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目的研究钙调素拮抗剂EBB抑制HT1080人纤维肉瘤细胞侵袭的作用。方法采用四唑盐(MTT)比色试验测定药物敏感性,Zymogrophy测定基质金属蛋白酶-2(MMP-2)及MMP-9明胶酶活性,逆转录-聚合酶链反应(RT-PCR)检测MMP-2、MMP-9及基质金属蛋白酶抑制因子-1(TIMP-1)mRNA表达,Transwell小室进行细胞侵袭分析。结果MTT测得半数抑制浓度(IC50)为(8.2±1.2)μg/ml。钙调素拮抗剂EBB使HT1080细胞的侵袭能力明显下降(P<0.001),表现为MMP-2和MMP-9基因表达及酶活性下降。同时TIMP-1的基因表达量相应升高。结论钙调素拮抗剂EBB可以降低HT1080人纤维肉瘤细胞的侵袭能力,基质金属蛋白酶分泌受抑制可能是抑制细胞转移的机制之一。
Objective To investigate the role of calmodulin antagonist EBB in inhibiting the invasion of HT1080 human fibrosarcoma cells. Methods MTT colorimetric assay was used to determine the drug sensitivity. Zymogrophy was used to determine the activity of MMP-2 and MMP-9 gelatinase. MMP-9 was detected by reverse transcription-polymerase chain reaction (RT-PCR) -2, MMP-9 and TIMP-1 mRNA expression were detected by Transwell assay. Results The median inhibitory concentration (IC50) measured by MTT was (8.2 ± 1.2) μg / ml. The calmodulin antagonist EBB significantly decreased the invasiveness of HT1080 cells (P <0.001), indicating that the expression of MMP-2 and MMP-9 decreased and the activity of the enzyme decreased. At the same time, the gene expression of TIMP-1 increased accordingly. Conclusions Calmodulin antagonist EBB can reduce the invasion ability of HT1080 human fibrosarcoma cells, and inhibition of matrix metalloproteinase secretion may be one of the mechanisms of inhibiting cell metastasis.