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目的观察氨氯地平对小鼠黑色素瘤B16基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)和MMPs组织抑制因子(Tissue inhibitor of metalloproteinase-1,TIMP-1)表达的调节作用,并探讨其抑制肿瘤转移的作用机制。方法采用MTT法检测氨氯地平对B16细胞增殖的影响;建立C57BL/6小鼠黑色素瘤B16的自发性肺转移模型,随机分为阴性对照组(生理盐水)、阳性对照组(环磷酰胺20 mg/kg)、氨氯地平高[10 mg/(kg.d)]、中[3 mg/(kg.d)]、低[1 mg/(kg.d)]剂量组,采用明胶酶谱法检测肿瘤组织中MMP-9的活性;RT-PCR法检测肿瘤组织中MMP-9和TIMP-1基因mRNA的表达;免疫组化SP法检测肿瘤组织中MMP-9和TIMP-1蛋白的表达。结果氨氯地平呈时间和剂量依赖性抑制小鼠黑色素瘤B16细胞的生长,作用48 h的IC50为11.56μmol/L;氨氯地平可下调肿瘤组织中MMP-9的活性及MMP-9基因mRNA和蛋白的表达,上调TIMP-1基因mR-NA和蛋白的表达。结论氨氯地平通过调节MMP-9和TIMP-1基因的表达发挥其抑制肿瘤转移的作用。
Objective To investigate the regulatory effect of amlodipine on the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase-1 (TIMP-1) in mouse melanoma B16 Its mechanism of inhibiting tumor metastasis. Methods MTT was used to detect the effect of amlodipine on the proliferation of B16 cells. A spontaneous lung metastasis model of B16 melanoma in C57BL / 6 mice was established and randomly divided into negative control group (normal saline), positive control group (cyclophosphamide 20 (3 mg / (kg · d)], low [1 mg / (kg · d)], amlodipine [10 mg / (kg · d) The expression of MMP-9 and TIMP-1 mRNA in tumor tissue was detected by RT-PCR and the expression of MMP-9 and TIMP-1 in tumor tissue by immunohistochemistry . Results Amlodipine inhibited the growth of mouse melanoma B16 cells in a dose-dependent and time-dependent manner. The IC50 was 11.56 μmol / L for 48 h. Amlodipine could down-regulate the activity of MMP-9 and the expression of MMP-9 mRNA And protein expression, up-regulate TIMP-1 gene mR-NA and protein expression. Conclusion Amlodipine exerts its effect of inhibiting tumor metastasis by regulating the expression of MMP-9 and TIMP-1.