三磷酸腺苷对人横纹肌肉瘤细胞系诱导分化作用的研究

ATP INDUCED DIFFERENTIATION OF HUMAN RHABDOMYOSARCOMA CELLS

为了探讨三磷酸腺苷（ＡＴＰ）对人横纹肌肉瘤细胞增殖和分化的影响，用ＡＴＰ作用于人横纹肌肉瘤细胞亚系（ＲＤＬ６）细胞，观察到ＡＴＰ可抑制ＲＤＬ６细胞的增殖，使其生长速度明显减慢，作用第５ｄ时增殖抑制率为８１％，流式光度术检测；观察到ＡＴＰ作用的ＲＤＬ６细胞Ｓ期的细胞数明显增多，说明细胞停滞在Ｓ期，用罗氏黄荧光染料传输法证实，ＡＴＰ有恢复ＲＤＬ６细胞间隙连接通讯功能的作用。用免疫荧光细胞化学方法观察到经ＡＴＰ处理后的ＲＤＬ６细胞间隙连接蛋白基因的表达明显增强。用肌专一蛋白荧光染色法也证明了ＡＴＰ促进ＲＤＬ６细胞的肌专一蛋白表达；出现肌丝肌管的分化。这些结果表明ＡＴＰ不仅抑制人横纹肌肉瘤细胞的增殖，而且诱导其分化，是有效的促分化性抗癌药物。本文讨论了增殖分化偶联与间隙连接蛋白基因（ｃｏｎｎｅｘｉｎ４３。

esults of experiments showed that ATP treatment inhibited cell proliferation and induced myogenic differentiation of RDL 6 cells which were from a single cell subclone of human rhabdomyosarcoma cell line RD. Through (1) determination of cell growth rate; (2) FCM of cell cycle; (3) examination of cell differentiation by muscle myosin heavy chain (MHC) immunofluorescence; (4) scrape loading and dye transfer techniques to detect gap junctional intercellular communication and (5) gap junction protein Cx43 immunocytochemistry to observe Cx43 gene expression, it was found that ATP not only inhibited cell proliferation, blocked RDL 6 cell cycle in S phase but also induced myogenic differentiation in RDL 6 cells. There was coupling relation existed between proliferation and differentiation of ATP treated RDL 6 cells, mean whilie the ATP treated RDL 6 cells showed increased gap junctional intercellular communication and enhanced Cx43 expression. These results indicated that enhanced functional expression of gap junction gene Cx43 play important roles in differentiation of the tumor cells.