摘要
目的:探讨全反式维甲酸(ATRA)对肝癌细胞HepG2的诱导分化作用及侵袭迁移的影响.方法:MTT法测定ATRA对肝癌细胞HepG2的抑制作用;平皿集落形成法检测ATRA对HepG2细胞锚定依赖性生长作用;ELISA法检测ATRA作用前后HepG2 AFP分泌量的变化;RT-PCR检测ATRA对HepG2细胞MMP-9及Nanog的mRNA表达的影响;Western blot检测ATRA对HepG2细胞MMP-9及Nanog蛋白表达的影响;Transwell及划痕实验检测其对侵袭及迁移能力的影响.结果:MTT法显示ATRA抑制体外培养人肝癌细胞HepG2细胞生长,呈剂量和时间依赖性;ELISA显示ATRA诱导HepG2细胞分化和凋亡,使代表肝细胞恶变的AFP分泌量明显下降(P<0.05);平皿克隆形成实验结果表明未经ATRA处理的HepG2可形成克隆,但经ATRA刺激后,其形成克隆变小且数目明显减少;ATRA呈时间及浓度依赖性下调Nanog mRNA及蛋白的表达,并在24h呈浓度依赖性下调MMP-9 mRNA及蛋白表达;Transwell实验和划痕实验结果显示ATRA能抑制HepG2细胞的侵袭和迁移能力.结论:ATRA可诱导HepG2细胞分化,降低HepG2的侵袭迁移能力,并可下调肿瘤干细胞分化调节基因Nanog的表达从而达到治疗肿瘤的目的.
AIM： To investigate the effect of all-trans retinoic acid （ATRA） on the differentiation, invasion and metastasis of liver cancer HepG2 cells. METHODS： After HepG2 cells were treated with different concentrations of ATRA, the proliferation of HepG2 cells was evaluated by MTT assay; anchorage-dependent growth was evaluated by colony formation assay; AFP secretion was determined by ELISA; the transcription levels of Nanog and MMP-9 were assessed by RT-PCR, and their protein levels were assessed by Western blot; and cell invasion and migration were evaluated by scratch test and transwell assay. RESULTS： ATRA suppressed the proliferation and anchorage-dependent growth of HepG2 in a doseand time-dependent manner. ATRA induced cell differentiation and decreased AFP secretion in HepG2 cells （both P 0.05）. Treatment with ATRA down-regulated the mRNA andprotein levels of Nanog and MMP-9 （within 24 hours） in a doseand time-dependent manner. In addition, ATRA could inhibit the invasion and metastasis of HepG2 cells. CONCLUSION： ATRA may induce cell differentiation, reduce cell invasion and migration and down-regulate the levels of Nanog in HepG2 cells.
出处
《世界华人消化杂志》
CAS
北大核心
2011年第33期3381-3389,共9页
World Chinese Journal of Digestology
关键词
全反式维甲酸
肝癌
增殖
分化
侵袭
迁移
All-trans retinoic acid
Liver cancer
Proliferation
Differentiation
Invasion
Migration
