LAP3调控MMP-2/9的表达对人皮肤恶性黑色素瘤细胞侵袭转移的影响

The Inhibition of Leucine Aminopeptidase 3 Suppresses Invasion of HumanMelanoma Cells Through Down-Regulation of MMP-2/9

目的探讨亮氨酸氨肽酶3(LAP3)基因沉默对人皮肤恶性黑色素瘤细胞增殖、侵袭及其迁移的影响。方法在黑色素瘤细胞中转染LAP3 siRNA,实验分为空白对照组、阴性对照组、LAP3干扰组,采用克隆形成实验、Transwell小室和划痕实验分别检测黑色素瘤细胞的增殖、侵袭和迁移能力,免疫印迹法检测黑色素瘤细胞中LAP3、MMP-2/9、上皮型钙黏蛋白、N-钙黏附素、波形蛋白的表达。结果在LAP3干扰后,黑色素瘤细胞A2058的存活率、克隆形成率、迁移细胞数、侵袭细胞数以及LAP3、MMP-2/9、N-钙黏附素、波形蛋白的表达水平均显著降低(P<0.05),而细胞中上皮型钙黏蛋白的表达水平显著升高(P<0.05)。结论LAP3基因沉默可抑制黑色素瘤细胞的增殖,并通过减弱下游蛋白MMP-2/9的表达以及细胞上皮间质转化进程协同抑制黑色素瘤细胞侵袭和迁移。

Objective To investigate the effects of leucine aminopeptidase 3(LAP3)gene silencing on proliferation,invasion and migration of melanoma cells.Methods LAP3 siRNA was transfected into melanoma cells,and the experiment was divided into the normal control group(NC),Si-normal control group(SiNC)and SiLAP3 group(SiLAP3).Clone formation assays,transwell migration assays and scratch healing assay were conducted to detect the proliferation,invasion and migration ability of melanoma cells,respectively.Western blotting analysis was used to detect the expression levels of LAP3,MMP-2/9,E-cadherin,N-cadherin and Vimentin proteins in melanoma cells.Results Compared with the normal control group and Si-negative control group,cell survival rate,clone formation rate,number of invasion cells,number of migration cells,and the expression levels of LAP3,MMP-2/9,N-cadherin and vimentin protein of the SiLAP3 group were all significantly reduced(all P<0.05),while the expression levels of E-cadherin protein in cells were significantly higher than other groups.(P<0.05).Conclusion Silencing of LAP3 gene inhibits the proliferation of melanoma cells,and also inhibits invasion and migration of melanoma cell via attenuating the expressing levels of MMP-2/9 and the process of cell epithelial-mesenchymal transition.