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.
作者
蒯榕
徐莹
褚以忞
周璐
张海芹
李吉
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)
出处
《现代生物医学进展》
CAS
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)。
