siRNA干扰LOXL2基因对人胃癌细胞株BGC823侵袭的影响及机制

目的 研究小干扰RNA(siRNA)抑制胃癌细胞中LOXL2的表达情况,并探讨LOXL2基因对人胃癌细胞株BGC823侵袭能力影响及机制.方法 QRTPCR及蛋白印迹(Western blot)技术检测人胃癌细胞株BGC823、正常胃上皮细胞株GES-1中LOXL2表达;合成针对LOXL2的siRNA,并转染胃癌细胞株BGC823;应用Transwell实验检测BGC823细胞侵袭能力;检测转染前后BGC823侵袭相关基因基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达变化.结果 LOXL2在BGC823细胞中的表达明显高于胃上皮细胞株(P<0?.01);与未转染和转染阴性对照质粒组相比转染组胃癌细胞BGC823中LOXL2 mRNA和蛋白的表达水平均明显降低(P<0.05),侵袭实验显示LOXL2-siRNA转染组胃癌细胞BGC823穿膜细胞数显著低于阴性对照质粒组和未转染组(P<0.05);转染后BGC823中MMP-2、MMP-9表达均较转染前降低(均P<0.05).结论 抑制胃癌细胞BGC823 LOXL2表达可降低细胞的侵袭能力.

Effect of RNA interference targeting at LOXL2 gene on the invasion of gastric cancer cell line-BGC823

Objective To observe the inhibition effects of small interference RNA ( siRNA) on the expression of LOXL2 in gastric cancer cells ,and to explore the effects and action mechanism of LOXL 2 gene on the invasion ability of human gastric cancer cell line-BGC823 .Methods The expression levels of LOXL 2 in gastric cancer tissues , adjacent tissues and human gastric cancer cell line-BGC823, gastric epithelial cell line-GES-1 were detected by Western Blot .Moreover LOXL2-siRNA was synthesized and tansfected into BGC 823 cells.The invasion ability of BGC823 cells was detected by Transwells assay.Besides the expression levels of MMP-2,MMP-9 were detected by qPCR and Western Blot before and after transfection.Results The expression levels of LOXL2 in BGC823 cells were significantly higher than those in GES-1 cells (P<0.01).The expression levels of LOXL2 mRNA and protein in BGC823 cells in LOXL2-siRNA group were significantly decreased,as compared with those in non -transfection group and negative plasmid control group ( P <0.05).The invasion experiment showed that the cell counts of breaking cell membrane in BGC 823 cells of LOXL2-siRNA group were significantly less than those in non -transfection group and negative plasmid control group ( P <0.05) .Moreover the expression levels of MMP-2,MMP-9 in BGC823 cells after transfection were significantly decreased ,as compared with those before transfection (P<0.05).Conclusion To inhibit the expression of LOXL2 in BGC823 cells can obviously decrease the invasion ability of BGC823 cells.