EphA2 siRNA转染对人卵巢癌细胞SKOV3生物学行为的影响

Influence of EphA2 siRNA Transfection on the Biological Behavior of Human Ovarian Carcinoma Cell

目的探讨促红素人肝细胞受体酪氨酸激酶A2(EphA2)干扰性小RNA(siRNA)转染人卵巢癌细胞SKOV3后,对其生物学行为的影响。方法根据EphA2序列合成一对siRNA,转染SKOV3细胞,通过荧光倒置显微镜观察转染效率和细胞形态;Western blot检测EphA2蛋白表达水平;台盼蓝染色细胞计数法绘制细胞生长曲线;集落形成实验、黏附力测定、细胞侵袭重建基底膜实验测定细胞的集落形成率、黏附和侵袭力。结果构建的EphA2siRNA转染SKOV3后,荧光倒置显微镜下可见大量荧光颗粒;Western blot检测EphA2蛋白水平表达降低(P<0.05);细胞的增殖、黏附力、侵袭力均受到抑制(P<0.05);集落形成率无明显影响(P>0.05)。结论 EphA2siRNA有效降低细胞中EphA2基因表达,抑制细胞的增殖,降低细胞的黏附力和侵袭力,从而逆转其恶性生物学行为,为肿瘤的有效干预和治疗提供新的靶点。

Objective To explore EphA2 siRNA transfection and its influence on the biological behavior of human ovarian carcinoma cells. Methods One pairs of siRNA was synthesized and transfected into the human ovarian carcinoma cell line SKOV3. Observation of the transfeetion efficiency through the fluorescence inverted microscope was followed by the evaluation of expression of EphA2 protein using Western blot. The effects of F.phA2 siRNA on proliferation of SKOV3 cells were observed by drawing cell growth curves and measuring cloning efficiency, and the adhesion and invasion of SKOV3 were detected by cell adhesion assay and Matrigel-boyden chamber method respectively. Results After successful transfection, a large number of fluorescent particles were observed under the fluorescence inverted microscope. The expression of EphA2 protein was obviously suppressed by EphA2 siRNA （P（0.05）, The cell proliferation, adhesiveness and invasiveness were significantly inhibited （P〈 0.05）. However, no changes in colony-forming efficiency were observed （P〉0.05）. Conclusion Down-regulation of EphA2 gene by EphA2 siRNA in SKOV3 cell line showed retardation of ceil growth, proliferation and partial reversion of the malignant phenotype, including the ability of adhesion and invasion. EphA2 may be a new and potent target of gene therapy in ovarian carcinoma.