硫利达嗪对肝癌干细胞的杀伤作用研究  

Killing Effects of Thioridazine on Liver Cancer Stem Cell

作　　者：孟树林 马步云[1] 张新敏[1] 葛云[1] 张蓉[1] 黄盼盼[1] 王毅刚[1] 

机构地区：[1]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

出　　处：《中国生物工程杂志》2015年第2期8-17,共10页China Biotechnology

基　　金：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

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。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 ceils 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 ceils were evaluated by MTY, 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.

关 键 词：THO 肝癌干细胞 磷酸化STAT3 CASPASE 凋亡 

分 类 号：Q819[生物学—生物工程]