m6A RNA甲基转移酶METTL3对肠癌细胞上皮间质转化及侵袭能力的影响  

The Role of METTL3 in the EMT and Invasion Ability in Colorectal Cancer Cells

作　　者：蒯榕[1] 徐莹[1] 褚以忞[1] 周璐 张海芹 李吉[1] KUAI Rong;XU Ying;CHU Yi-min;ZHOU Lu;ZHANG Hai-qin;LI Ji(Digestive Endoscopy Center,Tongren Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200336,China)

机构地区：[1]上海市同仁医院上海交通大学医学院附属同仁医院内窥镜室,上海200336

出　　处：《现代生物医学进展》2021年第21期4019-4023,4074,共6页Progress in Modern Biomedicine

基　　金：上海交通大学医工(理)交叉基金项目(ZH2018QNB24);上海市长宁区科学技术委员会一般项目(CNKW2018Y02)。

摘　　要：目的:探讨METTL3对肠癌细胞上皮间质转化及侵袭能力的影响。方法:TGFβ诱导肠癌细胞HCT116和SW480发生上皮间质转化;细胞划痕实验和Transwell-Matrigeal穿膜试验分别检测细胞转移侵袭能力;实时荧光定量PCR检测靶基因转录水平;si RNA干扰技术敲减肠癌细胞METTL3水平。结果:细胞划痕修复实验表明:TGFβ诱导组相对愈合面积与对照组相比显著增大(P<0.0001),而TGFβ+敲减METTL3联合组处理组相对愈合面积与TGFβ诱导组相比显著减少(P<0.0001)。Transwell-Matrigeal细胞穿膜试验表明:TGFβ诱导组穿膜细胞数较对照组细胞显著增多(P<0.0001),而TGFβ+敲减METTL3联合组处理组与TGFβ诱导组相比显著减少。TGFβ诱导组可显著诱导肠癌细胞发生上皮间充质转化,即增加核心转录因子Snail1和ZEB1水平,减低CDH1及上调Vimentin(VIM)水平,METTL3表达随TGFβ处理时间逐步上调。TGFβ+敲减METTL3联合组较TGFβ诱导组ZEB1和Snail1转录水平显著减低。结论:METTL3在TGFβ介导肠癌细胞EMT中起到关键作用,敲减METTL3可削弱肠癌细胞转移和侵袭能力,靶向抑制METTL3可能成为干预结直肠转移的重要分子靶点。Objective:To investigate the impact of METTL3 on EMT and invasion ability in colorectal cancer cells.Methods:TGFβwas used to induce EMT in HCT116 and SW480 cells.Wound scratch assay and Transwell-matrigeal assay were used to evaluate the invasion ability of CRC cells.qPCR was used to measure the expression of target genes.Si RNAs were used to knockdown METTL3 in CRC cells.Results:The healing area in the TGFβtreated cells was larger than control cells(P<0.0001),while TGFβplus METTL3 knockdown groups cells showed smaller healing area than TGFβtreated cells(P<0.0001).Transwell-Matrigeal showed the penetrated cells in the TGFβtreated cells was more than control cells(P<0.0001),while TGFβplus METTL3 knockdown groups cells showed less penetrated cells than TGFβtreated cells(P<0.0001).TGFβtreatment induced EMT,while expression of snail1,ZEB1,VIM and METTL3 was up-regulated and CHD1 was down-regulated.TGFβplus METTL3 knockdown groups cells showed less expression of snail1 and ZEB1 than TGFβtreated cells.Conclusion:METTL3 plays key role in the TGFβinduced EMT.Knockdown of METTL3 can impair TGFβ’s effect,which provides novel target for blocking metastasis in CRC.

关 键 词：METTL3 上皮间质转化 转移 TGFΒ 

分 类 号：R-33[医药卫生] R735.3