敲低MGC-803胃癌细胞YTHDF2抑制其增殖并促进其凋亡

Knockdown of YTH N^6-methyladenosine RNA binding protein 2 ( YTHDF2 ) inhibits proliferation and promotes apoptosis in MGC-803 gastric cancer cells

目的研究敲低YTH结构域N^6甲基腺嘌呤(m^6A)RNA结合蛋白2(YTHDF2)对MGC-803人胃癌细胞的增殖、细胞周期和细胞凋亡的影响。方法用UCSC Cancer Browser下载癌症基因组图谱(TCGA)数据库筛查YTHDF2 mRNA在胃癌中的表达情况。设计并构建能够靶向敲低YTHDF2的短发夹RNA(shRNA)病毒载体;病毒感染MGC-803细胞,敲低YTHDF2水平后,实时荧光定量PCR检测YTHDF2 mRNA水平,Western blot法检测YTHDF2蛋白水平,CCK-8法检测细胞增殖,流式细胞术检测细胞周期和细胞凋亡。结果通过TCGA数据库查询发现YTHDF2在胃癌组织中高表达。成功构建YTHDF2敲低的稳定转染MGC-803细胞。与对照组相比,YTHDF2敲低后,明显抑制细胞增殖;G1期细胞明显增加,S期细胞明显减少;细胞凋亡率升高。结论敲低MGC-803胃癌细胞YTHDF2水平,抑制胃癌细胞增殖并促进其凋亡。

Objective To investigate the effect of knockdown of YTH N^6-methyladenosine RNA binding protein 2（ YTHDF2）on cell proliferation,cell cycle and apoptosis of MGC-803 human gastric cancer cells in vitro. Methods The TCGA database was downloaded from UCSC Cancer Browser and to search for the differential expressions of YTHDF2 mRNA in gastric cancer tissues. Short hairpin RNA（ shRNA） targeting YTHDF2 was designed and cloned into lentivirus expression vector p LKO. 1. Furthermore,MGC-803 gastric cancer cells were transfected with p LKO. 1-shRNA to knockdown the expression of YTHDF2,which was confirmed by the detection of YTHDF2 mRNA and protein expression using real-time quantitative PCR and Western blotting,respectively. Then cell proliferation was observed by CCK-8 assay,and cell cycle and apoptosis were examined by flow cytometry. Results According to the TCGA database,the expression of YTHDF2 mRNA in gastric cancer was significantly higher than that in the normal tissues. MGC-803 stably expressing YTHDF2-shRNA was successfully established. Furthermore,the proliferation capacity of YTHDF2-shRNA-expressing MGC-803 cells was significantly inhibited compared with the controls. Similarly,the percentage of YTHDF2-shRNA-expressing MGC-803 cells in G1 phase increased and in S phase decreased compared with the controls. Meanwhile,apoptosis ratio of YTHDF2-shRNA-expressing MGC-803 cells was significantly higher compared with the control groups. Conclusion Knockdown of YTHDF2 in MGC-803 cells inhibits cell proliferation and promotes apoptosis.