神经突触核蛋白-γ过表达降低肝癌细胞对抗微管药物的敏感性  

Overexpression of synuclein-γ confers resistance to antimicrotubule drugs against human hepatoma cells

作　　者：程世翔[1,2] 张赛[2] 张豪[1] 宋丹青[1] 王宇萍[1] 李玉环[1] 游雪甫[1] 王跃明[1] 蒋建东[1] 

机构地区：[1]中国医学科学院、北京协和医学院医药生物技术研究所,北京100050 [2]中国人民武装警察部队医学院附属医院,天津300162

出　　处：《药学学报》2010年第6期724-729,共6页Acta Pharmaceutica Sinica

基　　金：高等学校全国优秀博士学位论文作者专项资金(200564);中央级公益性科研院所基本科研业务专项基金(IMBF200803)

摘　　要：肝癌是中国最常见的恶性肿瘤之一。目前,紫杉醇(paclitaxel,PTX)和长春新碱(vincristine,VCR)等抗微管药物是临床上肝癌化疗的常用药物,然而此类药物对患者的疗效差别很大。因此,发现影响抗微管药物作用的细胞内因子具有重要意义。本研究采用人肝癌细胞株HepG2稳定过表达synuclein-γ(SNCG)模型,研究SNCG是否影响肿瘤细胞对抗微管药物的敏感性。实时定量PCR(real-timeRT-PCR)和蛋白免疫印迹(Western blotting)法检测HepG2/SNCG细胞稳定高表达SNCG,与HepG2/Neo细胞相比,SNCG mRNA表达上调13.6倍,增殖活性增高(66±5)%。经PTX或VCR分别处理,SNCG过表达可降低抗微管药物诱导的细胞有丝分裂期阻滞,HepG2/SNCG细胞的G2/M期比例较HepG2/Neo细胞分别下降(19±2.9)%和(27±1.7)%;HepG2/SNCG、HepG2/Neo和HepG2细胞对PTX的有丝分裂指数分别为0.47、0.64和0.67,对VCR的有丝分裂指数分别为0.14、0.30和0.34。HepG2/SNCG细胞对PTX、VCR的敏感性显著降低,耐药指数分别为21和15。研究结果表明,SNCG在肝癌细胞过表达后可显著降低其对抗微管药物的敏感性,在肝癌化疗中SNCG是潜在的抗微管药物使用的生物标志物。Liver cancer is one of the most common neoplastic diseases with high mortality in China.Currently,antimicrotubule drugs such as paclitaxel（PTX） and vincristine（VCR）,are used as the common agents in the clinical chemotherapy for liver cancer.However,the responses of patients to these drugs vary markedly.Successful identification of intracellular factors influencing liver cancer＇s sensitivity to antimicrotubule drugs would be of great clinical importance.In this study,by engineering human hepatoma cell HepG2 to overexpress synuclein-γ（SNCG）,we investigated if SNCG is a molecular factor associated with the sensitivity to antimicrotubule drug treatment.Real-time RT-PCR and Western blotting assays showed SNCG was successfully overexpressed in HepG2/SNCG cells compared with HepG2/Neo cells.The overexpressed SNCG altered the proliferation activity in HepG2 cells,which was 66% higher than that of HepG2/Neo cells through MTT method.The overexpressed SNCG also reduced sensitivity of HepG2 cells to antimicrotubule drugs：after PTX or VCR treatment,the proportion of HepG2/SNCG cells in G2/M arrest was significantly lower than that in HepG2/Neo cells.Correspondingly,HepG2/SNCG cells showed significantly lower mitotic index than HepG2/Neo cells.Meanwhile,HepG2/SNCG cells showed higher resistance to PTX and VCR than HepG2/Neo cells,with resistance index 21 and 15 respectively.Our studies suggested that the overexpression of SNCG could confer resistance to antimicrotubule drugs in hepatoma cells;and it indicated that SNCG may be as a potential response marker for antimicrotubule drugs in liver cancer chemotherapy.

关 键 词：神经突触核蛋白-γ 抗微管药物 耐药性 肝癌 

分 类 号：R963[医药卫生—微生物与生化药学]