携带p53基因增殖性腺病毒影响肝癌细胞对化疗药物的敏感性

Effect of replicating adenovirus harboring p53 gene on chemosensitivity of hepatocarcinoma cell line

目的:研究携带p53基因增殖性腺病毒CNHK600-p53载体能否增加肝癌细胞株对化疗药物的敏感性。方法:构建携带p53基因增殖性腺病毒CNHK600-p53载体,采用四甲基偶氮唑盐法(methylthiazolyl tetrazolium assay,MTT),观察并比较单用化疗药物[5-氟尿嘧啶(fluorouracil,5-Fu)、丝裂霉素(mitomycin,MMC)和表柔比星(epirubicin,EPI)]、单用CNHK600-p53和CNHK600-p53联合上述化疗药物对肝癌细胞株BEL-7404的杀伤效应。结果:肝癌细胞株BEL-7404在5-Fu浓度为100μg/ml时细胞抑制率为(47±3)%,MMC浓度为1μg/ml时细胞抑制率为(20±4)%,EPI浓度为2.5μg/ml时细胞抑制率为(73±2)%,联合CNHK600-p53(MOI=10)后上述浓度的化疗药物细胞抑制率分别增高为(59±4)%、(44±4)%和(86±2)%(率的U检验,P<0.01)。结论:携带p53基因的CNHK600-p53能提高肝癌细胞株BEL-7404对化疗药物的敏感性,CNHK600-p53联合化疗药物治疗可望开辟克服肝癌耐药的新途径。

Objective： To construct a replicating adenovirus vector CNHK600-p53 carrying p53 gene and investigate its effect on the chemosensitivity of hepatocarcinoma cell line. Methods： Chemotherapy of hepatocarcinoma cells BEL-7404 were carried out using Fluorouracil, Mitomycin, Epirubicin, CNHK600-p53, CNHK600-p53 ＋ Fluorouracil, CNHK600-p53 ＋ Mitomycin, and CNHK600-p53 ＋ Epirubicin, separately. MTT assay was used to evaluate the killing effects after therapy. Resuits： The inhibition rate on BEL-7404 cells was（47±3）% when using 100 μg/ml 5-Fu, was （20±4）% when using 1 μg/ml MMC, and was （73±2）% when using 2.5 μg/ml EPI. The inhibition rates of BEL-740 cells in CNHK600-p53（MOI= 10） ＋ Fluorouracil （100 μg/ml）, CNHK600-p53（MOI=10） ＋ Mitomycin （1 μg/ml）, and CNHK600-p53 （MOI= 10）＋ Epirubicin （2.5 μg/ml） groups were （59±4） %, （44±4） % and （86±2） %, respectively （P〈0.01）. Conclusion： Adenovirus CNHK600- p53 carrying p53 gene can enhance chemosensitivity of BELT404 hepatocarcinoma cell line. Gene therapy using adenovirus CNHK600-p53 in combination with chemotherapy may be a new strategy for hepatocarcinoma treatment.