METTL14促进肝纤维化的机制研究

肝脏 ›› 2021, Vol. 26 ›› Issue (2): 159-161.

METTL14促进肝纤维化的机制研究

张君佩, 陈颖, 沈丹杰, 刘海玲, 田怡, 陈世耀

201100 上海复旦大学附属闵行医院消化内科(张君佩,陈颖,沈丹杰,刘海玲,田怡);复旦大学附属中山医院消化内科(陈世耀)

收稿日期:2020-07-07 出版日期:2021-02-28 发布日期:2021-03-28
通讯作者: 陈世耀,Email:syaochen@fudan.edu.cn

Mechanism of METTL14 promoting liver fibrosis

ZHANG Jun-pei¹, CHEN Yin¹, SHEN Dan-jie¹, LIU Hai-ling¹, TIAN Yi¹, CHEN Shi-yao²

1. Department of Gastroenterology, Minhang Hospital of Fudan University, Shanghai, 201100, China;
2. Department of Gastroenterology, Zhongshan Hospital of Fudan University, Shanghai, 200032,China

Received:2020-07-07 Online:2021-02-28 Published:2021-03-28
Contact: CHEN Shi-yao,Email:syaochen@fudan.edu.cn

摘要/Abstract

摘要： 目的研究RNA甲基化转移酶METTL14对肝星状细胞系增殖及活化的影响。方法 RT-qPCR检测TGF-β1处理后的肝星状细胞系中METTL14的表达量;在肝星状细胞系中过表达METTL14,采用MTT和Transwell的方法检测细胞的增殖及迁移,RT-qPCR和蛋白质印迹检测细胞中COL I和α-SMA的蛋白表达。结果 TGF-β1与肝星状细胞共培养后,METTL14的表达量上调了2.8倍左右。过表达METTL14后,肝星状细胞培养48 h的吸光度为(0.730 ± 0.047),高于对照组的(0.533 ± 0.022);同时细胞的迁移数目为(186.00 ± 42.75)亦高于对照组(97.00±7.00)差异均有统计学意义(P<0.05)。高表达METTL14细胞中COL I与α-SMA的相对表达水平(2.48 ± 0.31;3.36 ± 0.43)均显著高于对照组(1.00 ± 0.26;1.00 ± 0.24)差异均有统计学意义(P<0.05)。COL I与α-SMA的蛋白表达亦明显高于对照组。结论 METTL14促进肝星状细胞增殖、活化是肝纤维化的重要机制。

关键词: 肝星状细胞活化, 肝纤维化, METTL14, m6A修饰

Abstract: Objective To study the effect of methyltransferase?like 14 (METTL14) on the proliferation and activation of hepatic stellate cells (HSCs).Methods The expression of METTL14 in transforming growth factor beta 1 (TGF-β1) treated HSCs was detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). In HSCs with overexpression of METTL14, the abilities of proliferation and migration were analyzed by MTT assay and transwell migration assay, and the expression of collagen type I (COL I) and alphas-smooth muscle actin (α-SMA) was analyzed by RT-qPCR and Western blot. Results The expression of METTL14 in HSCs was up-regulated to about 2.8-fold after co-cultivation with TGF-β1. The absorbance of HSCs after 48 hours in culture of co-culture group (0.730±0.047) was significantly higher than that of control group (0.533±0.022, P<0.05). And the number of migrating cells (186±42.75) was also significantly larger than that of control group (97±7.00, P<0.05). RT-qPCR showed that the relative expression levels of COLI and α-SMA in cells with overexpression of METTL14 (2.48±0.31, 3.36±0.43) were significantly higher than those in control group (1.00±0.26, 1.00±0.24, both P<0.05). Western blot showed that the protein expression levels of COLI and α-SMA in co-culture group were also significantly higher than those in control group.Conclusion METTL14 promotes hepatic fibrosis by enhancing the proliferation and activation of hepatic stellate cells.

Key words: Hepatic stellate cell activation, Hepatic fibrosis, METTL14, m6A modification

张君佩, 陈颖, 沈丹杰, 刘海玲, 田怡, 陈世耀. METTL14促进肝纤维化的机制研究[J]. 肝脏, 2021, 26(2): 159-161.

ZHANG Jun-pei, CHEN Yin, SHEN Dan-jie, LIU Hai-ling, TIAN Yi, CHEN Shi-yao. Mechanism of METTL14 promoting liver fibrosis[J]. Chinese Hepatolgy, 2021, 26(2): 159-161.

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