丹皮酚联合5-氟尿嘧啶对裸鼠人肝癌移植瘤的抑制作用及其机制

Anti-tumor Effects and Mechanisms of Paeonol Combined with 5-Fluorouracil Treatment for Nude Mice Transplanted Human Hepatic Carcinoma

目的探讨丹皮酚联合5-氟尿嘧啶对裸鼠人肝癌Bel-7404细胞皮下移植瘤裸鼠人肝癌移植瘤端粒酶活性及其hTERT表达的影响。方法通过建立裸鼠人肝癌Bel-7404细胞皮下移植瘤模型,将荷瘤裸鼠随机分为4组(每组5只):①模型对照(MC)组、②5-氟尿嘧啶治疗(5-Fu)组、③丹皮酚治疗(Pae)组、④5-氟尿嘧啶联合丹皮酚治疗(5-Fu+Pae)组。MC组经腹腔注射同体积0.9%氯化钠溶液;5-Fu组经腹腔注射5-Fu20 mg/kg;Pae组经腹腔注射Pae 100 mg/kg;5-Fu+Pae组经腹腔注射5-Fu20 mg/kg、Pae 100 mg/kg,隔日给药,连续给药5次。对比各组肿瘤的体积和质量差异,采用PCR-TRAP方法检测各组端粒酶活性,运用RT-PCR和Western blot法检测各组对端粒酶逆转录酶基因(human telomerasereverse transcriptase,hTERT)mRNA和蛋白表达变化。结果在采用5-Fu、Pae及联合用药干预后,肿瘤的体积和质量较模型对照组明显下降,其抑瘤率分别为45.32%、60.79%和80.22%。PCR和Western blot结果显示5-Fu、Pae及联合用药均能显著降低端粒酶活性和hTERT mRNA和蛋白的表达。结论 Pae具有协同5-Fu抗肿瘤的作用,其机制可能与降低端粒酶活性和hTERT的表达有关。

Objective To investigate the anti-tumor effects and mechanisms of paeonol（Pae） combined with 5-Fluorouracil（5-Fu） treatment for nude mice loaded human carcinoma of Bel-7404 cells.Methods The tumor model was established by subcutaneously injecting human carcinoma of Bel-7404 cells to right forearm armpits.Nude mice loaded Bel-7404 cells were randomly divided into 4 groups（5 in each group）：①Control group： equal volume NS intraperitoneal injection every two days for 10 days;②5-Fu group： 20 mg/kg,intraperitoneal injection every two days for 10 days;③Pae group：100 mg/kg,intraperitoneal injection every two days for 10 days;④Pae＋5-Fu group： Pae 100 mg/kg and 5-Fu 20 mg/kg intraperitoneal injection every two days for 10 days.We observed the tumor volumes and weights in each group.Telomerase activity was examined by PCR-TRAP.The expression of human telomerase reverse transcriptase（hTERT） mRNA and protein were analyzed by RT-PCR and Western blot,respectively.Results After the intervention of Pae,5-Fu,and their combination therapy,tumor volume and quality of decreased significantly compared with the control group,the tumor-inhibitory rates were 45.32%,60.79% and 80.22 %,respectively.The results of PCR and Western blot showed that Pae,5-Fu,and their combination therapy all could significantly suppress telomerase activity,and down-regulate mRNA and protein levels of hTERT.Conclusion Pae has a synergistic anti-tumor effect with 5-Fu and the mechanism may be related to reduced telomerase activity and hTERT expression.