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目的证实接合物蛋白CrkⅠ在卵巢癌恶性潜能中的作用。方法采用Western blot法检测卵巢癌组织、卵巢良性肿瘤组织、正常卵巢组织中Crk和Dock180蛋白的表达;用免疫沉淀法检测3种卵巢癌细胞株中Crk与Dock180蛋白的内源性结合;用小干扰RNA敲低SKOV3细胞中内源性的Crk,检测Crk表达缺失性细胞Crk蛋白表达水平、Rac1酶活性和侵袭力的变化。结果 Dock180与CrkⅠ的表达强度呈现明显的一致性,卵巢癌组织中二者的表达均显著高于卵巢良性肿瘤组织和正常卵巢组织(P<0.05);而在卵巢良性肿瘤组织与正常卵巢组织之间未发现明显差异(P>0.05)。3个卵巢癌细胞株中Dock180主要表现出与CrkⅠ的内源性结合。和对照相比,敲低Crk基因的细胞表现出侵袭能力和Rac1酶活性明显下降(P<0.05),而敲低CrkⅠ的细胞和同时敲低CrkⅠ和CrkⅡ的细胞相比无明显差异(P>0.05)。结论卵巢癌的恶性生物学行为与Crk/Dock180/Rac1信号通路相关,其中CrkⅠ在卵巢癌中扮演着主要角色。
Objective To confirm the role of CrkⅠ in the malignant potential of ovarian cancer. Methods Western blot was used to detect the expression of Crk and Dock180 in ovarian cancer tissues, benign ovarian tumor tissues and normal ovarian tissues. The endogenous binding of Crk and Dock180 protein in 3 ovarian cancer cell lines was detected by immunoprecipitation. The interference RNA knocked down the endogenous Crk in SKOV3 cells and detected the expression of Crk protein, Rac1 activity and invasiveness in Crk-deficient cells. Results The expression intensity of Dock180 and Crk Ⅰ showed obvious consistency. The expression of both in ovarian cancer tissues was significantly higher than that in benign ovarian tumors and normal ovarian tissues (P <0.05), while in ovarian benign tumors and normal ovarian tissues No significant difference was found between the two groups (P> 0.05). Dock180 in 3 ovarian cancer cell lines mainly showed endogenous binding to CrkⅠ. Compared with the control, the cells that knocked down the Crk gene showed invasion ability and Rac1 enzyme activity decreased significantly (P <0.05), but there was no significant difference between CrkⅠ knockdown cells and CrkⅠ and CrkⅡ knockdown cells (P> 0.05). Conclusions The malignant biological behavior of ovarian cancer is related to the Crk / Dock180 / Rac1 signaling pathway. Crk Ⅰ plays a major role in ovarian cancer.