基质金属蛋白酶（MMP）抑制剂LY52对人卵巢癌细胞系SKOV3中MMP-2和MMP-9表达及侵袭能力的抑制作用

[Inhibitory effects of matrix metalloproteinase (MMP) inhibitor LY52 on expression of MMP-2 and MMP-9 and invasive ability of human ovarian carcinoma cell line SKOV3].

作者信息

Yuan Yun-Xia, Xu Wen-Fang, Liu Jian, Chen Ming-Hui, Meng Hong, Qu Xian-Jun

机构信息

Department of Pharmacology, Faculty of Pharmacy, Shandong University, Jinan, Shandong, 250012, P. R. China.

出版信息

Ai Zheng. 2006 Jun;25(6):663-70.

PMID:16764758

Abstract

BACKGROUND & OBJECTIVE: Enhanced expression and activation of matrix metalloproteinase-2 (MMP-2) and MMP-9 are associated with tumor progression, invasion, and metastasis. Using synthetic MMP inhibitors to block the proteolytic activity of these enzymes is a potential strategy of treating cancer. This study was to observe the inhibitory effects of LY52, a synthetic specific MMP inhibitor, on the expression of MMP-2 and MMP-9 and invasive ability of human ovarian carcinoma cell line SKOV3.

METHODS

The inhibitory effect of LY52 on growth of SKOV3 cells was measured by MTT and clonogenic assays. The inhibitory effect of LY52 on the activity of gelatin was evaluated by spectrophotometry using succinylated gelatin as substrate. The inhibitory effect of LY52 on the expression of MMP-2 and MMP-9 was analyzed using SDS-PAGE zymography. The adhesive ability of SKOV3 cells to fibronectin (FN), laminin (LN), and matrigel was measured using MTT assay. The invasive ability of SKOV3 cells was assessed with a transwell cell culture chamber.

RESULTS

LY52 had weak cytostatic effect on SKOV3 cells, the inhibitory rates were 10.54%-37.66% within 120 h incubation when the concentration was 10 microg/ml. LY52 inhibited gelatin degradation via suppressing the activities of gelatinases, the 50% inhibitory concentration (IC50) was 11.9 microg/ml. When treated with 0.1, 1, 10, 100, or 1,000 microg/ml LY52 for 24 h, the expression of MMP-2 in SKOV3 cells was suppressed by 10.66%-31.47%, and the expression of MMP-9 was suppressed by 22.56%-56.71%. When treated with LY52 for 1 h, the maximum inhibitory rates of adhesion of SKOV3 cells to FN, LN, and matrigel were 29.79%, 48.94%, and 50.92%, respectively. When treated with 0.1, 1, 10, 100, and 1,000 microg/ml LY52 for 24 h, the inhibitory rates of invasion of SKOV3 cells were 7.5%, 42.07%, 66.54%, 72.15%, and 82.84%, respectively.

CONCLUSION

LY52 suppresses the invasion of SKOV3 cells via inhibiting the expression of MMP-2 and MMP-9.

摘要

背景与目的

基质金属蛋白酶-2（MMP-2）和MMP-9的表达增强及激活与肿瘤进展、侵袭和转移相关。使用合成的MMP抑制剂阻断这些酶的蛋白水解活性是一种潜在的癌症治疗策略。本研究旨在观察合成的特异性MMP抑制剂LY52对人卵巢癌细胞系SKOV3中MMP-2和MMP-9表达及侵袭能力的抑制作用。

方法

采用MTT法和克隆形成试验检测LY52对SKOV3细胞生长的抑制作用。以琥珀酰化明胶为底物，用分光光度法评估LY52对明胶活性的抑制作用。采用SDS-PAGE酶谱分析法分析LY52对MMP-2和MMP-9表达的抑制作用。用MTT法检测SKOV3细胞与纤连蛋白（FN）、层粘连蛋白（LN）和基质胶的黏附能力。用Transwell细胞培养小室评估SKOV3细胞的侵袭能力。

结果

LY52对SKOV3细胞有较弱的细胞生长抑制作用，浓度为10μg/ml孵育120 h内抑制率为10.54% - 37.66%。LY52通过抑制明胶酶活性抑制明胶降解，50%抑制浓度（IC50）为11.9μg/ml。用0.1、1、10、100或1000μg/ml LY52处理24 h后，SKOV3细胞中MMP-2的表达被抑制10.66% - 31.47%，MMP-9的表达被抑制22.56% - 56.71%。用LY52处理1 h，SKOV3细胞对FN、LN和基质胶黏附的最大抑制率分别为29.79%、48.94%和50.92%。用0.1、1、10、100和1000μg/ml LY52处理24 h后，SKOV3细胞侵袭的抑制率分别为7.5%、42.07%、66.54%、72.15%和82.84%。