芹菜素对裸鼠肝癌皮下移植瘤抑制作用的研究

摘要

目的 探讨芹菜素(APG)对裸鼠肝癌皮下移植瘤的抑制作用及其相关机制.方法 建立人肝癌阿霉素(ADM)耐药细胞BEL-7402/ADM裸鼠皮下移植瘤模型,24只移植成功的裸鼠随机分为三组:对照组(ADM 3 mg/kg)、APG组(APG 50 mg/kg+ADM 3 mg/kg)和miR-101 mimics组(miR-101 mimics 50 mg/kg+ADM 3 mg/kg),每组8只.上述药物每3天注射1次,共7次.给药结束后处死裸鼠,切取移植瘤组织.免疫组化法检测Ki-67蛋白表达评价肿瘤细胞增殖,免疫蛋白印迹法检测肿瘤组织核转录因子红细胞系相关因子-2(Nrf2)蛋白的表达,qRT-PCR法测定移植瘤组织miR-101水平.结果 与对照组相比,APG和miR-101 mimics能显著抑制肿瘤生长(P＜0.05),抑制肿瘤细胞增殖(P＜ 0.05或P＜0.01),此外,APG和miR-101 mimics还能下调肿瘤组织Nrf2蛋白的表达(P＜0.01),上调miR-101表达(P＜ 0.05).结论 APG能有效抑制肝癌移植瘤生长,其机制可能与抑制肿瘤细胞增殖,上调miR-101表达和下调Nrf2表达有关.%Objective To investigate the effect of apigenin (APG) on hepatocellar carcinoma xenografts in nude mice and the possible molecular mechanism.Methods Models of nude mice were transplanted with BEL-7402/doxorubicin (ADM),and randomly divided into 3 groups,each group had 8 mice.The groups of mice were treated i.p.as follow:ADM (3 mg/kg,control group),APG+ADM (50 mg/kg+3 mg/kg,APG group),miR-101 mimics +ADM (50 mg/kg+3 mg/kg,miR-101 mimics group),every three days for a total of seven times.The volumes of tumor were measured.After the end of medication,the mice were sacrificed for measurement of tumor weights.Tumor cell proliferation was analyzed by Ki-67 immunostaining.Nuclear factor erythroid-2-related factor 2 (Nrf2) protein level was measured by Western blot.miR-101 level was detected by Real-time PCR.Results The volumes and weights of tumors in APG group and miR-101 mimics group were decreased compared with the control group (P ＜ 0.05).Compared to the control group,APG and miR-101 mimics resulted in inhibition of tumor cell proliferation (P ＜ 0.05 or P ＜ 0.01),inhibition of Nrf2 expression (P ＜ 0.01),and up-regulation of miR-101 levels (P ＜ 0.05).Conclusion APG can inhibit the growth of hepatocellar carcinoma,its mechanism may partly be to inhibit the expression of Ki-67 and Nrf2,up-regulate the level of miR-101.