异位过表达Pim-2诱导正常肝细胞恶变的实验研究  

Ectopic over-expression of oncogene Pim-2 induce malignant transformation of LO2

作　　者：李秀云 陈天进[2] 汪永平 刘皎 何培生[2] 

机构地区：[1]重庆垫江县中医院肝胆外科,重庆408300 [2]重庆三峡中心医院普外科,重庆404000

出　　处：《中国现代普通外科进展》2017年第4期253-260,共8页Chinese Journal of Current Advances in General Surgery

基　　金：重庆市卫生局科研资助项目(NO.2011-2-492);重庆万州区科委科研资助项目(201503023)

摘　　要：目的:研究Pim-2基因异位过表达与肝细胞性肝癌的关系。方法:将转染Pim-2基因质粒(转染组)、对照空质粒(空质粒组)转染张氏肝细胞,并设未转染组,通过MTT实验、流式细胞仪检测、Millicell小室细胞迁移实验、RT-PCR、细胞免疫组织化学及裸鼠体内接种成瘤等方法检测肝细胞在体内外增殖的变化,应用光学显微镜和透射电镜(TEM)观察细胞形态。结果:(1)转染组细胞形态发生明显变化,细胞接种48、72、96、120 h的平均OD值、细胞凋亡率、细胞迁移的平均细胞数与空质粒组相比差异有统计学意义(P<0.05),而空质粒组与未转染组比较差异无统计学意义(P>0.05);(2)转染组细胞Pim-2mRNA表达明显高于空质粒组、未转染组;转染组细胞胞质、胞核均有Pim-2蛋白表达,空质粒组、未转染组表达转弱;(3)3组细胞接种裸鼠后,成瘤组织光学显微镜及TEM均见明确肿瘤细胞形态改变。结论:Pim-2基因能够诱导张氏肝细胞恶性转变,造成细胞在形态、生长、增殖、凋亡及迁移能力的改变。Objective： To research whether ectopic over-expression of Pim-2 could cause chang-liver cell （LO2） malignant transformation, to explore the relationship between Pim-2 protein and hepatocellular carcinoma. Methods： Three groups of cells were arranged including human chang liver cell line L02 （group C）, L02 cells transfected with empty-vector （group B） and L02 cells transfected with Pim-2 gene （group A）. Pim-2 expression levels were detected. The morphology, proliferation level, apoptosis rate and migration ability of the cells were detected respectively. The cells were subcutaneously inoculated into athymic mice and the microstructures of the neoplasm were observed by optical and electron microscopy. Results： Compared with group B, Pim-2 ex- pression levels were significantly higher in group A （P〈0.05）, and their morphology had obvious ma- lignant changes. They also showed a significantly increased proliferation rate （P〈0.05） and migration capacity （P〈0.05）, as well as a significantly decreased apoptosis rate （P〈0.05）. Only the athymic mice inoculated with group A could generate neoplasm, and the morphology of the neoplasm coin- cided with that of the hepatoma. Conclusion： Both the morphological and biological changes of L02/Pim-2 cells indicate the trend of malignant transformation, which could generate hepatoma in athymic mice. Pim-2 could induce malignant transformation of human liver.

关 键 词：肝细胞肝癌 PIM-2 张氏肝细胞 裸鼠 

分 类 号：R735.7[医药卫生—肿瘤]