外源性三磷酸腺苷对人食管癌Eca-109和肝癌SMMC-7721细胞增殖和周期的影响

Effects of extracellular ATP on apoptosis of human esophageal carcinoma Eca-109 and hepatoma SMMC-7721 cells

目的研究三磷酸腺苷（ATP）对于人食管鳞癌Eca-109细胞系和人肝癌SMMC-7721细胞系的抗增殖作用和作用机制。方法用MTT法测定肿瘤细胞的增殖，AO／EB双荧光染色法观察细胞形态学的改变，流式细胞仪定量检测细胞增殖周期、凋亡率及凋亡相关蛋白的改变。结果ATP在0．01～0．3mmol／L浓度范围内，可不同程度地抑制Eca-109和SMMC-7721细胞的增殖。细胞分别经0．3mmol／L的ATP处理72h后，少量SMMC-7721细胞形态出现了凋亡特征，而Eca-109细胞无明显凋亡特征；ATP0．03、0．1和0．3mmol／L分别处理细胞72h后，两株细胞的凋亡率及凋亡相关蛋白均无显著变化。ATP0．3mmol／L可使Eca-109细胞的G0／G1细胞数目显著增多，S期细胞显著减少，增殖指数显著降低。结论ATP通过增加G0／G1期细胞和减少S期细胞，抗食管癌Eca-109细胞增殖，此作用与诱导细胞凋亡无关。而ATP对肝癌SMMC-7721细胞的抗增殖作用与周期无关。

Objective To study the effects of adenosine triphosphate （ATP） on proliferation of human squamous esophageal carcinoma Eca-109 and hepatoma SMMC-7721 cells in vitro and the underlying mechanism. Methods MTT assay was used to determine the proliferation of tumor cells. The AO/EB double stained cells were observed under fluorescence microscope. The effects of ATP on the cell cycle, apoptotic rate and apoptosis-related protein were detected by flow cytometry. Results ATP showed inhibitory effects on Eca-109 and SMMC-7721 cells at the concentration of 0. 01 N 0. 3 mmol/L. Exposure to 0. 3 mmol/L ATP for 72 h, some of SMMC-7721 cells displayed morphological changes of apoptosis, but Eca-109 cells did not show the characteristics of apoptosis markedly. There was no significant change in the apoptotic rate and apoptosis-related protein of the two tumor cell lines treated with ATP 0.03, 0. 1 and 0. 3 mmol/L for 72 h respectively . The proportion of Eca-109 cells in G0/G1-phase of cell cycle was significantly increased, meanwhile the proportion of Eca-109 cells in S-phase and proliferation index value was significantly decreased by treatment with 0. 3 mmol/L ATP. Conclusion ATP inhibited Eca-109 cell proliferation by changing the distribution of cell cycle phase, and its mechanism might not related to apoptosis, but for SMMC-7721 cell, the inhibition of cell proliferation induced by ATP was not related to the change in cell cycle.