Regulation Of Amlodipine On Expression Of MMP-9 And TIMP-1 In Murine Melanoma B16

Objective To observe the regulation of Amlodipine on expression of MMP-9,TIMP-1, and explore the anti-metastasis mechanism in murine melanoma B16 of the drug. Method To detect the growth inhibitory rate of Amlodipine on murine melanoma B16 cells by MTT assay and to calculate its IC50.B16 cells were allowed to grow in cell culture which concentration were 1/2 IC50,IC50,without Amlodipine were used as control,respectively. Supernatant were collected after 24h and MMP-9 activity was measured by Gelatin zymography. The spontaneous pulmonary metastasis models were established in C57BL/6J mice. The mice were equally divided into control group, CTX (20mg/kg .2d ip.)group, and three treatment groups of high(10 mg/kg.d po), middle(3 mg/ kg.d po) , and low (1 mg /kg.d po) dose of Amlodipine. To weight the tumor and take count of metastasis tubercular on the surface of lung,calculate the inhibitory rate and metastasis rate of amlodipine on the tumor. Gelatin zymography,immunohistochemistry and RT-PCR analysis were used to detect the expression of MMP-9,TIMP-1 in tumor tissue. Result (1)With dose and time dependently, Amlodipine inhibit the growth of murine melanoma B16 cells.(2) Gelatin zymography showed that the expression of MMP-9 in treatment group were lower than that in control group.The activity of MMP-9 in the supernatant was could not been detected.(3)The proliferation and the spontaneous pulmonary metastasis of murine melanocytoma B16 were inhibited by amlodipie, in a dose-dependent manner.(4)The result of immunohistochemistry confirmed that the expression of MMP-9 in control group were significantly more than that of treatment group.(5)Amlodipine inhibited the expression of mRNA of MMP-9,while increased the expression of mRNA of TIMP-1. Conclusions(1)Amlodipine suppresses the growth of melanoma B16 cells in vitro. (2)Amlodipine may suppress the tumor metastasis via regulating the expression of MMP-9,TIMP-1.