The Study Of The Effect Of Allicin On Growth Of Mouse Ascites Melanoma Cell B16-F1 And Of The Proliferative Role Of Mouse Melanoma Cell On The Lymphangiogenesis In Vitro

Objective To establish an animal model of benign lymphangiomas in mice. observe the impact of mouse ascites melanoma cell B16-F1 on the lymphangongenesis. To study the effect of allicin on the lymphangongenesis and the growth and apoptosis of mouse ascites melanoma cell line B16-F1.Methods Sixteen C57BL/6 mice accepted twice intraperitoneal injection with incomplete Freund's adjuvant with 15 days interval. Neoplasms induced were studied histopathologically. The expressions of VEGF-C and Flt-4 were examined by immunohistochemical method in neoplasms specimen. The neoplasm was harvested and embedded in fibrin gel to culture with conditioned medium of B16-F1 cell and different concentration of allicin in vitro. After exposed to different concentration of allicin for different time, the survival rate of B16-F1 cells was explored by MTT. Terminal deoxylnucleotide transferase directed X-UTP nick and end labelling (TUNEL) was used to observe apoptosis. RT-PCR was used to study the expression of ras gene.Results White solid tumor masses were induced in peritoneal cavity in 13 of 16 mice (81.3%). The histological study of tumors was lymphangioma, and tumor cells expressed VEGF-C and Flt-4 positively. Lymphatic capillary generating from lymphangioma specimen in the gel was observed and the conditioned medium of B16-F1 cell was observed to enhance the growth of the vessels, but the growth of the vessels could be inhibited by allicin. Compared with the experimental control group, allicin showed significant inhibiting effect on B16-F1 cells at the dosage of 3-15 μ g/ml. When the dosage of allicin was low, the apoptotic rate of cells was significantly increased,the expression of ras gene was significantly decreased.Conclusion Intraperitoneal injection with incomplete Freund's adjuvant is a good method to induce benign lymphangiomas in mice. B16-F1 cell can accelerate lymphangiogenesis, which warrants further study of the role of lymphangiogenesis in metastasis of melanoma. Allicin can inhibit the growth of lymphangiogenesis and mouse ascites melanoma cell line B16-F1, allicin can also induce the apoptosis. The expression of ras gene may be decreased by allicin.