Journal of Shanghai Jiao Tong University (Medical Science) >

2022 , Vol. 42 >Issue 6: 742 - 750

DOI: https://doi.org/10.3969/j.issn.1674-8115.2022.06.008

Basic research

Study on the function of TRMT61A in liver cancer cell and its mechanism

  • Zhexuan HU ,
  • Xin ZHANG ,
  • Lulu WO ,
  • Jingchi LI ,
  • Jiao WANG ,
  • Cixiang ZHOU ,
  • Qian ZHAO
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  • 1.Department of Pathophysiology, Key Laboratory for Cell Differentiation and Apoptosis Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
    2.Research Units of Stress and Tumor(2019RU043), Chinese Academy of Medical Sciences, Shanghai 200025, China
ZHAO Qian, E-mail: qzhao@shsmu.edu.cn.

Received date: 2022-03-16

  Accepted date: 2022-05-25

  Online published: 2022-08-19

Supported by

National Natural Science Foundation of China(81772831)

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Abstract

Objective

·To investigate the effect of tRNA methyltransferase 61 homolog A (TRMT61A) on liver cancer cell function and its mechanism.

Methods

·The expression of TRMT61A in tumor and paired peri-tumor tissues of hepatocellular carcinoma patients was analyzed from TCGA database and the survival curves were plotted by using bioinformatics software. Stably-TRMT61A-knockdown Huh7 cells and HepG2 cells were established by using CRISPR-Cas9 system. TRMT61A protein levels in negative control group and knockdown group were detected by Western blotting. M1A methylation levels in the two groups were detected by Dot blot assay. The cell proliferation of the two groups was investigated through CCK-8 assay and colony formation assay. Flow cytometry was used to analyze cell cycle after propidium iodide (PI) staining. Cell cycle-related protein level was detected by Western blotting. Cell apoptosis was detected by using Annexin V/PI kit. Cell apoptosis-related protein level was detected by Western blotting.

Results

·TRMT61A was highly expressed in hepatocellular carcinoma samples from TCGA database through bioinformatic analysis. Survival curves showed that TRMT61A was negatively correlated with patient prognosis. TRMT61A protein level and m1A modification level were lower in TRMT61A-knockdown liver cancer cells. CCK-8 assay showed that the proliferation ability was aberrantly inhibited in Huh7 and HepG2 cells after TRMT61A knockdown. Colony formation assay showed that the number of colonies was reduced in Huh7 and HepG2 cells after TRMT61A knockdown. Mechanism investigation showed that TRMT61A-knockdown Huh7 and HepG2 cells both displayed cell cycle arrest on G0/G1 phase with elevated P21 protein level and decreased cyclin D1 protein level. Cell apoptosis rates were higher in Huh7 and HepG2 cells after TRMT61A knockdown with elevated cleaved-caspase3 protein level.

Conclusion

·TRMT61A knockdown inhibits the proliferation of liver cancer cells and induces cell apoptosis.

Key words： N1-methyladenosine (m1A) methylation; TRMT61A; hepatocellular carcinoma; cell proliferation; cell apoptosis

Cite this article

Zhexuan HU , Xin ZHANG , Lulu WO , Jingchi LI , Jiao WANG , Cixiang ZHOU , Qian ZHAO . Study on the function of TRMT61A in liver cancer cell and its mechanism[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022 , 42(6) : 742 -750 . DOI: 10.3969/j.issn.1674-8115.2022.06.008

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