人黑色素瘤细胞侵袭转移恶性表型获得的分子学基础

Molecular basis of the malignant phenotype of the invasive transfer of the human melanoma cell

目的 探讨具有不同潜在转移能力的黑色素瘤细胞恶性表型的演进及其侵袭转移的机制。方法 采用细胞培养 ,p5 3蛋白免疫组化染色 ,PCR- SSCP检测 ,流式细胞术(PCM) ,蛋白酶活性分析 (Zymography)及裸鼠体内异种接种实验。结果 人黑色素瘤细胞中存在有 p5 3基因的突变 ,瘤细胞表面 6 7KD L N- R的荧光阳性率和全部细胞的平均荧光强度大小顺序为 WM45 1>WM983A>WM1341B>WM35。金属蛋白酶 (MMPs)的产生情况为 :早期 WM35不产生 MMPs;WM1341B仅产生 72 KDa(MMP- 2 ) ,不产生 92 KDa(MMP- 9) ;进展期 WM983A和远处转移瘤株 WM45 1均产生 72 KDa和 92 KDa。WM983A和 WM45 1也可见在裸鼠体内形成明显的转移瘤。结论 p5 3基因突变 ,MMPs的产生和细胞表面 6 7KD L N-

Objective To discuss the development and the invasive transfer mechanism.Methods Cell culture method, p53 protenin immunohistochemistry, PCR SSCP examination, zymography, PCM and heterogenic inouclation test inside the nude mice were adopted. Results p53 genetic mutation existed in human melanoma cells. The fluorescent positive rate of 67KD LN R on the surface of the melanoma cell and the average flurescent intensity are repectively WM451>WM983A>WM1341B>WM35.Early WM35 did not produce MMPs; WM1341B only produced 72KDa(MMP 2),not 92KDa(MMP 9);Both WM983A at progressive stage and distal transfering tumor WM45 produced 72KDa and 92KDa. WM983A and WM451 were also seen to form evident transfering tumor inside the naked mice.Conclusion p53 genetic mutation, the production of MMPs and the high indication of 67KD LN R on the celluar surface are the key factors for the human melanoma cell to obtain invasive transferring abililty.