粘蛋白MUC2反义核酸抑制胃癌细胞的研究

Inhibitory effects on human gastric cancer cell line by MUC2 antisense oligodeoxynucleotide

目的 :探讨粘蛋白MUC2反义脱氧寡核苷酸 (ASODN)对人胃癌细胞株SGC790 1的抑制作用。方法 :采用人工合成的MUC2ASODN经阳离子脂质体包裹后转染入胃癌株SGC790 1细胞 ,采用MTT法、形态学观察及免疫组化法测定MUC2ASODN对胃癌细胞的抑制作用。结果 :MTT法测细胞增殖活性见不同浓度ASODN均能抑制SGC790 1细胞的增殖 ,抑制作用在 4 8h最强 ,0 .5 μmol/LASODN对细胞的抑制率为 5 5 % ,时间延长抑制作用逐渐减弱。SGC790 1细胞转染MUC2ASODN后 ,与对照组相比 ,光镜下观察到细胞数量减少、体积变小、核分裂明显减少、可见较多的变性、坏死。免疫组化S -P法染色提示转染ASODN后 ,SGC790 1细胞mRNA的翻译受到抑制 ,MUC2蛋白表达水平明显降低 ,p16蛋白表达明显增强 ,粘附分子CD4 4V6表达明显降低 ,MMP - 2的表达也显著下降 ,MMP - 9的表达无明显变化。结论 :使用人工合成MUC2ASODN能有效抑制胃癌细胞株SGC790 1的增殖及侵袭。

Objective:To investigate the inhibitory effect in vitro of mucin gene MUC2 antisense oligodeoxynucleotide (ASODN) on its gene expression and cell proliferation on gastric cancer cell line.Methods:Phosphorothioate MUC2 ASODN were synthesized and transfected to SGC7901 cells mediated by lipofectin.Their inhibitory effects on cell proliferation were determined by MTT method,light microscopy and immunohistochemical method.Results:The determination by MTT method demonstrated that MUC2 ASODN of varied concentrations could significantly inhibit the growth of SGC7901 cell lines while the control lipofectin showed no such effect.The result also suggested that the inhibitory effect was dose-dependent and time-dependent.The inhibition peaked at 48th hour after transfection,and the inhibition rate reached 55% when the concentration was 0.5μmol/L.After transfecting with MUC2 ASODN,SGC7901 cells showed decrease in number,volume and karyokinesis,increase in necroses under light microscopy.And the test by immunohistochemical method indicated that ASODN could downregulate the expression levels of MUC2 protein,MMP-2 protein and CD44V6 protein,but upregulate the expression levels of p16 protein.Conclusion:The data suggest that MUC2 ASODN transfection could effectively inhibit SGC7901 cells proliferation and invasion and metastasis potential of SGC7901 cells.