ATP对人不死化成纤维细胞增殖及其细胞膜蛋白表达的影响

目的：探讨ATP对不死化人成纤维细胞增殖及其细胞膜蛋白表达的影响。方法：将正常人TIG-7和OUMS-36细胞株，不死化人KMST-6和SUSM-1细胞株在不同浓度的ATP、ADP、AMP条件下分别进行24和96 h的常规细胞培养，观察存活细胞数目，DNA的合成和³²P]-ATP标记膜蛋白的表达。结果：培养96 h后,0.4 mmol/L ATP对不死化细胞KMST-6的抑制率为77%且DNA合成明显地被抑制，而正常OUMS-36细胞抑制率为41%且DNA合成无明显改变。在1 mmol/L ATP时，多数KMST-6细胞发生死亡，而正常OUMS-36细胞的增殖无明显影响。当ADP、AMP和腺苷或磷酸处理的细胞，仅有ADP处理的不死化细胞存活数目减少（P<0.01）。与正常细胞比较，不死化细胞30 kD、31 kD、33 kD和40 kD的³²P]-ATP标记膜蛋白呈高表达。结论：ATP对不死化人成纤维细胞的增殖有明显的抑制作用，并且使磷酸化细胞膜蛋白表达增高。

Effect of ATP on proliferation and membrane protein expressionini mmortalized human fibroblasts

AIM: To investigate the effect of ATP on proliferationand membrane protein expression in immortalized human fibroblasts. METHODS: Applying the normal humanfi broblast cell line TIG-7 and OUMS-36, the immortalized human fibroblast cell line KMST-6 and SUSM-1 to culture for 24 hours and 96 hours with the different concentrations of ATP, ADP and AMP, respectively. The number of cell alive, DNA synthesis and the expressionof ³²P]-ATP labeled proteins in the membrane fraction were also observed. RESULTS: After 96 hours culture, the proliferation inhibition in immortalized cells exposed to 0.4 mmol/L ATP was 77%, while that of normal OUMS-36 cells treated withthe same concentration of ATP was 41%; the DNA synthesis of the immortalized cells exposed to 0.4 mmol/L ATP was greater suppressed than that of normal OUMS-36 cells. ADP was the only reagent tested thatreduced the number of cells significantly(P<0.01). When the immortalized cells exposed to ADP, AMP, adenosi ne, and phosphoric acid. In the presence of 1 mmol/L ATP, many KMST-6 cells died, but normal OUMS-36 cellsdid not. Compared with normal cells, the expresson of ³²P]-ATP labeled 30 kD, 31 kD, 33 kD and 40 kD proteins were markedly higher in the immortalized cells. CONCLUSION: ATP inhibited the proliferation and increased membrane protein expression in immortalized human fibroblasts.