GPC3抑制肝癌细胞增殖的机制与生长因子IGF2、BMP4之间的关系  

Association between mechanism that GPC3 suppresses growth of hepatoma carcinoma cells and growth factors IGF2、BMP4

作　　者：王瑜[1] 林山[2] 王冰[1] 姚和祥[1] 王烈[1] 

机构地区：[1]福州总医院南京军区普通外科研究所 [2]福建省莆田学院附属医院普通外科

出　　处：《福州总医院学报》2010年第1期21-23,共3页Journal of Fuzhou General Hospital

基　　金：南京军区"十五"医药卫生科研基金(02MA009)资助

摘　　要：目的:应用构建完成的GPC3增强型绿色荧光蛋白真核表达载体,研究GPC3对SK-Hep-1肝癌细胞增殖的影响及生长因子IGF2、BMP4与GPC3抑制肝癌细胞增殖之间的关系。方法:pEGFP-N2-GPC3增强型绿色荧光蛋白真核表达载体经脂质体LipofectamineTM2000介导转染SK-Hep-1肝癌细胞后,通过G418(600ug/ml)筛选出抗性克隆,应用逆转录-聚合酶链反应(RT-PCR)检测GPC3mRNA在真核细胞中的表达,Western-blot检测GPC3蛋白在真核细胞中的表达,确定GPC3已经成功转入细胞后,采用MTT法研究GPC3对SK-Hep-1肝癌细胞增殖的影响及生长因子IGF2、BMP4与GPC3抑制肝癌细胞增殖之间的关系。结果:荧光显微镜下可见转染的真核细胞胞膜区发出强绿色荧光,RT-PCR及Western-blot表明GPC3在真核细胞中成功表达,基因转染组与空白对照组、空质粒转染组相比增殖明显减慢,有显著性意义(P<0.001),GPC3对生长因子IGF2、BMP4促肝癌细胞增殖无影响(P>0.05)。结论:成功筛选稳定表达GPC3蛋白的SK-Hep-1肝癌细胞系;GPC3抑制肝癌细胞增殖;GPC3与生长因子IGF2、BMP4信号通路可能无联系。Objective： SK - Hep- 1 cells were transfected with green fluorescent protein eukaryotic expression vector pEGFP- N2 GPC3, and its role on proliferation of SK - Hep- 1 hepatoma carcinoma cells was studied and effects of GPC3 gene on the growth promotion of growth factors in SK - Hep - 1 cells were analyzed. Method： An eukaryotit expression vector for GPC3 （pEGFP- N2 - GPC3） was transfected into SK - Hep- 1 human hepatoma carcinoma cells by LipofectamineTM2000, and ce - lls that could express GPC3 stably were screened out by G418 （600ug/ml） . The mRNA expression of GPC3 was detected by RT - PCR. The protein expression of GPC3 was detected by Western - blot. Effects of GPC3 gene on the growth velocity in SK - Hep - 1 cells and GPC3 gene on the growth promotion of growth factors in SK - Hep - 1 cells were detecced by MTT. Results： The green fluorescence could be seen in transfected SK -- Hep - 1 cells membrane field under fluorescence microsco - pc. RT PCR identified that GPC3mRNA was expressed in SK - Hep - 1 cells. Western- blot identified that GPC3 protein was expressed in SK - Hep- 1 cells successfully. Forced expression of GPC3 suppressed the growth of SK- Hep- 1 cells （P〈 0. 001 ） Growth effects of IGF2 and BMP4 were not altered by GPC3. Conclusion： SK - Hep- 1 cells that could expressed GPC3 p - rotein stably were screened out successfully by O418 （600ug/ml） . Forced expression of GPC3 suppressed the growth of SK Hep- 1 cells. Forced expression of GPC3 may not modulate IGF2. BMP4 signaling.

关 键 词：GPC3基因 真核表达载体 SK—Hep-1 绿色荧光蛋白 生长因子 

分 类 号：R735.7[医药卫生—肿瘤]