The Mechanism Of Platelet Membrane Glycoprotein GPIbα In Tumor Metastasis

Part ⅠConstruction of lung metastasis model of melanoma in C57BL/6J mice【Objective】To investigate the influencing factors involved in the establishment of melanoma lung metastasis model in C57BL/6J mice,including the way of tumor inoculation,the number of inoculated cells and the time of tumor formation.【Methods】Mouse melanoma cells B16F10 were cultured in vitro.1)Eighteen male mice were randomly divided into three groups.Mice in each group were received by 100 μL cell suspension(including 3 ×10~6 melanoma cells)intravenous,intraperitoneal and subcutaneous injection,respectively.After two weeks,mice were dissected and the tumor growth and metastasis were observed;2)Eighteen male mice were randomly divided into three groups.Mice in each group were injected with 3 ×10~6 cells,1 ×10~6 cells,and 3 ×10~5 cells through tail vein,respectively.After two weeks,mice were dissected and the tumor growth and metastasis were observed;3)Eighteen male mice were randomly divided into three groups.Mice in each group was injected with 1 ×10~6 cells though tail vein.Mice were dissected after one week,two weeks and three weeks,respectively.The growth and metastasis of tumor was observed.【Results】1)The success rate of lung metastasis was 100% in mice by intravenous injection,but the lung metastasis was not found in mice by intraperitoneal injection and subcutaneous injection.2)The size of metastatic melanoma nodules were moderate in mice inoculated by 1 ×10~6 cells;the number of melanoma cells with lung metastasis was too much in mice inoculated by 3 ×10~6 cells;the number of melanoma cells with lung metastasis was too small in mice inoculated by 3 ×10~5 cells.3)The significant metastasis of melanoma cells were observed in mice dissected after two weeks without mice death;the number of melanoma cells in lung was too much in mice dissected after three weeks,the number ofmelanoma cells in lung was too small in mice dissected after one week.【Conclusion】1 ×10~6 mouse melanoma cells were intravenously injected into C57BL/6J mice for two weeks,which is a recommended method for the establishment of melanoma lungmetastasis model.Part Ⅱ The mechanism of platelet membrane glycoprotein GPIbα in tumor metastasis【Objective】To investigate the mechanism of platelet membrane glycoprotein GPIbα in tumor metastasis.【Methods】1)The expression levels of GPIbα mRNA and protein in tumor cells were detected by Real-time PCR and Western Blot,respectively 2)The melanoma mouse model was constructed in GPIbα knockout mice.【Results】1)GPIbα was expressed on HCT116 cell line,but the expression level of GPIbα in HCT116 cell line was lower than that in human platelets.2)By tail vein injection with B16F10 cells,the number of lung metastases was significantly lower in GPIbα knockout mice than that in wild type mice.【Conclusion】Platelet membrane glycoprotein GPIbα promotes the metastasis of melanoma cells in mice.