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
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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.Abstract: 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 14.21 μM, 10.56 μM, 10.39 μM; 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.
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Key words:
- Thyroid papillary carcinoma /
- Ursolic acid /
- Apoptosis /
- Cycle /
- Bcl-2 /
- Bax /
- Caspase-9
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[1]陈锡强, 程丽芳, 徐新刚, 等.熊果酸对斑马鱼血管生成及斑马鱼移植癌的抑制作用[J].中国药理学通报, 2015, 31 (7) :1004-1008. [2]付伦, 朱敏, 李招云, 等.熊果酸对肺癌细胞株A549及SPCA1细胞周期的抑制作用[J].医学研究杂志, 2015, 44 (5) :68-73. [3]张军辉, 严家芹, 陈小兵.熊果酸对Hep-2细胞增殖的影响及机制研究[J].中国现代医学杂志, 2015, 25 (12) :7-10. [4]庞姗姗, 朱悦, 周逸婵, 等.熊果酸通过ROS/AMPK/STAT3信号通路抑制胃癌细胞COX-2表达[J].江苏医药, 2015, 31 (7) :1004-1008. [5]岳军.熊果酸诱导HL-60细胞凋亡及抑制增殖与PTEN-Akt途径的相关性[J].中国医药科学, 2015, 5 (17) :31-33. [6] [6]PRASAD S, YADAV V R, SUNG B, et al.Ursolic acid inhibits the growth of human pancreatic cancer and enhances the antitumor potential of gemcitabine in an orthotopic mouse model through suppression of the inflammatory microenvironment[J].Oncotarget, 2016, 7 (11) :13182-13196 [7] [7]KASHYAP D, SHARMA A, TULI H S, et al.Ursolic acid and oleanolic acid:Pentacyclic terpenoids with promising anti-Inflammatory activities[J].Recent Patents on Inflammation&Allergy Drug Discovery, 2016, 10 (1) :21. [8] [8]YIE Y, ZHAO S, TANG Q, et al.Ursolic acid inhibited growth of hepatocellular carcinoma Hep G2 cells through AMPKα-mediated reduction of DNA methyltransferase[J].Molecular&Cellular Biochemistry, 2015, 402 (1-2) :63-74. [9] [9]WEN J H, WEI X H, SHENG X Y, et al.Effect of ursolic acid on breast cancer resistance protein-mediated transport of rosuvastatin in vivo and vitro[J].Chinese Medical Sciences Journal, 2015, 30 (4) :218-225. [10]董芬, 张彪, 单广良.中国甲状腺癌的流行现状和影响因素[J].中国癌症杂志, 2016, 26 (1) :47-52. [11] [11]COOPER D S, DOHERTY G M, HAUGEN B R, et al.Revised american thyroid association management guidelines for patients with thyroid nodules and differentiated thyroid cancer.[J].Thyroid Official Journal of the American Thyroid Association, 2016, 26 (1) :1. [12] [12]LEWINSKA A, ADAMCZYK-GROCHALA J, KWASNIEWICZ E, et al.Ursolic acid-mediated changes in glycolytic pathway promote cytotoxic autophagy and apoptosis in phenotypically different breast cancer cells[J].Apoptosis, 2017, 22 (6) :800-815. [13] [13]WU C C, CHENG C H, LEE Y H, et al.Ursolic acid triggers apoptosis in human osteosarcoma cells via caspase activation and the ERK1/2 MAPK pathway[J].Journal of Agricultural&Food Chemistry, 2016, 64 (21) :4220. [14] [14]CHUANG W L, LIN P Y, LIN H C, et al.The apoptotic effect of arsolic acid on SK-Hep-1 cells is regulated by the PI3K/Akt, p38 and JNK MAPK signaling pathways[J].Molecules, 2016, 21 (4) :460. [15] [15]ZHANG R X, LI Y, TIAN D D, et al.Ursolic acid inhibits proliferation and induces apoptosis by inactivating Wnt/β-catenin signaling in human osteosarcoma cells[J].International Journal of Oncology, 2016, 49 (5) :1973. -
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