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Archive > Volume 25 Issue 11 > 2018,25(11):1119-1124. DOI:10.3872/j.issn.1007-385X.2018.11.006 Prev Next
Basic Research
Effects of nuclear factor 5 of activated T cells on proliferation and apoptosis of human gastric cancer MGC803 cells
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Abstract:
Objective: To investigate the effects of nuclear factor 5 of activated T cells (NFAT5) on proliferation and apoptosis of human gastric cancer MGC803 cells and to explore the possible mechanisms. Methods: Three siRNAs targeting NFAT5 gene (siRNA2567,siRNA2714 and siRNA4562) and one negative control siRNA were designed and chemically synthesized before transfected into human gastric cancer cell line MGC803 by liposome. Real-time PCR was used to detect the changes of NFAT5 mRNA expression in MGC803 cells to further pick out the siRNA that most effectively inhibit the expression of NFAT5. Further, Real-time PCR and Western blotting assay were carried out to test mRNA and protein levels of NFAT5 and S100A4 in cells 48 h after NFAT5-siRNA transfection.Then, CCK-8 assay and FCM assay were used to detect the influence of silencing NFAT5 on cell proliferation and apoptosis, respectively.Results: siRNA2567 was the most effective siRNA that significantly inhibited the expression of NFAT5 mRNA (P<0.01),and thus was validated as NFAT5-siRNA. Real-time PCR and Western blotting assay confirmed that both mRNA and protein levels of NFAT5 and S100A4 were down-regulated in cells 48 h after NFAT5-siRNA transfection. Compared with NC-siRNA group, the proliferation ability of MGC803 cells in the NFAT5-siRNA group was significantly down-regulated at 72 h and 96 h (P<0.01). And FCM assay showed that compared with NC-siRNA group, cell apoptosis rate of NFAT5-siRNA group was significantly increased from (2.7±0.2)% to (7.9±0.2)%, (P<0.01) 48 h after NFAT5-siRNA transfection. Conclusion: NFAT5-siRNA transfection can silence NFAT5 gene expression in gastric cancer MGC803 cells effectively. NFAT5 may inhibit proliferation and promote cell apoptosis of gastric cancer cells possibly through regulating S100A4 expression.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81803855), and the Scientific Research Foundation of Education Commission of Liaoning Provincial （No.L201614）
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