表观遗传学药物对肝内胆管癌的作用  

Effects of epigenetic drugs in intrahepatic cholangiocarcinoma cells

作　　者：周文杰[1] 张锦前[2] 贺轲[1] 段小鹏[1] 黄睿[1] 夏正林[1] 何景亮[1] 向国安[1] 

机构地区：[1]南方医科大学第三临床医学院南方医科大学附属广东省第二人民医院普通外科,广州510317 [2]南方医科大学第三临床医学院南方医科大学附属广东省第二人民医院检验医学部,广州510317

出　　处：《中华实验外科杂志》2016年第3期662-665,共4页Chinese Journal of Experimental Surgery

基　　金：广东省自然科学基金（2015A030313725）

摘　　要：目的筛选肝内胆管癌敏感的表观遗传学药物并研究其作用机制。方法采用细胞活力实验筛选对RBE细胞敏感的药物；在多种细胞株上比较其作用；选择最敏感的药物探讨其机制：通过Giemsa染色观察肝内胆管癌细胞经药物处理后的形态学变化，用流式细胞技术检测细胞周期分布，联合流式细胞技术、Westernblot检测细胞的凋亡。结果RBE细胞分别经96种表观遗传学药物处理48h，27μmol／L浓度下22种药物对RBE细胞有抑制作用（细胞活力〈60％），组蛋白去乙酰化酶（HDAC）抑制剂有12种，3trmol／L浓度下仅5种药物对RBE细胞有抑制作用，HDAC抑制剂4种；其中，玉米原斑病菌毒素（HCtoxin）对多种肝内胆管癌细胞株的抑制作用最为突出；与对照组比较，HCtoxin处理CCLP-1细胞48h后，能引起包括小体凋亡等多种细胞形态变化，能将细胞周期阻滞在G。期（82．93％比48．51％），能通过非半胱氨酰天冬氨酸特异性蛋白酶．3（Caspase-3）依赖的凋亡途径诱导细胞凋亡。结论HDAC抑制剂，尤其是HCtoxin，通过多种方式对肝内胆管癌细胞产生显著的抑制作用。Objective Screening of epigenetic drugs sensitive to intrahepatic cholangiocarcinoma and exploring the mechanism. Methods Screening a 96 - epigenetic library in RBE cells through cell va- lidity assay and comparing the inhibition effect of these sensitive drugs on multiple cell lines ; Giemsa stai- ning was used to observe the cell morphological change and flow cytometry to check the cell cycle distribu- tion ; Morphological observation, flow cytometry technique and Western blotting were combined to check the cell apoptosis. Results Twenty two drugs from the 96 - epigenetic library had certain inhibition on RBE cells, including twelve histone deacetylase （HDAC） targeted species ; HC - toxin＇ s inhibition effect on four intrahepatic cholangiocarcinoma cell lines were the most obvious among the HDAC inhibitors＇ and were superior to that of gemcitabine; cells at G0/G~ phases in HC toxin treated group was much more than that in control group （82.93% vs. 48.51% ） and HC toxin brought about diverse of morphological chan- ges, apoptosis bodies and reduced mitosis included ; flow cytometry indicated intensively apoptosis inducted by HC toxin and it was not associated with Caspase - 3 depended pathways for the Caspase - 3 up - regula- ted in the trial groups tested by Western blotting was not remarkable in statistics. Conclusion HDAC in- hibitors, especially HC toxin, are of great potential in the treatment of intrahepatic cholangiocarcinoma.

关 键 词：肝内胆管癌 表观遗传学药物 组蛋白去乙酰化酶 玉米原斑病菌毒素 

分 类 号：R735.8[医药卫生—肿瘤]