反义核酸协同化疗药物抑制肝癌细胞的增殖

Synergism of an antisense oligodeoxynucleotides targeted to hTERT in combination with chemotherapeutic drugs on inhibiting the proliferation of HepG2 cells

目的 研究以端粒酶催化亚基hTERT基因为靶的反义硫代寡核苷酸(ASODN)分别与三种常用化疗药物顺铂(DDP)、5-氟尿嘧啶(5—FU)、阿霉素(ADM)联合用药抑制人肝癌细胞HepG2增殖的作用。方法 以脂质体为载体,将ASODN转染至HepG2中,用四氮唑盐(MTT)法染色并计算联用ASODN及化疗药物对HepG2增殖的抑制率,用SAS统计软件及金正均Q值法进行数据分析。结果 单用DDP、5-FU、ADM时其IC_(50)分别为1.07、4.15和0.29μg/ml,加入0.1μmol/L的ASODN,IC_(50)下降为0.25、1.52和0.12μg/ml,与单用化疗药的抑制效果相比有明显增强(F分别为66.92、25.96和8.56,P<0.001);Q值分析表明ASODN分别与中低浓度的DDP(≤1μg/ml)、5—FU(≤10μg/ml)、ADM(≤0.1μg/ml)联用有较好的协同作用(Q≥1.15)。结论 ASODN分别与化疗药物DDP、5—FU、ADM联合用药,均可显著增强抑制HepG2增殖的效果。

Objective To investigate the effect of a phosphorothioate antisense oligodeoxynucleotide 'ASODN' combined with cis-Diamminedichloroplatinum (DDP), 5-fluorouracil (5-FU) and adriamycin (ADM) respectively on inhibiting the proliferation of HepG2 cells. Methods A phosphorothioate antisense oligodeoxynucleotide (5'-ACTCACTCAGG CCTCAGACT-3') targeted to human telomerase reverse transcriptase (hTERT) mRNA, which named cantide, was synthesized. ASODN was transfected into HepG2 by lipofectin. And cell growth activity was evaluated by MTT assay. SAS software and Jin Zhengjun Method were used to evaluate the interaction of ASODN and these chemotherapeutic drugs. Results Combination treatments with 0.1 μmol/L ASODN reduced the IC50 of DDP, 5-FU and ADM from 1.07, 4.15 and 0.29 μg/ml to 0.25, 1.52 and 0.12 μg/ml respectively. The inhibitory ability of combination treatments on HepG2 cells was higher than that of these drugs alone (F = 66.92, 25.96, 8.56, P<0.001). And synergism (Q≥1.15) was observed at the lower concentration of DDP(≤1μg/ml), 5-FU (≤10μg/ml) and ADM(≤ 0.1 μg/ml) with combination of ASODN. Conclusion ASODN may enhance therapeutic effectiveness of chemotherapeutic drugs in human hepatocellular carcinoma cells.