Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2015, Vol. 35 Issue (2): 8-17    DOI: 10.13523/j.cb.20150202
研究报告     
硫利达嗪对肝癌干细胞的杀伤作用研究
孟树林, 马步云, 张新敏, 葛云, 张蓉, 黄盼盼, 王毅刚
浙江理工大学生命科学学院新元医学与生物技术研究所 杭州 310018
Killing Effects of Thioridazine on Liver Cancer Stem Cell
MENG Shu-lin, MA Bu-yun, ZHANG Xin-min, GE Yun, ZHANG Rong, HUANG Pan-pan, WANG Yi-gang
School of Life Sciences, Xinyuan Institute of Medicine and Biotechnology, Zhejiang Sci-Tech University, Hangzhou 310018, China
 全文: PDF(1861 KB)   HTML
摘要:

硫利达嗪(Thioridazine,THO)在临床上通常用于治疗精神类疾病;近年来,研究发现THO对肿瘤细胞具有杀伤效果,但其对肝癌干细胞的杀伤作用还未曾有报道。肿瘤干细胞在肿瘤的转移、复发及耐药性方面起着十分重要的作用。利用体外悬浮培养富集肿瘤干细胞并检测药物THO对其杀伤效果,并以此评价THO对肿瘤生长的体外抑制效应。通过检测体外悬浮培养肝癌干细胞在肿瘤干细胞相关因子表达、耐药性及细胞周期等方面因素,显示在一定程度上其具备肿瘤干细胞样特征,磷酸化STAT3、NANOG和XIAP表达显著上调,而Albumin表达下调;进一步运用MTT、Western blotting和细胞流式等实验验证了THO对肝癌干细胞具有较强的杀伤效果并能诱导caspase依赖的细胞凋亡,而对分化的肝癌细胞影响较弱;此外,THO和化疗药物盐酸阿霉素(DOX)的联合使用显著增强了其对肝癌干细胞和分化的肝癌细胞的杀伤作用。因此,该结果首次显示THO对肝癌干细胞具有较强的杀伤能力,可能为今后肝癌的临床治疗带来新的希望。

关键词: THO肝癌干细胞磷酸化STAT3Caspase凋亡    
Abstract:

Thioridazine (THO) was originally used to treat psychotic patients. Recently, THO was shown to inhibit the growth of many cancer cell lines.But its killing effect on liver cancer stem cells have not yet been reported. Cancer stem cells play an important role in tumor metastasis, recurrence and drug resistance. The suspension culture was used to enrich cancer stem cells in vitro to test killing effect of THO on liver cancer stem cell. Liver cancer cells that cultured in suspension medium are detected similarly to liver cancer stem cells in many aspects, including embryonic transcription factors dependence, drug resistance and cell cycle arrest;which show that it significantly upregulated the expression of phosphorylated STAT3, NANOG and XIAP,and decreased expression of albumin proteins.The cytotoxicity of THO on liver cancer stem cells were evaluated by MTT, Western blotting and flow cytometry assay. The results indicate that THO can significantly inhibit the proliferation of liver cancer stem cells and induce caspase-dependent apoptosis;but weakly to liver cancer cells. However, the combination of THO and doxorubicin hydrochloride(DOX) could more significantly inhibit the proliferation of liver cancer stem cells and caner cells similarly. The results show that THO has a specific strong killing effect on liver cancer stem cells, and may bring new hope for future clinical treatment of liver cance.

Key words: THO    Liver cancer stem cells    Phosphorylated STAT3    Caspase    Apoptosis
收稿日期: 2014-11-18 出版日期: 2015-02-25
ZTFLH:  Q819  
基金资助:

国家自然科学基金(81272687)、国家"863 "计划(2012AA020806)、浙江省自然科学基金(LZ13H160004)、浙江省科技厅公益性技术应用研究计划(2014C33275)资助项目

通讯作者: 王毅刚     E-mail: Wangyigang43@163.com
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  

引用本文:

孟树林, 马步云, 张新敏, 葛云, 张蓉, 黄盼盼, 王毅刚. 硫利达嗪对肝癌干细胞的杀伤作用研究[J]. 中国生物工程杂志, 2015, 35(2): 8-17.

