胞进行病毒包装,用包装成功后的病毒液感染子宫内膜癌HEC-1A细胞,以荧光定量PCR、Western blot法检测HEC-1A细胞中LKB1的相对表达量。结果成功扩增LKB1全长cDNA和构建LKB1重组慢病毒表达载体LKB1/pWPI。转染包装293T细胞后能产生慢病毒颗粒并能有效感染靶细胞HEC-1A。转染后HEC-1A-LKB1-pWPI细胞中LKB1的表达率明显高于亲本细胞和空白对照细胞(P<0.01)。结论成功构建携带LKB1基因的慢病毒表达载体,包装病毒后能有效地感染子宫内膜癌HEC-1A细胞,为进一步探讨LKB1基因在子宫内膜癌中的生物学效应奠定了基础。Objective To investigate lentivirus-mediated LKB1 gene overexpression in HEC-1A endometrial cancer cells as a basis for future research. Methods The LKB1 gene was subcloned by RT-PCR from a cDNA plasmid into a lentiviral pWPI expression vector,generating the plasmid LKB1/pWPI.This plasmid was co-transfected into 293T cells together with the packaging plasmid pCMV-Dr8.74 and pMD2.G.The recombinant virus was used to infect HEC-1A cells,and LKB1 expression was measured using real-time quantitative PCR （qRT-PCR） and Western blotting. Results We succeeded in constructing the recombinant plasmid LKB1/pWPI and packaging it into lentiviral particles in 293T cells.This virus infected HEC-1A cells efficiently,leading to higher LKB1 expression than in the parental cells or untransfected controls （P〈0.01）. Conclusion The LKB1 gene has been incorporated into a lentiviral expression vector,allowing studies of its effects on the development of endometrial cancer.

关 键 词：子宫肿瘤 子宫内膜癌 LKB1 慢病毒 

分 类 号：R737.33[医药卫生—肿瘤]