SAMD13通过激活ERK1/2对胶质瘤细胞增殖和侵袭的影响

The effects of SAMD13 on glioma cell proliferation and invasion via activating ERK1/2

目的:检测胶质瘤组织和细胞中无菌α基序结构域蛋白13(sterile alpha motif domain-containing protein 13,SAMD13)的表达情况,并探究SAMD13表达对胶质瘤细胞增殖和侵袭的影响和调控机制。方法:使用GEPIA数据库分析胶质瘤患者癌组织中SAMD13的表达及其与预后的相关性。行RT-PCR和Western blot检测胶质瘤细胞系(U373、U87和U251)中SAMD13的表达情况。构建SAMD13过表达质粒(pIRES2-SAMD13)和SAMD13 shRNA质粒(shSAMD13),转染细胞后行Western blot、CCK-8和Transwell实验,检测过表达和沉默SAMD13基因对于U373细胞增殖和侵袭的影响,同时检查Akt1、ERK1/2和STAT3的表达和磷酸化水平。此外,在转染pIRES2-SAMD13的同时,加入ERK1/2抑制剂(U0126),通过CCK-8和Transwell实验观察其对细胞增殖和侵袭的影响。结果:胶质瘤患者癌组织中SAMD13的表达显著高于癌旁组织,并与患者不良预后密切相关。U373、U87和U251 3种胶质瘤细胞系均表达SAMD13,以U373细胞表达最为显著。在U373细胞中转染pIRES2-SAMD13和shSAMD13,可分别过表达和沉默SAMD13基因,并分别促进和抑制细胞的增殖和侵袭。此外,过表达和沉默SAMD13可分别显著增强和减弱U373细胞中ERK1/2的磷酸化,而对Akt1和STAT3的磷酸化无明显影响。U0126可显著抑制由SAMD13过表达所诱发的U373细胞增殖和侵袭,但对SAMD13的表达无明显影响。结论:胶质瘤组织和细胞中均高表达SAMD13,上调的SAMD13可通过激活ERK1/2促进胶质瘤细胞的增殖和侵袭。

Objective:To detect the expression of sterile alpha motif domain⁃containing protein 13(SAMD13)in glioma tissues and cells,and explore the effect of SAMD13 expression on the proliferation and invasion of glioma cells,as well as the underlying regulatory mechanisms.Methods:The expression and prognosis correlation of SAMD13 in glioma patients was analyzed using GEPIA database.The expression of SAMD13 in glioma cell lines(U373,U87,and U251)was examined by RT⁃PCR and Western blot.The SAMD13 overexpression plasmid(pIRES2⁃SAMD13)and SAMD13 shRNA plasmid(shSAMD13)were constructed and transfected into cells.Western blot,CCK⁃8,and Transwell assays were performed to assess the effects of SAMD13 overexpression and silencing on U373 cell proliferation and invasion,as well as to evaluate the expression and phosphorylation levels of Akt1,ERK1/2,and STAT3.Additionally,the ERK1/2 inhibitor(U0126)was added during pIRES2⁃SAMD13 transfection to further investigate cell proliferation and invasion via CCK⁃8 and Transwell assays.Results:AMD13 expression was significantly higher in glioma tumor tissues,compared with adjacent normal tissues,and it was closely associated with poor prognosis in patients.SAMD13 was expressed in all three glioma cell lines(U373,U87,and U251),with the highest expression in U373 cells. In U373 cells,transfection with pIRES2⁃SAMD13 andshSAMD13 successfully overexpressed and silenced SAMD13,respectively,which promoted and inhibited cell proliferation andinvasion,respectively. Moreover,SAMD13 overexpression significantly enhanced ERK1/2 phosphorylation,while silencing SAMD13reduced it,with no significant effects on Akt1 or STAT3 phosphorylation. The ERK1/2 inhibitor markedly suppressed the U373 cellproliferation and invasion induced by SAMD13 overexpression,but did not affect SAMD13 expression. Conclusion:SAMD13 is highlyexpressed in both glioma tissues and cells,and its upregulation promotes glioma cell proliferation and invasion by activating ERK1/2 .