Modomics - A Database of RNA Modifications

Published on Dec. 1, 2017 in Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi volume 33(12).

PubMed ID: 29382422


Objective To investigate the effect of knockdown of YTH N(6)-methyladenosine RNAbinding protein 2 (YTHDF2) on cell proliferation, cell cycle and apoptosis ofMGC-803 human gastric cancer cells in vitro. Methods The TCGA database wasdownloaded from UCSC Cancer Browser and to search for the differential expressionsof YTHDF2 mRNA in gastric cancer tissues. Short hairpin RNA (shRNA) targeting YTHDF2was designed and cloned into lentivirus expression vector pLKO.1. Furthermore,MGC-803 gastric cancer cells were transfected with pLKO.1-shRNA to knockdown theexpression of YTHDF2, which was confirmed by the detection of YTHDF2 mRNA andprotein expression using real-time quantitative PCR and Western blotting,respectively. Then cell proliferation was observed by CCK-8 assay, and cell cycleand apoptosis were examined by flow cytometry. Results According to the TCGAdatabase, the expression of YTHDF2 mRNA in gastric cancer was significantly higherthan that in the normal tissues. MGC-803 stably expressing YTHDF2-shRNA wassuccessfully established. Furthermore, the proliferation capacity ofYTHDF2-shRNA-expressing MGC-803 cells was significantly inhibited compared with thecontrols. Similarly, the percentage of YTHDF2-shRNA-expressing MGC-803 cells in G1phase increased and in S phase decreased compared with the controls. Meanwhile,apoptosis ratio of YTHDF2-shRNA-expressing MGC-803 cells was significantly highercompared with the control groups. Conclusion Knockdown of YTHDF2 in MGC-803 cellsinhibits cell proliferation and promotes apoptosis.