摘要
目的:肝细胞癌中高表达磷脂酰肌醇蛋白聚糖-3基因(glypican-3,GPC3),而在非肿瘤肝组织、肝细胞腺瘤、胆管癌、肝内胆管细胞癌、胆囊癌等细胞中低表达甚至不表达;本研究利用携带GPC3重组真核表达载体,探讨GPC3基因对SK-Hep-1肝癌细胞增殖、黏附和侵袭能力的影响。方法:将pEGFP-N2-GPC3通过脂质体方法转染人肝癌细胞SK-Hep-1。RT-PCR检测GPC3-SK-Hep-1细胞中GPC3mRNA的表达;MTT法检测SK-Hep-1细胞的增殖并计算黏附率;Transwell小室实验检测SK-Hep-1肝癌细胞的迁移能力和侵袭能力。结果:pEGFP-N2-GPC3成功转染SK-Hep-1细胞,转染后GPC3-Hep-1细胞明显表达GPC3mRNA。GPC3转染能显著抑制肝癌细胞SK-Hep-1的增殖(P<0.01);GPC3转染细胞的黏附能力较对照细胞显著下降[(10.21±0.62)%vs(15.51±0.95)%,P<0.01];GPC3转染细胞的迁徙和侵袭能力较对照细胞明显增强[(131.7±7.44)vs(69.6±5.25),P<0.01;(220±12.8)vs(130±8.2),P<0.01]。结论:GPC3基因显著抑制肝癌细胞的增殖和黏附能力,但显著增强后者的迁移和侵袭能力。
Objective： To study the influence of GPC3 gene on the proliferation, adhesion and invasion of hepatoma cell line SK-Hep-1. Methods： SK-Hep-1 cells were transfected with pEGFP-N2-GPC3 using Lipofectamine2000. RT-PCR was used to examine the GPC3mRNA expression in GPC3-SK-Hep-1 cells. MTT assay was used to examine the proliferation and calculate the adhesion rate of SK-Hep-1 cells. Transwell system was used to assess the migration and invasion of the cells. Results： SK-Hep-1 cells were successfully transfected with pEGFP-N2-GPC palsmid. GPC3 mRNA was detected in SK-Hep-1 cells. Transfeetion with CPC3 significantly suppressed the growth of SK-Hep-1 cells （P 〈 0.01 ）. The adhesion ability of GPC3-transfected cells was significantly decreased compared with control group（ [ 10.21 ±0.62 ] % vs [ 15. 51 ± 0. 95 ] % , P 〈 0. 01 1. Transfection with GPC3 significantly enhanced migration and invasion capacity （ [ 131.7 ± 7.44 ] vs [ 69.6± 5.25 ] ,P 〈 0.01 ; [ 220± 12.8 ] vs [ 130±8.2 ], P 〈 0.01 ]. Conclusion： Transfection with GPC3 gene can greatly inhibit the proliferation and adhesion of hepatoma SK-Hep-1 cells, but can enhance their migration and invasiveness. The present study provides an experimental basis for studying the invasion mechanism of liver cancer.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2008年第5期474-477,483,共5页
Chinese Journal of Cancer Biotherapy
基金
福建省青年科技人才创新项目(No.2005J074)~~
