乙酰肝素酶基因转染SKOV3细胞对细胞增殖、黏附、侵袭能力的影响  

The effect of HPA gene transfection on the proliferation、adhesion、invasion of human ovarian cancer cell line SKOV3

作　　者：毛敬 辛哓燕 刘玉 李萍 

机构地区：[1]第四军医大学第一附属医院妇产科,陕西西安710032

出　　处：《中国癌症杂志》2006年第1期9-13,共5页China Oncology

基　　金：中国博士后科学基金资助项目(编号:2005037776)

摘　　要：背景与目的:乙酰肝素酶(heparanase,HPA)是能降解细胞外基质和基底膜的β-D-葡萄糖醛酸内切酶。HPA在人类正常组织局限于胎盘和免疫器官有表达,但在许多恶性肿瘤中有高水平的表达,促进了肿瘤的侵袭和转移。本文探讨乙酰肝素酶基因转染卵巢癌SKOV3细胞后对细胞生长、黏附、侵袭能力的影响。方法:以携带HPA基因的质粒(pCDNA3.0-HPA)经脂质体介导转染SKOV3细胞。RT-PCR法检测转染后HPA mRNA表达水平;MTT法检测SKOV3细胞生长曲线及细胞与细胞外基质的黏附;同质黏附试验和细胞分离试验检测细胞间的黏附和分离能力;利用transwell小室检测卵巢癌细胞SKOV3细胞的侵袭能力;流式细胞仪测细胞周期;免疫组化法检测卵巢癌细胞中HPA蛋白表达。结果:与对照组和空载组比较,转染组HPA mRNA水平上调,差异有显著性(t=-128.726,-96.959,P<0.01);细胞周期中S期指数明显增多;细胞生长曲线显示细胞生长速度增快;对细胞外基质的黏附率明显升高(t=11.824,11.234,P<0.01);同质黏附率降低(t=-14.455,-18.348,P<0.01);细胞分离率增强(χ2=869.847b,796.068b,P<0.01);穿透重组基底膜的细胞数目显著增高(t=11.964,12.263,P<0.01);细胞浆中的HPA蛋白表达显著上升。结论:转染HPA基因可以促进卵巢癌细胞的生长增值,增强卵巢癌细胞的侵袭转移能力。Background and purpose： Heparanase is an endo-β-D-glucuronindise that degrades the HS sdie chains of heparin sulphate proteoglycans in ECM, expressed in the human normal placenta and immune organ. Expression of heparanase significantly increases in many kinds of cancers, it plays a central and specific role in the regulation of tumor metastasis and invasion. This article evaluates the effect of heparanase gene on the proliferation, adhesion and invasion of human ovarian cancer cell line SKOV3 cells. Methods： Human ovarian cancer cell line SKOV3 was cultured in optimum medium and transfected with pCDNA3.0-HPA by Lipofectamine^TM2000, and cells transfected with blank vector pCDNA3.0 were used as control. The expression of HPA mRNA, and HPA protein in SKOV3 , SKOV3/neo, and SKOV3/HPA were detected by RT-PCR and immunohistoehemical staining; cell growth kinetics and the ability of invading was detected by MTT and transwell-ECM methode; the ability of adhesion and separation among same cells were evaluated by cell-adhering and separating method; Flow cytometry was used to test cell cycle. Results： RT-PCR and immunohistochemical staining showed that over-expression of HPA mRNA and protein in pCDNA3.0-HPA transfected cells in comparison with that in the parental SKOV3 cells and the cells transfected with blank vector（ P 〈 0.01）. S phase arrest was observed in SKOV3/HPA rather than in SKOV3 and SKOV3/neo cells. Cultured ovarian cancer cells transfected with HPA construct displayed more invasive, proliferative and adhesive properties compared to control group（ P 〈 0.01）. Conclusions： The overexpression of HPA mRNA and protein facilitate ovarian cancer cell growth, invasion and metastasis.

关 键 词：乙酰肝素酶 转染 卵巢癌 侵袭 培养的肿瘤细胞 

分 类 号：R73-37[医药卫生—肿瘤]