KISS-1及MMP-9基因在胃癌侵袭转移中的作用及相关性

Effect and relationship of KISS-1 and Matrixmetalloproteinase-9 on invasion and metastasis of gastric carcinoma

目的探讨肿瘤转移抑制基因KISS-1在胃癌侵袭、转移中的作用及其与MMP-9表达的关系。方法采用Lipofectamine脂质体介导将pcDNA3.1-KISS-1真核表达质粒转染胃癌BGC-823细胞,经G418筛选,建立稳定高表达KISS-1蛋白的细胞系,通过RT-PCR和Western blotting法证实转染成功,并检测细胞中MMP-9蛋白及mRNA的表达情况;采用Transwell体外侵袭实验,探讨KISS-1对胃癌细胞株侵袭能力的影响。结果 Western blotting和RT-PCR结果显示:转基因组BGC-823细胞中KISS-1蛋白及mRNA的表达与转空质粒组和对照组相比均明显增加(P均<0.05);转基因组BGC-823细胞中MMP-9蛋白及mR-NA的表达与转空质粒组和对照组相比均明显降低(P均<0.05);Transwell检测结果显示:转基因组BGC-823细胞的穿膜数与转空质粒和对照组相比均明显降低(P<0.05),侵袭力抑制率达到25%。结论 pcDNA3.1-KISS-1的有效转染可降低MMP-9的表达并对胃癌细胞BGC-823的侵袭能力具有抑制作用。KISS-1基因对胃癌侵袭转移的抑制作用可能是通过下调MMP-9的功能实现的。

Purpose To investigate the effect and relationship of metastasis suppressor gene KISS-1 and matrixmetalloproteinase-9 （MMP-9） on invasion and metastasis of gastric carcinoma. Methods The eukaryotic expression vector pcDNA3.1-KISS-1 was trans- fected into gastric carcinoma cell line BGC-823 by lipofectamine. To observe the expression of MMP-9 （ Western blotting and RT-PCR） and potential of invasion （Transwell in vitro） in transfected BGC-823. Results After pcDNA3.1-KISS-1 recombinant vector was suc- cessfully constructed in BGC-823 cells, the expressions of KISS-1 protein and mRNA in cells transfected with pcDNA3. 1-KISS-1 were significantly higher than it in cells transfected with pcDNA3.1 and control cells （ P 〈 0. 05 ）. The expression of MMP-9 protein and mRNA in cells transfected with pcDNA3.1-KISS-1 were significantly lower than it in cells transfected with pcDNA3. 1 and control cells （ P 〈 0. 05 ）. Cell number invading through Matrigel filter in cells transfected with pcDNA3.1-KISS-1 was significantly decreased com- pared with that in cells transfected with pcDNA3. 1 and control group （ P 〈 0. 05 ）. The inhibition rate of invading ability reached 25%. Conclusion The effective transfection of pcDNA3, l-KISS-1 can decrease the expression of MMP-9 and inhibit the potential of invasion of BGC-823 cells. The depressant effect of KISS-1 gene on invasion and metastasis of gastric carcinoma may implement by de- creasing the action of MMP-9.