DNA methyltransferase-1 regulates SHTN1 expression in gliomas through DNA methylation
Objective To investigate the mechanisms regulating the biological function of DNA methyltransferase-1(DNMT1)in human glioma cells.Methods The bioinformatics analysis of SHTN1 was carried out using glioma-related mRNA-seq data and single-cell sequencing data in chinese glioma ge-nome atlas(CGGA),the cancer genome atlas(TCGA)and gene expression omnibus(GEO)databases.SHTN1(Shootin 1)mRNA expression levels were detected by real-time quantitative polymerase chain reac-tion to verify the correlation between DNMT1 and SHTN1 expression in human glioma cell lines with stable low-expression of DNMT1 established by lentiviral transfection and control groups named DNMT-NC(U-87,and U-251 cell lines were purchased from Shanghai Genechem Co.,Ltd.).BSAS sequencing vali-dated the regulation for methylation level of SHTN1 by DNMT1.The Kruskall-Wallis test was used to com-pare the methylation levels of each CpG site.Results The survival of glioma patients in the SHTN1 high expression group was longer than that of the low expression group(P＜0.05).The expression of SHTN1 in glioblastoma(GBM)was significantly lower than in normal brain tissue,with differences in expression ob-served across different glioma WHO grades and pathological subtypes.The mRNA expression levels of SHTN1 in the DNMT1 knockdown group in vitro experiments were significantly higher than those in the con-trol group(The expression of SHTN1 in the knockdown group of U251 was 1.7 times higher than that in the control group,P＜0.01;The expression of SHTN1 in the knockdown group of U87 was 1.4 times higher than that in the control group,P＜0.05).BSAS sequencing verified the down-regulation of methylation levels of the three CpG sites of SHTN1 in the DNMT1 low-expression group(Kruskall-Wallis P＜0.05).Conclusion The expression of SHTN1 was significantly down-regulated in high-grade gliomas,and the short-term survival of glioma patients with low expression of SHTN1 was poorer than those with high expres-sion.The low expression of SHTN1 may be related to the poor prognosis of glioma patients,and the expres-sion of SHTN1 can be regulated by DNMT1 through DNA methylation.
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