MENG Shu-lin, MA Bu-yun, ZHANG Xin-min, GE Yun, ZHANG Rong, HUANG Pan-pan, WANG Yi-gang. Killing Effects of Thioridazine on Liver Cancer Stem Cell. China Biotechnology, 2015, 35(2): 8-17.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150202        https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I2/8


[1] Siegel R, Ma J, Zou Z, et al. Cancer statistics, 2014. CA,2014,64(1):9-29.

[2] He G, Dhar D, Nakagawa H, et al. Identification of liver cancer progenitors whose malignant progression depends on autocrine IL-6 signaling,Cell,2013,155(2):384-396.

[3] 吴孟超,沈锋. 肝癌研究的现状和展望. 国外医学肿瘤学分册,2000, 27(1):17. Wu M C, Shen F. The present situation and prospects of research on liver cancer. Foreign Medical Sciences(cancer section), 2000, 27 (1): 17.

[4] Bonnet D, Dick J E. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nature Medicine,1997,3(7):730-737.

[5] Al-Hajj M, Wicha M S, Benito-Hernandez A, et al. Prospective identification of tumorigenic breast cancer cells. Proceedings of the National Academy of Sciences, 2003, 100(7):3983-3988.

[6] O'Brien C A, Pollett A, Gallinger S, et al. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature,2006,445(7123):106-110.

[7] Singh S K, Hawkins C, Clarke I D, et al. Identification of human brain tumour initiating cells. Nature,2004, 432(7015):396-401.

[8] Reya T, Morrison S J, Clarke M F, et al. Stem cells, cancer, and cancer stem cells. Nature,2001, 414(6859):105-111.

[9] Visvader J E, Lindeman G J. Cancer stem cells: current status and evolving complexities. Cell Stem Cell,2012,10(6):717-728.

[10] Yamashita T, Wang X W. Cancer stem cells in the development of liver cancer. The Journal of Clinical Investigation,2013,123(5):1911-1918.

[11] Huang P, Qiu J, Li B, et al. Role of Sox2 and Oct4 in predicting survival of hepatocellular carcinoma patients after hepatectomy. Clinical Biochemistry,2011,44(8):582-589.

[12] Shan J, Shen J, Liu L, et al. Nanog regulates self-renewal of cancer stem cells through the insulin-like growth factor pathway in human hepatocellular carcinoma. Hepatology (Baltimore, Md),2012, 56(3):1004-1014.

[13] Singh S K, Clarke I D, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors. Cancer Research,2003, 63(18):5821-5828.

[14] Kai K, Nagano O, Sugihara E, et al. Maintenance of HCT116 colon cancer cell line conforms to a stochastic model but not a cancer stem cell model. Cancer Science,2009,100(12):2275-2282.

[15] Tsuchiya A, Heike T, Fujino H, et al. Long-term extensive expansion of mouse hepatic stem/progenitor cells in a novel serum-free culture system. Gastroenterology,2005,128(7):2089-2104.

[16] Collura A, Marisa L, Trojan D, et al. Extensive characterization of sphere models established from colorectal cancer cell lines. Cellular and Molecular Life Sciences,2013, 70(4):729-742.

[17] Qiu X, Wang Z, Li Y, et al. Characterization of sphere-forming cells with stem-like properties from the small cell lung cancer cell line H446. Cancer Letters,2012,323(2):161-170.

[18] Zhang L, Jiao M, Li L, et al. Tumorspheres derived from prostate cancer cells possess chemoresistant and cancer stem cell properties. Journal of Cancer Research and Clinical Oncology,2012,138(4):675-686.

