熊果酸诱导甲状腺乳头状癌细胞TPC-1凋亡的实验  

Thyroid Papillary Carcinoma Cells TPC-1 Apoptosis Induced by Ursolic Acid

作　　者：华杰 阮凌玉 马定乾 左晓霜 向润清 方山丹 范源[2] HUA Jie;RUAN Ling-yu;MA Ding-qian;ZUO Xiao-shuang;FANG Shan-dan;FAN Yuan(College of Traditional Chinese Medicine;College of Clinical Medicine of Yunnan University of Traditional Chinese Medicine,Kunming Yunnan 650500,China)

机构地区：[1]云南中医学院中药学院,云南昆明650500 [2]云南中医学院临床医学院,云南昆明650500

出　　处：《昆明医科大学学报》2018年第1期10-15,共6页Journal of Kunming Medical University

基　　金：云南省应用基础研究计划项目-中医联合重点基金资助项目(2017FF117(-008))

摘　　要：目的研究熊果酸(ursolic acid UA)在体外对人甲状腺乳头状癌细胞株TPC-1增殖的抑制作用.方法采用不同浓度的UA干预细胞(对照组0μM,实验组3μM、6μM、12μM);MTT法观察同一时间不同浓度的UA和不同时间同一浓度的UA对TPC-1细胞生长情况的影响;流式细胞术(FCM)检测应用UA后TPC-1细胞凋亡情况及细胞周期分布情况;QRT-PCR检测UA干预后TPC-1细胞中Bcl-2、Bax、Caspase-9 m RNA的表达情况;Western blot检测UA干预后TPC-1细胞中Bcl-2、Bax、Caspase-9蛋白的表达情况.结果 MTT实验结果显示UA对TPC-1细胞增殖的抑制呈现出浓度和时间的依赖性,24 h、48 h、72 h的IC50分别为14.21μM、10.56μM、10.39μM;流式Annexin V-FITC/PI双染检测显示UA呈浓度依赖性诱导TPC-1细胞凋亡,且将TPC-1细胞的生长阻滞在S期;QRT-PCR实验发现,对照组与实验组均有Bcl-2、Bax、Caspase-9 m RNA的表达,UA呈浓度依赖性下调Bcl-2 m RNA的表达,上调Bax和Caspase-9 m RNA的表达;Western blot结果显示,对照组与实验组均有Bcl-2、Bax、Caspase-9蛋白的表达,UA呈浓度依赖性下调Bcl-2的表达,上调Bax和Caspase-9的表达.结论熊果酸可显著抑制人甲状腺乳头状癌TPC-1细胞的增殖并诱导其凋亡,为甲状腺癌治疗提供一定思路.Objective To study the inhibitory effect of ursolic acid on the proliferation of human papillary thyroid carcinoma cell line TPC-1 in vitro.Method TPC-1 cells were treated with different concentrations of ursolic acid(control group:0μM,experimental group:3μM,6μM,12μM);MTT assay was used to observe the effect of the growth of TPC-1 cells on different concentrations of ursolic acid at the same time;Apoptosis and cell cycle distribution of TPC-1 cells were treated with ursolic acid by flow cytometry;The expression of Bcl-2,Bax and Caspase-9 mRNA in TPC-1 cells were treated with ursolic acid by QRT-PCR;The expression of Bcl-2,Bax and Caspase-9 protein in TPC-1 cells were treated with ursolic acid by Western blot.Results MTT assay showed that ursolic acid inhibited the proliferation of TPC-1 cells in a concentration and time-dependent manner,and the IC50 at 24 h,48 h and 72 h was 14.21μM,10.56μM,10.39μM;Flow cytometry showed that ursolic acid inhibited the apoptosis of TPC-1 cells in a concentration-dependent manner,and the growth of TPC-1 cells was arrested in S phase;QRT-PCR showed that Bcl-2,Bax and Caspase-9 mRNA were expressed in the control and experimental groups,ursolic acid inhibited the expression of Bcl-2 mRNA in a concentration-dependent manner and up-regulated the expression of Bax and Caspase-9 mRNA;Western blot results showed that Bcl-2,Bax and Caspase-9 were expressed in the control and experimental groups,ursolic acid inhibited the expression of Bcl-2 protein in a concentration-dependent manner and up-regulated the expression of Bax protein and Caspase-9 protein.Conclusion Ursolic acid can significantly inhibit the proliferation and induce apoptosis of human papillary thyroid TPC-1 cells,providing some ideas for the treatment of thyroid cancer.

关 键 词：甲状腺乳头状癌 熊果酸 凋亡 周期 BCL-2 BAX CASPASE-9 

分 类 号：R736.1[医药卫生—肿瘤]