微管相关蛋白2基因抑制恶性黑素瘤细胞增殖活性的研究

目的 探讨微管相关蛋白2(MAP-2)基因对恶性黑素瘤细胞生长的抑制作用.方法 扩增腺病毒-微管相关蛋白2(Ad-MAP-2)载体,空斑法测定Ad-MAP-2的滴度后,将Ad-MAP-2基因转染入鼠侵袭性恶性黑素瘤细胞B16C29细胞内.应用直接免疫荧光法测定细胞内MAP-2的表达,Ad-MAP-2对B16C29细胞生长的抑制作用,电镜观察细胞内微管形态,用流式细胞仪检测细胞凋亡.结果 Ad-MAP-2基因转染后,B16C29细胞胞体延伸出长而细的树枝状突起,近似于较成熟的色素细胞.电镜下见转染后细胞内微管含量增加,微管变长、变粗,延伸至树突,部分微管集合成束.转染后细胞增殖速度减缓,并出现凋亡.结论 MAP-2基因的表达可导致恶性黑素瘤细胞形态及微管状态的变化,抑制细胞增殖,诱导凋亡.

The inhibition of proliferation activity of melanoma cells by microtubule-associated protein-2

Objective To investigate the inhibition of melanoma cell lines by microtubule-associated protein 2 (MAP-2). Methods The adenoviral vector of MAP-2 (Ad-MAP-2) was constructed, then transfected into the mouse invasive melanoma cell line, B16C29. The expression of MAP-2 on the transfected cells was detected by direct immunofluorescence. Electron microscopy and flow cytometry were used to analyze the microtubule morphology and apoptosis of melanoma cells, respectively. Results The transfection was highly efficient. After the transfection, long and thin dendrites were observed on the melanoma cells, which looked like the mature melanocytes. The microtubules increased and became longer and thicker, extending to the dendrites. Some microtubules gathered to form bundles. The proliferation speed of the cells reduced, and apoptosis was observed. Conclusion The expression of MAP-2 can induce the changes in the morphology and microtubules of melanoma cells, inhibit the growth and induce the apoptosis of these cells.