生存素反义核酸抑制肝癌细胞增殖及对5-FU的增敏作用

Survivin antisense oligodeoxynucleotides inhibits the proliferation of hepatocellular carcinoma cells and enhances the cell chemosensitivity to 5-Fu

目的探讨生存素反义核酸抑制肝癌细胞增殖及对5-FU的增敏作用。方法WST法、台盼蓝拒染法、克隆形成抑制实验检测聚乙烯亚胺携带生存素反义核酸(PEI-ASODN)对SMMC-7721细胞的增殖抑制作用。建立小鼠肝癌腋下移植瘤和腹水瘤模型,检测PEI-ASODN联合5-FU对肝癌移植瘤小鼠瘤重、瘤体积的改变,对肝癌腹水瘤小鼠平均存活天数的影响。结果不同浓度的PEI-ASODN作用于SMMC-7721细胞48h后,0.75μmol/L的PEI-ASODN分别作用细胞24、48、72和96h后,均能对细胞产生明显的抑制作用;PEI-ASODN对细胞的克隆形成也有显著的抑制作用,与细胞对照组相比有显著的统计学差异(P<0.01)。肝癌腋下移植瘤小鼠,各治疗组的瘤重、瘤体积均得到不同程度的抑制,其中以联合用药组(5-FU+PEI-ASODN)最明显,瘤重抑瘤率高达56.91%,瘤体积抑制率高达57.83%;肝癌腹水瘤小鼠模型,联合用药组生命延长率为94.09%,与生理盐水组相比,具有显著性差异(P<0.01)。结论聚乙烯亚胺携带生存素反义核酸(PEI-ASODN)可显著抑制肝癌细胞生长,并可提高荷肝癌小鼠对5-FU的敏感性。

Objective To investigate the inhibitory effect of survivin antisense oligodeoxynuleotides（ASODN） mediated by polyethylenimine（PEI） on the proliferation of hepatocelluar carcinoma SMMC-7721 cells,and assess its detect on the chemosensitivity of the cells to 5-FU.Methods The inhibitory effect of PEI-ASODN on SMMC-7721 cell proliferation was assessed using WST-8 test,trypan blue staining,and cell clone formation test.In mice bearing transplanted hepatocarcinoma and ascites tumor derived from H22 cells,5-FU combined with PEI-ASODN was administered,and the weight and volume of the subcutaneous tumors were measured to calculate the tumor inhibition rate,and the average survival time of the mice was calculated.Results Incubation of the cells with different concentrations of PEI-ASODN for 48 h significantly inhibited the cell proliferation as compared with the control group,but PEI or ASODN alone produced no significant inhibitory effects.At 24,48,72,96 h of incubation of the SMMC-7721 cells with 0.75 μmol/L PEI-ASODN,the cell proliferation was suppressed significantly,and incubation with PEI-ASODN at 0.25-0.75 μmol/L for 7 days resulted in significantly inhibited cell clone formation.No significant inhibition was detected in ASODN and PEI group.The tumor weight and volume were reduced in all the treated groups.The tumor inhibition rate was 56.91% and volume inhibition rate was 57.83% in 5-FU＋PEI-ASODN group,significantly different from those in the normal saline group（P〈0.01）.In mice bearing ascites tumor,the average survival time was 22.0 days in saline group,and 42.7 days 5-FU＋PEI-ASODN group.The life-prolongation rate of 5-FU＋PEI-ASODN was 94.09% when compared with the survival time in saline group.A cooperative effect was detected between 5-FU and PEI-ASODN.Conclusion PEI-ASODN complex can significantly inhibit the proliferation of hepatocarcinoma SMMC-7721 cells and enhance the chemosensitivity of the tumor cells to 5-FU.