Effect of siRNA targeting protein arginine methyl transferase 5 on the proliferation and colony-formation in human gastric cancer SGC7901 cells

WU Xiaobin; TANG Futing; XU Hui

doi:10.3969/j.issn.1009-0460.2016.12.003

Abstract：Objective To explore the effect of small interfering RNA(siRNA) targeting protein arginine methyltransferase 5 (PRMT5) on the proliferation and colony-formation in human gastric cancer SGC7901 cells.Methods siRNA-1 and siRNA-2 designed for specific targeting PRMT5 were transiently transfected into SGC7901 cells.Meanwhile,antisense sequence was used as negative control(NC).48 h after transfection,Western blotting method was used to confirm the interfering efficacy to ensure that siRNA can be used for following experiments.CCK-8 assay was used to detect the proliferation of SGC7901 cells after gene-specific siRNA was transfected.EdU assay was used to examine the DNA replication of SGC7901 cells and plate colony-formation assay was used to evaluate the colony-formation of SGC7901 cells when PRMT5 was silenced by siRNA.Results The level of PRMT5 protein was significantly decreased in SGC7901 cells when siRNA-1 and siRNA-2 were transfected,suggesting that siRNA-1 and siRNA-2 could be used for further studies.The proliferation rate of SGC7901 cells was significantly reduced when PRMT5 was silenced(P＜0.01) in comparison with NC.Compared with NC,the positive rates of EdU cells interfered by siRNA-1 and siRNA-2 in PRMT5 group were(16.0±2.0)％ and (19.5 ± 3.0) ％,much lower than (38.0 ± 4.0) ％ in NC with significance (P ＜ 0.01).Furthermore,the numbers of SGC7901 colonies in PRMT5 group interfered by siRNA-1 and siRNA-2 were 50±6 and 68-±7,lower than 120± 11 of NC with significance(P＜ 0.01).Conclusion Knockdown of PRMT5 by gene-specific siRNA can inhibit the proliferation and colony-formation abilities in human gastric cancer SGC7901 cells,which provide new methods and theoretical basis for the treatment of gastric cancer.