目的:探讨活化型c-Src对整合素αvβ3相关肿瘤行为的影响及其分子机制,为寻找治疗肿瘤的分子靶点提供思路。方法:通过定点突变的方法构建活化型c-Src真核表达载体,应用脂质体法转染至表达有整合素αvβ3的中国仓鼠卵巢(CHO)细胞,即CHO-αvβ3细胞中建立稳定细胞株。通过流式细胞术、蛋白印迹法检测细胞中目的蛋白的表达,检测稳定细胞株在固相化αvβ3配体玻连蛋白(Vn)上的黏附、伸展功能,利用人工重组基底膜侵袭小室,观察细胞株的迁移能力,并通过蛋白免疫印迹法检测细胞中整合素β3酪氨酸磷酸化的水平。结果:流式细胞术和蛋白印迹法检测结果均证实,CHO-αvβ3中成功表达活化型c-Src。活化型c-Src对CHO-αvβ3细胞在固相化Vn上的黏附、伸展能力无明显影响,但增强了细胞迁移能力及整合素β3的磷酸化水平。结论:活化型c-Src参与αvβ3相关肿瘤行为的调节,可能与增强整合素β3磷酸化水平相关,为进一步深入了解相关机制奠定基础。 Objective: To investigate the effect of activated c-Src on integrin αvβ3-related tumor and its molecular mechanism, and to provide ideas for finding molecular targets for tumor therapy. Methods: The activated c-Src eukaryotic expression vector was constructed by site-directed mutagenesis and then transfected into Chinese hamster ovary (CHO) cells expressing integrin αvβ3 by lipofectamine to establish a stable cell line CHO-αvβ3 . The expression of the target protein in the cells was detected by flow cytometry and Western blotting, and the adhesion and extension of the stable cell lines on the immobilized αvβ3 ligand vitronectin (Vn) were detected. Invasive cells were observed by artificial recombinant basement membrane Cell lines and the level of integrin β3 tyrosine phosphorylation in the cells was detected by Western blotting. Results: The results of flow cytometry and Western blotting confirmed that activated c-Src was successfully expressed in CHO-αvβ3. Activated c-Src had no significant effect on the adhesion and spreading ability of CHO-αvβ3 cells on the immobilized Vn, but enhanced the cell migration ability and the level of integrin β3 phosphorylation. CONCLUSION: Activation of c-Src in the regulation of αvβ3-related tumor behavior may be related to the enhancement of integrin β3 phosphorylation, which may lay a foundation for further understanding of the related mechanisms.