重组核心蛋白聚糖转染对HepG2细胞p21^(WAF1/CIP1)表达的影响  

Influences of recombinant decorin gene transfection on p21^(WAF1/CIP1) of HepG2 hepatoma cells

作　　者：王雅丽[1] 张玉成[1] 杜珍武[1] 张桂珍[1] 

机构地区：[1]吉林大学中日联谊医院中心实验室,吉林长春130033

出　　处：《吉林大学学报（医学版）》2009年第1期104-107,196,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科研基金资助课题(20050405-7;200705265)

摘　　要：目的:将已构建完成的分泌型核心蛋白聚糖真核表达载体转染到HepG2细胞中并检测核心蛋白聚糖及p21WAF1/CIP1的表达,探讨核心蛋白聚糖抗肿瘤作用的机制。方法:HepG2细胞分为转染pcDNA3.1-DCN载体的转染组和转染pcDNA3.1空载体的对照组,脂质体介导核心蛋白聚糖真核表达载体及质粒pcDNA3.1载体分别转染到HepG2细胞中,经G418筛选建立稳定转染的细胞株,采用RT-PCR法检测核心蛋白聚糖和p21WAF1/CIP1mRNA表达;Western blotting法检测核心蛋白聚糖和p21WAF1/CIP1蛋白质的表达;免疫组化法检测核心蛋白聚糖蛋白质的表达。结果:RT-PCR检测转染组细胞核心蛋白聚糖及p21WAF1/CIP1mRNA表达的灰度比值均高于对照组(P<0.05),Western blotting检测核心蛋白聚糖及p21WAF1/CIP1蛋白质表达的灰度比值也高于对照组(P<0.05),免疫组化检测转染组细胞核心蛋白聚糖蛋白质表达的阳性细胞计数高于对照组(P<0.05)。结论:成功建立了稳定转染核心蛋白聚糖的HepG2细胞株,并证实核心蛋白聚糖能够提高p21WAF1/CIP1mRNA和蛋白质的表达。Objective To transfect a recombinant pcDNA3.1-DCN into HepG2 cells and detect the expressions of decorin （DCN） and p21^WAF1/CIP1 so as to investigate the mechanism of tumor suppression of DCN. Methods HepG2 cells were divided into pcDNA3.1-dec-HepG2 group （transfected group） and pcDNA3.1-HepG2 group （control group） . After the recombinant pcDNA3.1-DCN and pcDNA3.1 were transfected into HepG2 hepatoma cells by liposome, the stably transfected cell lines were established using G418 screening. RT-PCR method was used to detect the expressions of DCN and p21^WAF1/CIP1 mRNA; Western blotting method was used to detect the expressions of DCN and p21^WAF1/CIP1 protein; immunohistochemistry was performed to detect the expression of DCN protein. Results Compared with control group, the expressions of DCN and p21^WAF1/CIP1 mRNA were significantly increased in transfected group by RT-PCR （P（0.05）, the expressions of DCN and p21^WAF1/CIP1 protein increased obviously in transfected group by Western blotting （P （0.05）, the expression of DCN protein increased in transfected group by immunohistochemistry （P〈0.05）. Conclusion The stably transfected HepG2 cell lines with DCN is successfully established and it is proved that DCN can increase the mRNA and protein expressions of p21^WAF1/CIP1.

关 键 词：核心蛋白聚糖 P21^WAF1/CIP1 HEPG2 转染 

分 类 号：Q786[生物学—分子生物学]