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目的筛选小鼠黑色素瘤细胞中蛋白4.1家族的表达,探讨蛋白4.1对小鼠黑色素瘤细胞增殖的影响。方法以MEF、B16、B16-F10细胞基因组RNA和总蛋白为模板,经PCR法和Western blot筛选蛋白4.1在小鼠黑色素瘤细胞中的表达;通过分子克隆构建真核表达载体p EGFP-N1-EPB41L3并转染黑色素瘤细胞;检测表达蛋白4.1B后B16、B16-F10细胞增殖的变化。结果蛋白4.1B在黑色素瘤细胞中表达缺失;通过转染成功构建的真核表达载体p EGFP-N1-EPB41L3发现,表达蛋白4.1B的黑色素瘤细胞的增殖作用被显著性抑制。结论通过真核表达载体p EGFP-N1-EPB41L3转染表达蛋白4.1B可显著抑制B16和B16-F10黑色素瘤细胞的增殖。
Objective To screen the expression of protein 4.1 in mouse melanoma cells and investigate the effect of protein 4.1 on the proliferation of mouse melanoma cells. Methods The genomic RNA of MEF, B16 and B16-F10 cells and total protein were used as templates to screen the expression of protein 4.1 in mouse melanoma cells by PCR and Western blot. The eukaryotic expression vector p EGFP-N1- EPB41L3 were transfected into melanoma cells. The proliferation of B16 and B16-F10 cells was detected after the expression of protein 4.1B. Results The protein 4.1B was deleted in melanoma cells. The transfection of eukaryotic expression vector p EGFP-N1-EPB41L3 showed that the proliferation of melanoma cells expressing protein 4.1B was significantly inhibited. Conclusion The expression of protein 4.1B by eukaryotic expression vector p EGFP-N1-EPB41L3 can significantly inhibit the proliferation of B16 and B16-F10 melanoma cells.