HK2通过Akt1/p-Akt1上调Cdc42表达促进宫颈癌细胞迁移和侵袭

HK2 promotes cell migration and invasion by up-regulating the expression of Cdc42 through Akt1/p-Akt1 in cervical cancer cells

目的:探究己糖激酶2(HK2)通过Akt1/p-Akt1/Cdc42促进宫颈癌细胞迁移和侵袭的作用机制。方法:G418压力筛选并获取稳定表达HK2蛋白的HeLa和SiHa细胞系;Western blot和细胞免疫化学鉴定HK2蛋白在HeLa和SiHa细胞系中的表达水平;细胞划痕实验检测HK2对HeLa和SiHa体外划痕愈合能力的影响;Transwell迁移、侵袭实验检测HK2对HeLa和SiHa细胞体外迁移和侵袭能力的影响;GEPIA数据库分别分析宫颈癌中HK2的表达与Akt1、Cdc42表达的相关性;Western blot检测Akt1、p-Akt1、Cdc42蛋白在HK2过表达的HeLa细胞和SiHa细胞中的表达情况;在HK2过表达的HeLa、SiHa细胞中应用Akt1/p-Akt1抑制剂MK2206观察Cdc42的表达及细胞迁移和侵袭能力的变化。结果:成功构建HK2稳定表达的HeLa、SiHa细胞系;过表达HK2促进了HeLa、SiHa细胞的划痕愈合能力,促进了HeLa、SiHa细胞的体外迁移和侵袭;GEPIA在线数据库结果显示在宫颈癌中HK2表达与Akt1、Cdc42表达均呈正相关;过表达HK2上调了Akt1、p-Akt1、Cdc42蛋白在HeLa、SiHa细胞中的表达;在HK2过表达HeLa、SiHa细胞中,MK2206的使用抑制了HK2对Cdc42表达上调及细胞迁移和侵袭的促进作用。结论:HK2可能通过Akt1/p-Akt1途径上调Cdc42蛋白的表达促进HeLa和SiHa细胞的迁移和侵袭。

Objective:To investigate the related mechanism of HK2 on promoting cervical cancer cell migration and invasion through Akt1/p-Akt1/Cdc42.Methods:HeLa and SiHa cells with stable HK2 expression were screened.The expression of HK2 protein in HeLa and SiHa cell lines was determined by Western blot and cellular immunochemistry.Wound-healing assay,Transwell cell migration and invasion assay were used to detected the migratory and invasive potential in HK2-overexpressing HeLa and SiHa cells.The correlation between HK2 and Akt1,Cdc42 expression in cervical cancer was analyzed in GEPIA online database.The expression of Akt1,p-Akt1 and Cdc42 was detected by Western blot in HK2-overexpressing HeLa and SiHa cells.The potential of cell migration and invasion was detected in HK2-overexpressing HeLa and SiHa cells by treating with Akt1/p-Akt1 inhibitor(MK2206).Results:HK2 stably expressing HeLa and SiHa cell line was successfully constructed.HK2 enhanced cell migration and invasion in HeLa and SiHa cells.The positive correlation between HK2 and Akt1,Cdc42 expression in CESC was confirmed from the GEPIA online database.HK2 overexpression up-regulated the expression of Akt1,p-Akt and Cdc42 in HeLa and SiHa cells.The promoted cell migration and invasion in HK2-overexpressing HeLa and SiHa cells were inhibited by treating with the Akt1/p-Akt1 inhibitor MK2206.Conclusion:HK2 could up-regulating the expression of Akt1,p-Akt1 and Cdc42,further enhancing cell migration and invasion in HeLa and SiHa cells.