细丝蛋白A抑制人结肠癌SW480细胞体外侵袭转移能力

Inhibitory effect of filamin A on invasion and metastasis of human colon carcinoma cell line SW480 in vitro

目的研究细丝蛋白A(FLNa)对人结肠癌SW480细胞侵袭转移行为的影响,并初步探讨其抑癌的机制。方法将FLNa基因质粒载体pcDNA3.1/V5-His-TOPO/FLNa,以脂质体介导方式转染SW480细胞。经G418筛选,获得FLNa基因稳定表达的SW480/FLNa细胞。反转录聚合酶链反应(RT-PCR)和Western blot检测FLNa基因导入;RT-PCR和Western blot检测细胞中基质金属蛋白酶-9(MMP-9)的表达;MTT法检测细胞的增殖能力;划痕损伤实验、Transwell小室和Matrigel侵袭模型评价FLNa对细胞侵袭和转移能力。结果在SW480细胞中,FLNa的表达载体pcDNA3.1/V5-His-TOPO/FLNa获得稳定表达,SW480/FLNa细胞FLNa的mRNA和蛋白的表达水平均较SW480细胞高,分别为[(1.27±0.03)vs(0.14±0.02),P<0.01]和[(1.18±0.03)vs(0.25±0.02),P<0.05];SW480/FLNa细胞MMP-9的mRNA和蛋白表达水平均低于SW480细胞,分别为[(0.19±0.02)vs(1.63±0.04),P<0.05]和[(0.12±0.02)vs(0.79±0.04),P<0.05]。结论 FLNa抑制SW480细胞的侵袭和转移能力,其机制可能与降低MMP-9的表达有关。

Objective To explore the inhibitory effect of Filamin A（FLNa） on invasion and metastasis of human colon carcinoma cell line SW480 and investigate its mechanisms.Methods Expressing vector pcDNA3.1/V5-His-TOPO/ FLNa was transfected into SW480 cells by lipofectamine protocols.After G418 selection,the SW480/FLNa cells expressing FLNa stably were obtained.Expression of FLNa in tumor cells was confirmed by RT-PCR and Western blot.RT-PCR assay and Western blot were also employed to detect the expression of matrix metalloproteinases-9（MMP-9）.Cell proliferation in vitro was assessed by MTT assay.Inhibitory effect of filamin A on SW480 cells wasobserved by Wound healing assay,Transwell chamber experiment,and Matrigel invasion assay.Results The results of RT-PCR and Western blot demonstrated that the recombinant vector was transfected into SW480 cells by liposome.The expressions of MMP-9 mRNA and protein were lower in the SW480/FLNa cells than that in the wild SW480 cells;The inhibitory effect of FLNa on invasion and metastasis of SW480 cells was proved by wound healing assay,Transwell chamber experiment and matrigel invasion assay.Conclusion FLNa has significant inhibitory effect on the invasion and metastasis of human colon carcinoma cell line SW480.The mechanism might be explained by suppressive effect of MMP-9.