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Expression of DNMT1 in gastric carcinoma tissue and its effects on proliferation and migration of gastric carcinoma cells

ZHOU Yi, QIN Jian, LI Xu, ZHU Lin, LI Ji-kun   

  1. Department of General Surgery， Shanghai General Hospital， Shanghai Jiao Tong University School of Medicine， Shanghai 200080， China
  • Online:2016-07-28 Published:2016-08-31
  • Supported by:

    National Natura Science Foundation of China， 81472236； Foundation of Science and Technology Commission of Shanghai Municipality， 14ZR1433400

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Abstract:

Objective To explore the effects of DNA methyl transferase 1 (DNMT1) on the proliferation and migration of gastric carcinoma cells and its possible mechanisms. Methods The mRNA levels of DNMT1 in cancer tissues and adjacent normal tissues of 60 patients with gastric carcinoma were detected with qRT-PCR. Lentivirus packing was used to establish MKN45, SGC7901, and MKN28 gastric carcinoma cell lines expressing stable DNMT1-shRNA, which served as the transfection group. The untreated MKN45, SGC7901, and MKN28 cell lines served as the untransfected group and the MKN45, SGC7901, and MKN28 cell lines treated with lentivirus control served as the negative control group. Proliferation of MKN45, SGC7901, and MKN28 cells was detected by CCK8 method. The expression of CDC25B was measured by Western blotting. The effect of DNMT1 on the migration of gastric carcinoma cells was detected by cell migration assay. Results The mRNA levels of DNMT1 in 52 patients (86.7%，52/60) were up-regulated in cancer tissues than in adjacent normal tissues and the mRNA levels of DNMT1 in 17 of them (28.3%, 17/60) were up-regulated more than two times. The transcription level of DNMT1 in cancer tissues of patients with lymph node metastasis, vascular invasion, and TNM stage Ⅲ-Ⅳ was up-regulated significantly (P＜0.05). The growth of MKN45, SGC7901, and MKN28 cells was significantly inhibited 3 d after DNMT1-shRNA transfection. The absorbance values of three cell lines in the transfection group were significantly lower than those in the untransfected group and the negative control group (P=0.000). The protein expression of CDC25B and the migration ability decreased in the transfection group as compared with the untransfected group and the negative control group (both P＜0.05). Conclusion The expression of DNMT1 is significantly higher in gastric carcinoma tissues than that in adjacent normal tissues. DNMT1 can promote the proliferation and metastasis of gastric carcinoma cells, which may be associated with the regulation on CDC25B.

Key words: stomach neoplasms, cell proliferation, cell migration, DNMT1, CDC25B