TFu对肝门部胆管癌细胞系FRH-0201增殖和凋亡的影响

The effect of TFu on the apoptosis and proliferation of FRH-0201 cholangiocarcinoma cell line

目的：探讨N3-邻甲苯甲酰基-氟尿嘧啶（N3-toluyl—Flulorouractil TFu）对FRH-0201细胞的生物学效应及其作用机制，为胆管癌临床化疗提供参考。方法：光学显微镜、荧光显微镜观察FRH-0201细胞在TFu作用后形态学变化；集落形成实验检测TFu对FRH-0201细胞的增殖抑制作用；琼脂糖凝胶电泳检测DNA Ladder观察细胞的凋亡；流式细胞术检测细胞周期及细胞凋亡并与5-Fu进行比较；MTT试验比较TFu与5-Fu等常用化疗药物对FRH-0201细胞抑制率。结果：TFu能体外抑制FRH-0201细胞的增殖，其作用在0．016-4mmol／L范围内具有浓度依赖性及时间依赖性，同浓度比较抑制率高于5-Fu；流式细胞仪检测可见随作用时间延长，TFu及5-Fu组G1细胞期增多。S期细胞减少，细胞增殖被阻滞在G1期；细胞凋亡检测可见，随作用时间延长，TFu及5-Fu组较对照组细胞凋亡率增加，同时相TFu组凋亡率高于5-Fu组；化疗敏感性比较，TFu抑制FRH-0201细胞增殖作用不及阿霉素和顺铂，但明显优于5-Fu及泰素。联合用药：阿霉素、顺铂和TFu联合对FRH-0201细胞抑制率最高，达79．02％，优于阿霉素、顺铂、5-Fu联合。结论：TFu可明显抑制胆管癌细胞株FRH-0201细胞的增殖，该抑制作用可能通过阻滞细胞周期并进一步诱导细胞凋亡机制发生，同摩尔浓度TFu较5-Fu对FRH-0201细胞抑制作用强，TFu可作为胆管癌联合化疗的新药。

Objective： To investigate the effect of TFu on FRH-0201 cell and to test new drugs for the chemotherapy of cholargiocarcinoma. Methods ： Morphological chanses were closerved by light microscopy and fluorescence microscopy. The effect of TFu on FRH-0201 cell was examined by clone formation. Cell cycle and apoptosis were examined by flow-cytometric analysis. DNA ladder was used to detect apoptosls. To compare the drub susceptibility of TFU with 5-Fu and several other common drubs by MTT colorlmetrlc assay. Results ：TFu sianificantly Inhibited the proliferation and clone formation of FRH-0201 cell line within concentration of 0. 016-4mmol/ L. The Inhibition ratio was dependent on ascending concentration and time, and that Is hlaher than 5-Fu with the same concentration. The morphological character of apoptosis was observed. Flow cytometric analysis indicated that TFu Induced a G1 phase arrest and apoptosls more than 5-Fu.ADM,DDP and TFu were the sensitive clruas to FRH-020l cel line,and the conbination of them can reach the hlahest Inhibition ratio. Conclusion ： These results suggests that TFu can more slgnifisantly Inhibited the proliferation of FRH-0201 cell line than 5-Fu. The mechanism may be related to the cell cycle arrest In essoclatlon with apoptosls,whlch may provide a new approach for treating cholanaiocarcinoma.