NS398对SGC7901胃癌细胞生长及CD44V6、MMP-9基因表达的影响

The Effect of NS398 on the Growth of SGC7901 Gastric Cell Line and the Expression of CD44V6、MMP-9 Gene

目的探讨COX-2特异性抑制剂NS398对SGC7901胃癌细胞体外生长及CD44V6、MMP-9基因表达的影响。方法体外培养SGC7901胃癌细胞,应用免疫细胞化学方法,检测SGC7901胃癌细胞中COX-2、CD44V6、MMP-9蛋白表达水平,酶联免疫吸附试验(ELISA)检测NS398对胃癌细胞PGE2分泌水平的影响,MTT法检测NS398对SGC7901胃癌细胞体外生长的抑制效应,半定量RT-PCR检测NS398作用后胃癌细胞株中CD44V6、MMP-9基因表达水平的变化。结果 SGC7901胃癌细胞中COX-2、CD44V6、MMP-9呈阳性表达,NS398可显著抑制SGC7901胃癌细胞PGE2的分泌,200μmol/L NS398抑制率达64.3%,NS398对胃癌细胞生长抑制作用呈时间及浓度依赖性;NS398作用24 h后胃癌细胞中CD44V6、MMP-9基因表达水平下降,200μmol/L NS398对CD44V6、MMP-9基因表达抑制率分别为45.7%、43.6%。结论 SGC7901胃癌细胞中COX-2、CD44V6、MMP-9呈阳性表达,NS398可有效抑制SGC7901胃癌细胞PGE2的分泌和胃癌细胞生长,并可抑制与胃癌转移相关的CD44V6、MMP-9基因的表达。

Objective To study the expression of CD44V6 and MMP-9 in human gastric cancer,and the effects of NS-398,a specfic cyclooxygenase-2（COX-2） enzyme inhibitor on the cell growth and the mRNA expression of CD44V6 and MMP-9 gene in gastric cancer cell line SGC7901.Methods The expression of COX-2,CD44V6 and MMP-9 protein in the SGC7901 gstric cancer cells were detected by immunohistochemical technique;The effect of NS398 on the inhibition of PGE2 serection in SGC-7901 gastric cancer cells were detected by ELISA;the inhibition effect of NS398 on growth of SGC7901 gastric cell line were measured by MTT;The expression of CD44V6 and MMP-9 mRNA in the SGC7901 gastric cancer cells before and after NS-398 treatment were detected by semi-quantitative RT-PCR.Results Protein expression of COX-2、CD44V6 and MMP-9 were positive in SGC7901 gastric cancer cells.The PGE2 production was significantly inhibited by NS-398,while a 64.3% inhibition rate was achieved by 200 μmol/L NS-398 treament.The inhibition on the PEG2 serection was time and concentration dependent.24 h after NS-398 treatment,the mRNA expression of CD44V6 and MMP-9 downregulated.When the concentration of NS-398 is 200 mmol/L,the expression of CD44V6 and MMP-9 mRNA were inhibited to 45.7% and 43.6% respectively.Conclusion Protein expression of COX-2 、CD44V6 and MMP-9 are positive in SGC7901 gastric cancer cells.NS-398 can inhibit not only the cell growth and PGE2 serecttion,but also the mRNA expression of CD44V6 and MMP-9 gene.