[19] Alimperti S, Lei P, Wen Y, et al. Serum-free spheroid suspension culture maintains mesenchymal stem cell proliferation and differentiation potential. Biotechnology Progress,2014,30(4):974-983.

[20] Reilly J, Ayis S, Ferrier I, et al. Thioridazine and sudden unexplained death in psychiatric in-patients. The British Journal of Psychiatry, 2002, 180(6):515-522.

[21] Huq Z U, Investigators R G. A trial of low doses of risperidone in the treatment of patients with first-episode schizophrenia, schizophreniform disorder, or schizoaffective disorder. Journal of Clinical Psychopharmacology, 2004, 24(2):220-224.

[22] Potkin S G, Thyrum P T, Alva G, et al. The safety and pharmacokinetics of quetiapine when coadministered with haloperidol, risperidone, or thioridazine. Journal of Clinical Psychopharmacology, 2002, 22(2):121-130.

[23] Choi A R, Kim J H, Yoon S. Thioridazine specifically sensitizes drug-resistant cancer cells through highly increase in apoptosis and P-gp inhibition. Tumor Biology, 2014,35(10):9831-9838.

[24] Kang S, Dong S M, Kim B R, et al. Thioridazine induces apoptosis by targeting the PI3K/Akt/mTOR pathway in cervical and endometrial cancer cells. Apoptosis: An International Journal on Programmed Cell Death, 2012, 17(9):989-997.

[25] Sachlos E, Risueño R M, Laronde S, et al. Identification of drugs including a dopamine receptor antagonist that selectively target cancer stem cells. Cell, 2012, 149(6):1284-1297.

[26] Cao L, Zhou Y, Zhai B, et al. Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines. BMC Gastroenterology, 2011, 11(1):71.

[27] Lee T K W, Castilho A, Cheung V C H, et al. CD24+ Liver tumor-initiating cells drive self-renewal and tumor initiation through STAT3-mediated NANOG regulation. Cell Stem Cell, 2011, 9(1):50-63.

[28] Bilim V, Kasahara T, Hara N, et al. Role of XIAP in the malignant phenotype of transitional cell cancer (TCC) and therapeutic activity of XIAP antisense oligonucleotides against multidrug-resistant TCC in vitro. International Journal of Cancer, 2003, 103(1):29-37.

[29] Yu Z, Pestell T G, Lisanti M P, et al. Cancer stem cells. The International Journal of Biochemistry & Cell Biology, 2012, 44(12):2144-2151.

[30] Hanahan D, Weinberg R A. Hallmarks of cancer: the next generation. Cell, 2011, 144(5):646-674.

[31] Yang J, van Oosten A L, Theunissen T W, et al. Stat3 activation is limiting for reprogramming to ground state pluripotency. Cell Stem Cell, 2010, 7(3):319-328.

[32] Chen W, Shen X, Xia X, et al. NSC 74859-mediated inhibition of STAT3 enhances the anti-proliferative activity of cetuximab in hepatocellular carcinoma. Liver International,2012,32(1):70-77.

[33] Sun C, Sun L, Jiang K, et al. NANOG promotes liver cancer cell invasion by inducing epithelial-mesenchymal transition through NODAL/SMAD3 signaling pathway. Int J Biochem Cell Biol, 2013, 45(6):1099-1108.

[34] Silva J, Nichols J, Theunissen T W, et al. Nanog is the gateway to the pluripotent ground state, Cell, 2009, 138(4):722-737.

[35] Johansson H, Simonsson S. Core transcription factors, Oct4, Sox2 and Nanog, individually form complexes with nucleophosmin (Npm1) to control embryonic stem (ES) cell fate determination. Aging-Us, 2010,2(11):815-822.

