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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 study the effect of small interfering RNA (siRNA) on the expression of LOXL2 in gastric cancer cells and to explore the effect of LOXL2 gene on the invasiveness of human gastric cancer cell line BGC823. Methods QRTPCR and Western blot were used to detect the expression of LOXL2 in human gastric cancer cell line BGC823 and normal gastric epithelial cell line GES-1. SiRNA targeting LOXL2 was transfected into BGC823 gastric cancer cell line. The invasion of BGC823 cells was detected by Transwell assay The expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in BGC823 cells was detected before and after transfection. Results The expression of LOXL2 in BGC823 cells was significantly higher than that in gastric epithelial cells (P <0.01). Compared with untransfected and transfected negative control plasmids, LOXL2 mRNA and protein expression levels were significantly increased in BGC823 cells (P <0.05). The invasion assay showed that the number of transmembrane cells in gastric cancer cells BGC823 in LOXL2-siRNA transfected group was significantly lower than that in negative control plasmid group and untransfected group (P <0.05) MMP-9 expression were lower than before transfection (all P <0.05). Conclusion Inhibition of LOXL2 expression in BGC823 gastric cancer cells can reduce the invasiveness of cells.