三氧化二砷联合5-氟尿嘧啶对小鼠肝癌H_(22)细胞株的抑制作用

Inhibitory effects of combination of arsenic trioxide with 5-fluorouracil on mice liver cancer cell line H_(22) in vitro

目的:探讨三氧化二砷(As2O3)联合5-氟尿嘧啶(5-FU)对小鼠肝癌H22细胞株的抑制作用,为提高治疗肝癌的临床效果、降低毒性和克服耐药提供实验依据。方法:采用MTT法检测不同浓度(终浓度为0.5、1.0、2.0μg/mL)的As2O3、5-FU(终浓度为20 mg/L)和以上各浓度As2O3联合5-FU体外作用于小鼠肝癌H22细胞24、48、72 h后对细胞增殖的影响,并计算细胞生长抑制率和两药物相互作用系数(CDI);采用TUNEL荧光染色法检测细胞凋亡,观察各组细胞凋亡情况并计算凋亡率。结果:体外As2O3或5-FU单药对小鼠肝癌H22细胞有显著抑制增殖及诱导凋亡的作用,并呈一定的时间、浓度依赖性。As2O3联合5-FU对小鼠肝癌H22细胞的抑制增殖及诱导凋亡作用较同时间内相应单药组效果明显提高。计算两药CDI值,各浓度As2O3与5-Fu联合作用H22细胞24和48 h时,均为CDI<1,作用72 h时,均为CDI=1,说明两者在48 h内具有协同作用,72 h时为相加作用。结论:体外As2O3和5-FU对小鼠肝癌H22细胞有明显地抑制增殖和诱导凋亡作用,并且呈时间和剂量依赖性;联合应用较单药效果明显增强,在48 h内两药具有协同作用,72 h时两药作用相加。

Objective：To evaluate the inhibitory effects of arsenic trioxide （As2O3） combined with 5-fluorouracil （5-FU） on mice liver cancer cell line H22 in vitro,and to provide the experimental evidence for the clinical effects in treating liver cancer.Methods：As2O3 in different doses （0.5,1.0,2.0 μg/mL） and 5-FU （20 mg/L） were used to treat mice liver cancer cell line H22 for 24,48,72 h respectively alone or in combination,and then cells proliferation was assayed by MTT method,and to calculate the coefficient of them in interaction; TUNEL （Fluorescein） was utilized to detect the apoptosis of mice liver cancer cell line H22 treated by As2O3 in different doses （0.5,1.0,2.0 μg/mL） and 5-FU （20 mg/L） respectively alone or in combination in 24,48,72 h,calculating the apoptosis rate.Results：In vitro,As2O3 and 5-FU either combined or alone inhibited the proliferation of mice liver cancer cell line H22 and induced apoptosis significantly,depending on the time and consistency.And the inhibitory effects of combined groups were higher than that of alone groups significantly.The coefficient of the two drugs in interaction was synergistic （CDI＜1） in 24 h and 48 h,but in 72 h it was additive （CDI=1）.Conclusion：In vitro,As2O3 or 5-FU has significant effects of inhibiting proliferation and inducing apoptosis on mice liver cancer cell line H22 depending on its woking time and dosage.And the inhibitory effects of combined groups are higher than that of alone groups significantly.The coefficient of the two drugs in interaction is synergistic in 24 h and 48 h,but in 72 h it is additive.