[1] 陶守松,任广明,尹荣华,杨晓明,马文兵,葛志强. 敲低去泛素化酶USP13抑制K562细胞的增殖*[J]. 中国生物工程杂志, 2021, 41(5): 1-7.
[2] 段阳阳,张凤亭,成江,石瑾,杨娟,李海宁. SIRT2抑制对MPP+诱导的帕金森病细胞模型凋亡和线粒体动态平衡的影响*[J]. 中国生物工程杂志, 2021, 41(4): 1-8.
[3] 郭利成,曹雪玮,傅龙云,王富军,赵健. 一种用于药物蛋白亲和纯化和跨膜转运的双功能标签的开发 *[J]. 中国生物工程杂志, 2020, 40(6): 40-52.
[4] 朱永朝,陶金,任萌萌,熊燃,何亚琴,周瑜,卢震辉,杜勇,杨芝红. 自噬抑制肿瘤坏死因子α诱导人胎盘胎儿来源间充质干细胞发生凋亡 *[J]. 中国生物工程杂志, 2019, 39(9): 62-67.
[5] 刘叶,潘玥,郑魏,胡晶. miR-186-5p在酒精诱导的心肌细胞中高表达并通过靶基因XIAP调控细胞凋亡水平 *[J]. 中国生物工程杂志, 2019, 39(5): 53-62.
[6] 汪路,杨丽媛,唐雨婷,陶瑶,雷力,敬一佩,蒋雪坷,张伶. 干扰PKM2对人白血病细胞增殖和凋亡的影响及潜在机制 *[J]. 中国生物工程杂志, 2019, 39(3): 13-20.
[7] 黄翔,杨杰,何佩彦,吴志慧,曾慧兰,王新宁,蒋建伟. 白花地胆草单体EM-12诱导2774-C10细胞G1/S期阻滞及细胞凋亡的分子机制研究[J]. 中国生物工程杂志, 2018, 38(4): 17-23.
[8] 代立婷, 吴忠南, 黄翔, 杨杰, 曾慧兰, 王国才, 蒋建伟. 卤地菊乙醇提取物W40单体诱导GLC-82细胞凋亡的分子机制研究[J]. 中国生物工程杂志, 2017, 37(8): 1-7.
[9] 徐安健, 李艳萌, 李斯文, 乌姗娜, 张蓓, 黄坚. PHP14沉默对肺癌细胞凋亡的影响及其机制[J]. 中国生物工程杂志, 2017, 37(7): 12-17.
[10] 李艳伟, 马义, 韩磊, 肖兴, 党诗莹, 文涛, 王得华, 范志勇. Fas凋亡抑制分子FAIM 1表达缺失诱发单纯性肥胖的初步研究[J]. 中国生物工程杂志, 2017, 37(6): 37-42.
[11] 白欣艳, 温丽敏, 王玉晶, 王海龙, 解军, 郭睿. ANKRD49通过上调Bcl-xL的表达抑制UV诱导GC-1细胞的凋亡[J]. 中国生物工程杂志, 2017, 37(4): 40-47.
[12] 冀君, 朱晨晨, 许鑫, 刘晓, 冷超粮, 史鸿飞, 姚伦广, 阚云超. 鸡贫血病毒凋亡素基因的可溶性融合表达及抗肿瘤活性分析[J]. 中国生物工程杂志, 2017, 37(2): 26-32.
[13] 李振华, 李翠平, 张相强, 代立婷, 唐梦思, 王国才, 蒋建伟, 曹明溶. EM-3通过Stat3通路诱导鼻咽癌细胞凋亡和G2/M期阻滞并降低SP细胞比例[J]. 中国生物工程杂志, 2016, 36(3): 1-10.
[14] 万春红, 张志, 李圣纳, 彭以元, 许亮国. TRAF7的研究进展[J]. 中国生物工程杂志, 2016, 36(3): 93-101.
[15] 张英敏, 赵娜, 李勇芳, 孟凡秀, 张琪, 高然朋, 张悦红, 于保锋, 郭睿, 王海龙, 解军, 徐钧. PBI-SUR-TK载体靶向介导HSV-TK自杀基因诱导肝癌细胞凋亡[J]. 中国生物工程杂志, 2016, 36(2): 16-21.