Study Of Amlodipine On Murine Melanoma B16 Metastasis

Objective:To observe the anti-metastasis of CCB-amlodipine on murine melanocytoma B16 and to explore the related mechanisms. And to provide the theoretical basis of calcium channel blocker (CCB) on tumor chemotherapy for clinical applications.Methods: 64 C57BL/6J mice(paroecious equally)are inoculated with murine melanocytoma B16 cell ( 2×106/mouse ) in the fold inguen subcutaneously,and on the second day of the inoculation,the mice were divided into 4 groups and the treatment was begin:①control group:the mice were treated with NS 0.2ml/mouse.②amlodipine low dose group:the mice were treated with amlodipine1.0mg/kg.③amlodipine middle dose group:the mice were treated with amlodipine 3.0mg/kg.④amlodipine high dose group:the mice were treated with amlodipine 10mg/kg respectively. The treatment was totally 21days. On the second day of the last treatment,put the mice to death by cut off the net, observe the pulmonary metastasis and the inhibitory rate of the tumor; In vitro study:Study of the inhibitiory effect of platelet aggregation. To set up the immune adherence mode of the platelet to murine melanocytoma B16 cells,and to compare the ability of the platelet adhere to murine melanocytoma B16 cells before and after the drugs administration. Immunohistochemical was introduced to detect the expression of IL-8 protein levels of the tumors in treatment group and control group; RT-PCR was used to detect the expression of IL-8mRNA in the tumor tissues; Western Blot was used to observe the expression of ERK1/2, phosphorylated ERK1 / 2 (P-ERK1 / 2) in tumor tissues.Results:In addition to inhibit the proliferation and the spontaneous pulmonary metastasis of murine melanocytoma B16 cells ,anlodipine inhibits the aggression of the platelets induced by the murine melanoma-B16 cells and platelet adhere to murine melanocytoma B16 cells,and in a dose-dependent manner;the expression of IL-8 protein , IL-8 mRNA and p-ERK1/2 was significantly reduced in the amlodipine treatment groups,in a dose dependent manner;while there was no significant variation in the expression of ERK1/2.Conclusion: Amlodipine exhibits the effects of anti-proliferatiion and anti-spontaneous pulmonary metastasis on murine melanocytoma B16 cells,the relative mechanism may be as follows: Firstly, the inhibition of the platelet aggression induced by the murine melanoma-B16 cells and platelet adhere to melanoma-B16 cells. Secondly, amlodipine affected the transcription and expression of IL-8 by inhibition of ERK1/2 signaling pathway .