siRNA干扰LOXL2基因对人胃癌细胞株BGC823侵袭的影响及机制

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

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。

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.