支架蛋白RACK1与蛋白激酶WEE1互作调控胃癌细胞株HGC27增殖的作用研究

目的研究活化的蛋白激酶C受体(Receptor for activated protein kinase C,RACK1)与蛋白激酶WEE1互作调控胃癌细胞株HGC27生长增殖及其作用机制。方法采用Lipofect AMINE 2000在HGC27细胞中过表达RACK1,Western blotting检测HGC27细胞中WEE1蛋白表达;免疫共沉淀验证RACK1和WEE1在HGC27细胞内存在相互作用;细胞内免疫荧光观测RACK1与WEE1在HGC27细胞中的表达定位情况。结果过表达RACK1后WEE1在胃癌细胞HGC27中的表达降低,RACK1与WEE1在HGC27细胞内存在相互作用且共定位在细胞质内。结论 RACK1与WEE1共同作用调控胃癌的发生发展过程,为RACK1成为临床上胃癌治疗的潜在靶点提供理论基础和实验依据。

The regulatory role of the interaction of scaffolding protein RACK1 with protein kinase WEE1 in the proliferation of gastric cancer cell line HGC27

Objective To study the regulatory role of the interaction of scaffolding protein RACK1 with protein kinase WEE1 in the proliferation of gastric cancer cell line HGC27 and the mechanism.Methods RACK1 was overexpressed in HGC27 cells by LipofectAMINE 2000.Western blotting was used to detect the expression of WEE1 protein in HGC27 cells.Immunoprecipitation showed that RACK1 interacted with WEE1 in HGC27 cells.Immunofluorescence was used to observe the expression and positioning of RACK1 and WEE1 in HGC27 cells.Results After expression of RACK1,the expression of WEE1 in gastric cancer cells HGC27 was decreased,and RACK1 interacted with WEE1 in HGC27 cells,and they co-localized in cytoplasm.Conclusion The interaction of RACK1 with WEE1 regulates the development and progression of gastric cancer,and it provides the theoretical basis and experimental basis for RACK1 to be a potential target for clinical treatment of gastric cancer.