熊胆粉对肝癌移植瘤裸鼠STAT3通路的影响  

Effect of Bear Bile Powder on STAT3 Pathway in Hepatocellular Carcinoma Xenograft

作　　者：赵锦燕[1] 刘丽雅[1] 沈阿灵[1] 林薇[1] 曹治云[1] 庄群川[1] 洪振丰[1] 

机构地区：[1]福建中医药大学中西医结合研究院,福州350122

出　　处：《中国中西医结合杂志》2014年第8期976-981,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：福建省科技厅重大专项基金资助项目(No.2010YZ0001-1)

摘　　要：目的观察熊胆粉对肝癌皮下移植瘤裸鼠信号转导与转录激活因子3(STAT3)通路及其下游靶基因的影响,探讨熊胆粉治疗肝癌的作用机制。方法采用人肝癌细胞株HepG_2构建裸鼠皮下移植瘤模型,待皮下移植瘤形成后,将裸鼠随机分为熊胆粉组和对照组,每天分别给予熊胆粉和生理盐水,连续给药3周,每周测量体重和瘤体积2次,给药结束,剥取瘤体称量瘤重及计算抑瘤率;采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)检测瘤组织的细胞凋亡情况;逆转录-聚合酶链反应(RT-PCR)检测Bcl2-associated X蛋白(Bax)、B细胞淋巴瘤/白血病-2(BcI-2)、周期蛋白依赖性蛋白激酶4(CDK4)、细胞周期蛋白D1(cyclinD1)mRNA表达;免疫组化测定磷酸化的信号转导与转录活化因子3(p-STAT3)、增殖细胞核抗原(PCNA)、Bax、Bcl-2、CDK4、cyclinD1蛋白表达。结果熊胆粉显著抑制肿瘤的体积和瘤重,与对照组比较,差异均有统计学意义(P<0.01)。TUNEL法结果显示熊胆粉显著促进肝癌细胞凋亡。RT-PCR结果显示熊胆粉促进Bax表达而抑制Bcl-2、CDK4、cycIinD1 mRNA表达。免疫组化结果显示熊胆粉促进Bax表达而抑制p-STAT3、PCNA、BcI-2、CDK4、cyclinD1蛋白表达。结论熊胆粉可以通过调控STAT3通路抑制肝癌细胞增殖,促进肝癌细胞凋亡。Objective To observe the effect of bear bile powder （BBP） on the STAT3 pathway and its downstream target genes of nude mice hepatocellular carcinoma （HCC） xenograft, and to ex- plore its mechanism for treating HCC. Methods The subcutaneous xenograft model was established u- sing HepG2 cells. When the subcutaneous transplanted tumor was formed, naked mice were randomly di- vided into two groups, the BBP group and the control group. Mice in the BBP group were administered with BBP by gastrogavage, once daily for 3 consecutive weeks, while mice in the control group were ad- ministered with normal saline by gastrogavage, once daily for 3 consecutive weeks. The body weight and the tumor volume were measured once per week. By the end of medication, the tumor weight was weighed and the tumor inhibition ratio calculated. The apoptosis of the tumor tissue was detected by TdT- mediated dUTP nick end labeling {TUNEL）. The expression of Bcl2-associated X protein （Bax）, B cell lymphoma/leukemina-2 （Bcl-2）, cyclin-dependent protein kinase （CDK4）, cyclinD1 were detected by re- verse transcription-polymerase chain reaction { RT-PCR）. The protein expression levels of signal trans- ducers and transcription activators 3 （p-STAT3）, proliferating cell nuclear antigen （PCNA）, Bax, Bcl-2, CDK4, and cyclinD1 were determined by immunohistochemistry. Results BBP could inhibit the tumor volume and tumor weight, showing statistical difference when compared with the control group （P 〈0.01 ）. Results of TUNEL showed that BBP could significantly induce the apoptosis of hepatoma car- cinoma cells. Results of RT-PCR showed that BBP could up-regulate the expression of Bax and down-reg-ulate mRNA expression of Bcl-2, CDK4, and cyclinDl. could up-regulate the expression of Bax and inhibit the CDK4, and cyclinDl. Conclusion BBP could induce the hibit their proliferation by regulating STAT3 pathway. mmunohistochemical results showed that BBP protein expression of p-STAr3, PCNA, Bcl-2, apoptosis of hepatoma carcin

关 键 词：熊胆粉 肝癌 信号转导与转录激活因子3 

分 类 号：R285[医药卫生—中药学]