HMGA1基因小分子RNAi对卵巢癌转移抑制的实验研究

Inhibitory effects of high mobility group A1 small interference RNA on metastasis of ovary carcinoma cells

目的:探讨高迁移率蛋白家族A1(high mobility group A1,HMGA1)基因小分子干扰RNA对卵巢癌转移抑制的机理。方法:RT-PCR一步法检测3种不同的卵巢癌细胞株中HMGA1、E钙黏蛋白mRNA的表达水平;设计并合成HMGA1 siRNA,转染HO8910PM细胞:半定量RT-PCR分析法观察HMGA1siRNA转染对E钙黏蛋白mRNA表达的逆转作用,及对卵巢癌细胞株体外侵袭、运动的抑制作用。结果:RT-PCR半定量分析结果显示:OVCAR-3细胞中HMGA1 mRNA表达量相对较低(0.64±0.13),而E-钙黏蛋白mRNA仅在OVCAR-3细胞中有表达;HO8910PM细胞稳定转染HMGA1 siRNA后,转染组、对照组HMGA1 mRNA表达水平分别为0.16±0.08、0.47±0.11(P<0.01),E钙黏蛋白mRNA表达水平分别为0.38±0.07、0.09±0.05(P<0.01);体外运动实验显示,跨膜细胞数转染组明显低于对照组(P<0.05);重组细胞基底膜侵袭实验显示,穿透基底膜细胞数转染组明显低于对照组(P<0.01)。结论:HMGA1基因与卵巢癌转移密切相关,HMGA1基因siRNA可上调卵巢癌细胞中E钙黏蛋白的表达,抑制卵巢癌细胞的运动和侵袭,为卵巢癌转移的基因治疗提供新的靶点。

Objective：To investigate the inhibitory effects of high mobility group A1 （HMGA1） small interference RNA （siRNA） on the migration of ovary carcinoma cell lines in vitro. Methods ：Three human ovarian carcinoma cell lines ES - 2, OVCAR - 3, HO8910PM were analyzed for the expression of HMGA1 and E - cadherin mRNA by one - step RT - PCR method. The inhibitory effects of small interference RNA of HMGA1 was detected by transwell mobility assay and matrigel invasion assay. Results：HGMA1 mRNA in OVCAR- 3 was less than in ES- 1, HO8910PM（P 〈 0.05 ） by RT - PCR. E - cadherin mRNA was only detected in OVCAR - 3. mRNA level of fibronectin was apparently down - regulated after stable transfection of siRNA HMGA1 compared to the control cells （ P 〈 0.01 ）. The mobility activity and the invasion activity of siRNA HMGA1 transfenctant cells were significantly decreased compared with control cells （ P 〈 0.01 ）. Conclusion： A close correlation of HMGA1 and metastasis and a reverse correlation of E - cadherin and HMGA1 expression exists in ovarian carcinoma cells. HMGA1 is possibly a potential target for cancer gene therapy blocking invasion and metastasis in human ovarian carcinoma.