Different responses of HepG2 subclones to low dose ethaselen  

低剂量乙烷硒啉对HepG2细胞亚株的影响不同（英文）

作　　者：张国州[1] 熊堃 马薇薇 徐伟 曾慧慧 

机构地区：[1]北京大学医学部药学院化学生物学系,天然药物及仿生药物国家重点实验室,北京100191

出　　处：《Journal of Chinese Pharmaceutical Sciences》2016年第4期302-309,共8页中国药学（英文版）

基　　金：National Natural Science Foundation(Grant No.81372266);National Science and Technology Major Project,People’s Republic of China(Grant No.2011zx09101-001-03)

摘　　要：As a synthesized antineoplastic organoselenium compound, ethaselen is known to induce apoptosis in tumor cells via dose-dependent thioredoxin reductase （TrxR） inhibition. Thioredoxin, the multifunctional biological substrate of TrxR, is then left in the oxidized state, which subsequently leads to intracellular accumulation of reactive oxygen species （ROS）, cell cycle arrest and/or apoptosis. However, the low dose effect of ethaselen remains largely unknown. Several subclones have been derived from HepG2 cells by using single cell or colony isolation. The low dose of ethaselen was defined as the drug concentration of retaining 〉90% HepG2 cells alive. The HepG2 cells were used as reference of its subclones （SM01, SM02 and SM03）, and the cell cycle transition, intracellular proteins change, colony formation and sphere growth were assayed in treatment of low dose ethaselen. HepG2 and its subclones differently responded to lethal dose of cisplatin or 5-fluorouracil. Low dose of ethaselen （1 μm） modulated the cell cycle transition at 12 h of treatment, but ceils were partially recovered at 24 h of treatment though some proteins were still affected. Low dose of ethaselen did not inhibit the small colony （diameter 〉 100 μm） formation and sphere growth of HepG2 and SM01. However, low dose of ethaselen could specifically inhibit the survival, large colony （diameter 〉500 μm） formation and sphere growth of SM03, although SM03 could be rapidly recovered from ethaselen-induced cell cycle check. HepG2 and its subclone cells could survive but respond differently to treatment of low dose ethaselen （1 μM）. Low dose of ethaselen could significantly inhibit a HepG2 subclone （SM03） in cell survival and colony growth.作为一种合成的、有抗肿瘤活性的有机硒,乙烷硒啉可通过抑制硫氧还蛋白还原酶,导致其生物学底物硫氧还蛋白处于氧化状态,随后升高的细胞内活性氧引发细胞周期阻滞或触发凋亡。乙烷硒啉的低剂量作用尚未被研究。本研究中乙烷硒啉低剂量(1μM)定义为保持90%以上细胞存活的浓度。从单个HepG2细胞增殖而来的亚株(SM01,SM02,SM03)被用于低剂量乙烷硒啉效应检测。考察了低剂量乙烷硒啉对HepG2及其亚株的细胞周期转变、蛋白表达、集落形成、细胞球生长变化的影响。HepG2及其亚株对致死剂量的CDDP和5-FU显示了不同的敏感性。低剂量乙烷硒啉处理12h后S期比例升高,可在随后的12 h内部分恢复,其细胞内蛋白p53,NF-κB等尚未完全恢复。低剂量乙烷硒啉未抑制较小的细胞集落(直径>100μm)形成,未影响HepG2和SM01的细胞球生长。低剂量乙烷硒啉可显著抑制SM03细胞存活、较大细胞集落(直径>500μm)形成和细胞球生长,尽管SM03细胞蛋白水平恢复较快。总体上,HepG2及其亚株对与化学物和低剂量乙烷硒啉的反应不同,低剂量乙烷硒啉可抑制HepG2亚株(SM03)的存活和生长。

关 键 词：Low dose ORGANOSELENIUM Ceil subcloning HETEROGENEITY Tumor cell 

分 类 号：R96[医药卫生—药理学]