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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 & AIM: Heparanase (HPA) is a β-D-glucuronidase that degrades extracellular matrix and basement membrane. HPA is localized in the placenta and immune organs of human normal tissues but is expressed at high levels in many malignancies and promotes tumor invasion and metastasis. This article discusses the heparanase gene transfection of ovarian cancer SKOV3 cells on cell growth, adhesion, invasion ability. METHODS: SKOV3 cells were transfected with plasmids carrying HPA gene (pCDNA3.0-HPA) via liposome. The expression of HPA mRNA was detected by RT-PCR. The growth curve of SKOV3 and the adhesion of cells to extracellular matrix were detected by MTT assay. The adhesion and separation of cells were detected by homogeneous adhesion assay and cell separation assay. Ovarian cancer cells SKOV3 cells invasion ability; flow cytometry cell cycle; immunohistochemical detection of ovarian cancer cells HPA protein expression. Results: Compared with the control group and the no-load group, the level of HPA mRNA in the transfected group was significantly increased (t = -128.726, -96.959, P <0.01); the S phase index in the cell cycle was significantly increased; the cell growth curve showed (T = 11.824,11.234, P <0.01); the rate of homogenous adhesion decreased (t = -14.455, -18.348, P <0.01); the cell isolation rate (Χ2 = 869.847b, 796.068b, P <0.01). The number of cells penetrating the recombinant basement membrane was significantly higher (t = 11.964,12.263, P <0.01), and the expression of HPA protein in cytoplasm increased significantly. CONCLUSION: Transfection of HPA gene can promote the growth of ovarian cancer cells and enhance the invasion and metastasis of ovarian cancer cells.