分化抑制因子1通过诱导上皮间质转化促进肝癌HepG2细胞的侵袭性  

Overexpression of Id1 promotes the invasiveness of HepG2 cells by inducing epithelial to mesenchymal transition

作　　者：高璐[1] 吕刚[2] 孙凯[3] 蒋国成[1] 张长松[4] 李蓉[1] 卫立辛[1] 

机构地区：[1]第二军医大学附属东方肝胆外科医院肿瘤免疫与基因治疗研究中心,上海医学硕士研究生200438 [2]第二军医大学附属东方肝胆外科医院肝外科,上海200438 [3]交通大学医学院仁济医院中心实验室,上海200127 [4]苏州大学附属肿瘤医院临床肿瘤实验室,常州213001

出　　处：《医学研究生学报》2013年第12期1254-1257,共4页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81030041)

摘　　要：目的分化抑制因子1(inhibitor of differentiation-1,Id-1)作为一种新的癌基因,其过表达可以影响肿瘤的发生发展。文中探讨肝癌HepG2细胞中高表达Id1对E-钙黏蛋白(E-cadherin)与波形蛋白(Vimentin)及在HepG2细胞中对迁移、侵袭能力的影响。方法将已构建好的PIRES-EGFP-Id1质粒转染于肝癌HepG2细胞,采用Western blot方法检测转染48h后细胞中Id1蛋白的表达;实时定量PCR和免疫荧光方法检测E-cadherin和Vimentin的表达;利用细胞划痕实验和Transwell侵袭实验检测细胞迁移和侵袭能力的变化。结果转染组Id1蛋白表达明显上调,与空白对照组相比差异有统计学意义(P<0.05);同时进行实时定量PCR和免疫荧光检测,发现转染组E-cadherin表达下降,而Vimentin表达明显上调(P<0.05);划痕实验显示高表达Id1的HepG2组48h细胞生长迁移能力明显增强(P<0.05);侵袭实验发现转染Id1质粒组细胞穿膜数为(32.00±4.04)个,明显多于转染的空质粒组(10.67±3.84)个和空白对照组(9.67±2.72)个,差异有统计学意义(P<0.05)。结论 Id1高表达可以诱导HepG2细胞上皮向间质转化,提高肝癌HepG2细胞的迁移和侵袭能力,可通过调节E-cadherin和Vimentin发挥作用。Objective The inhibitor of differentiation-1 （ Id-1 ） is a new cancer gene and its overexpression may be associated with the occurrence and development of cancer. This study was to explore the effects of the overexpression of Id-1 in human hepatocyte cell line HepG2 on the expressions of E-eadherin and vimentin and the migration and invasiveness of HepG2 cells. Methods PIRES-EGFP-Idl plasmids were constructed and transfected into HepG2 cells. At 48 hours after transfection, the expression of the Idl protein in the HepG2 cells was determined by Western blot, and those of E-eadherin and vimentin detected by realtime PCR and immunofluoreseence staining. Changes in the migration and invasiveness of the HepG2 cells were observed by wound- healing and Transwell assay. Results Compared with the control group, the expression of the Idl protein in the HepG2 cells was significantly increased （ P 〈 0.05 ） , that of E-cadhefin down-regulated, and that of vimentin up-regulated （ P 〈 0.05 ） 48 hours after transfection. The migration capability of in the HepG2 cells with highly expressed Idl was markedly increased at 48 hours （ P 〈 0.05）. The average number of cells per field was remarkably higher in the PIRES-EGFP-Idl group （32.00 ± 4.04） than in the PIRES-EGFP group （ 10.67 ± 3.84） and the control group （9.67 ± 2.72） （ P 〈 0.05 ）. Conclusion The overexpression of Idl can induce epithelial to mesenchymal transition and promote the migration and invasiveness of HepG2 cells by inhibiting E-cadherin and up-regulating vimentin.

关 键 词：分化抑制因子1 上皮间质转化 肝癌 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]