摘要
目的探讨甲基转移酶样3(METTL3)是否介导Kallikrein相关肽酶6(KLK6)的N6-甲基腺苷(m6A)修饰促进胃癌细胞的细胞活力。方法通过m6A抗体纯化胃癌细胞MGC⁃803和正常胃细胞GES⁃1中的m6A修饰的mRNA,通过实时荧光定量PCR检测这些mRNA中KLK6的相对表达量。过表达或敲低METTL3后纯化m6A修饰的mRNA,通过实时荧光定量PCR检测这些mRNA中KLK6的相对表达量。通过体外转录和METTL3体外催化KLK6的mRNA的m6A修饰。在METTL3敲低的胃癌细胞MGC⁃803中转染体外合成的m6A修饰的KLK6的mRNA,通过MTT检测胃癌细胞MGC⁃803的细胞活力。结果胃癌细胞MGC⁃803中mRNA的m6A修饰多于正常胃细胞GSE⁃1,并且胃癌细胞MGC⁃803中KLK6 mRNA的m6A修饰多于正常胃细胞GSE⁃1(P<0.05)。过表达MET⁃TL3后,胃癌细胞MGC⁃803中KLK6 mRNA的m6A修饰多于正常胃细胞GSE⁃1(P<0.05)。敲低METTL3后,胃癌细胞MGC⁃803中KLK6 mRNA的m6A修饰少于正常胃细胞GSE⁃1(P<0.05)。敲低METTL3并转染体外合成的m6A修饰的KLK6的mRNA后,胃癌细胞MG⁃803的细胞活力上升(P<0.05)。结论胃癌细胞MGC⁃803中,METTL3通过对KLK6 mRNA进行m6A修饰促进细胞活力。
Objective To investigate whether METTL3 mediates KLK6 m6 A modification to promote the cell viability of gastric cancer cells.Methods The m6 a-modified mrnas in gastric cancer cells MGC-803 and normal gastric cells ges-1 were purified by m6 A antibody,and the relative expressions of KLK6 in these mrnas were detected by real-time fluorescence quantitative PCR.After overexpression or knocking down METTL3,m6 a-modified mrnas were purified,and the relative expression of KLK6 in these mrnas was detected by real-time fluorescence quantitative PCR.In vitro transcription and METTL3 catalyzed m6 A modification of KLK6 mRNA.The m6 a-modified KLK6 mRNA was transfected into the METTL3-knockdown gastric cancer cell MGC-803,and the cell activity of gastric cancer cell MGC-803 was detected by MTT.Results The m6 A mRNA of gastric cancer cell MGC-803 was more modified than that of normal gastric cancer cell gse-1,and m6 A mRNA of KLK6 in gastric cancer cell MGC-803 was more modified than that of normal gastric cancer cell GES-1(P<0.05).After overexpression of METTL3,the m6 A of KLK6 mRNA in gastric cancer cell MGC-803 was more modified than that in normal gastric cell GES-1(P<0.05).After knocking down METTL3,the m6 A modification of KLK6 m RNA in gastric cancer cell MGC-803 was less than that of normal gastric cell GES-1(P<0.05).After knockdown of METTL3 and transfection with m6 a-modified KLK6 mRNA synthesized in vitro,the cell activity of gastric cancer cell MGC-803 increased(P<0.05).Conclusion In gastric cancer cell MGC-803,METTL3 promoted cell activity by m6 A modification of KLK6 mRNA.
作者
赵飞
李爱丽
冯运章
张学强
张伟
刘小慧
霍浩然
ZHAO Fei;LI Ai⁃li;FENG Yun⁃cheng;ZHANG Xue⁃qiang;ZHANG Wei;LIU Xiao⁃hui;HUO Hao⁃ran(Department of Gastrointestinal Sur⁃gery,Handan Central Hospital,Handan,Hebei Province 056001,China)
出处
《解剖学研究》
CAS
2020年第6期515-519,共5页
Anatomy Research
基金
河北省卫健委科技计划项目(20150454)
