SHMT2通过上调Akt1促进人皮肤黑色素瘤细胞的迁移和侵袭

SHMT2 promote cell migration and invasion by up-regulating the expression of Akt1 in skin cutaneous melanoma cell

目的:研究丝氨酸羟甲基转移酶2(SHMT2)通过上调Akt1促进人皮肤黑色素瘤细胞迁移和侵袭的分子机制。方法:通过嘌呤霉素压力筛选获取稳定表达SHMT2蛋白的A-375和WM35细胞系;通过Western Blotting和细胞免疫化学检测SHMT2蛋白在A-375和WM35细胞系中的稳定表达;通过细胞划痕、Transwell小室实验检测SHMT2对A-375和WM35细胞体外划痕愈合、迁移和侵袭能力的影响;通过GEPIA在线数据库分析SHMT2与Akt1及上皮-间充质转换(EMT)相关因子Snai1、Snai2、ZEB2和vimentin在人皮肤黑色素瘤中表达的相关性;通过RT-PCR、Western Blotting检测Akt1、p-Akt1、Snai1、Snai2、ZEB2和vimentin在SHMT2稳定表达细胞系中的表达情况。结果:成功构建SHMT2稳定表达的A-375和WM35细胞系;过表达SHMT2可以促进A-375和WM35细胞的体外划痕愈合、迁移和侵袭;GEPIA在线数据库结果显示在人皮肤黑色素瘤中SHMT2表达与Akt1、Snai1、Snai2、ZEB2和vimentin表达呈正相关;过表达SHMT2可以上调Akt1和p-Akt1蛋白在人皮肤黑色素瘤细胞中的表达。结论:SHMT2通过上调Akt1表达促进A-375和WM35细胞的体外迁移和侵袭。

Objective:To investigate the related mechanism of SHMT2 on promoting skin cutaneous melanoma cells migration and invasion through Akt1/p-Akt1.Methods:A-375 and WM35 cells with stable SHMT2 expression were screened.Wound-healing assay,transwell cell migration and invasion assay were used to detected the migratory and invasive potential in SHMT2-overexpressing A-375 and WM35 cells.The correlation between SHMT2 and Akt1,Snai1,Snai2,ZEB2 and vimentin expression in skin cutaneous melanoma(SKCM) was analyzed in GEPIA online database.The mRNA expression of Akt1,Snai1,Snai2,ZEB2 and vimentin was detected by RT-PCR in SHMT2-overexpressing A-375 and WM35 cells.The expression of Akt1 and p-Akt1 was detected by Western Blotting in SHMT2-overexpressing A-375 and WM35 cells.Results:SHMT2 stably expressed A-375 and WM35 cell line was successfully constructed.SHMT2 enhanced cell migration and invasion in A-375 and WM35 cells.The positive correlation between SHMT2 and Akt1,Snai1,Snai2,ZEB2 and vimentin expression in SKCM was confirmed from the GEPIA online database.SHMT2 up-regulated the expression of Akt1/p-Akt in A-375 and WM35 cells.Conclusion:SHMT2 could up-regulating the expression of Akt1/p-Akt and other EMT-related factors in A-375 and WM35 cells,further enhancing cell migration and invasion.