鸟氨酸脱羧酶基因反义RNA对肝癌细胞HepG2的影响

Effects of Antisense RNA of Ornithine Decarboxylase on HepG2 Hepatoma Cells

目的:探讨鸟氨酸脱羧酶(ornithine decarboxylase,ODC)基因反义RNA对肝癌细胞HepG2的影响。方法:构建ODC反义RNA的真核表达质粒,将此质粒转染HepG2细胞后,RT-PCR和Western印迹法筛选ODC表达抑制的细胞株。以此细胞株为模型,分析ODC反义RNA对细胞生长、细胞周期和对抗癌药物米托蒽醌敏感性的影响。结果:成功构建ODC反义RNA真核表达载体并获得稳定低表达ODC的肝癌细胞株Hr1。与对照细胞相比,ODC低表达引起HepG2细胞生长抑制,72h生长抑制率为31%;流式细胞术检测细胞周期发现,Hr1G1期细胞数(56.2%)显著性高于对照(48.2%),而S期细胞(25.5%)则显著性低于对照(34.9%),提示ODC低表达导致G1期阻滞;用米托蒽醌(100μg/L)处理两种细胞后发现,Hr1对药物的敏感性显著性高于对照细胞,处理48h后药物对HepG2和Hr1的抑制率分别是33.4%和60.6%,72h后的抑制率分别是60.8%和83.8%。结论:ODC反义RNA能抑制肝癌HepG2细胞生长,在抗肿瘤治疗中具有潜在的临应用价值。

Objective：To investigate the effects of antisense RNA of ornithine decarboxylase（ODC） on HepG2 cells.Method：The eukaryotic expression plasmid for ODC antisense RNA was constructed.The plasmid was then transfected into HepG2 human hepatoma cells.The cell line with ODC expression inhibition was selected by RT-PCR and Western blotting assay.Using this cell line as a model,the effects of ODC antisense RNA on cell growth,cell cycle and cell sensitive to antitumor drug were determined.Result：The plasmid for ODC antisense RNA was constructed successfully and a HepG2 cell line（Hr1） with stable low-expression of ODC was obtained.Comparing with the control cells,low-expression of ODC in Hr1 cell line resulted in growth inhibition,the inhibition rate at 72h was 31%.Flow-cytomitry assay shown that the cell number of G1 phase in Hr1（56.2%） was higher than that in the control cells（48.2%）,but the cell number of S phase in Hr1（25.5%） was lower than that in the control cells（34.9%）,which suggested that ODC low-expression resulted in cell cycle arrest at G1 phase.ODC low-expression increased cell sensitivity to antitumor drug mitoxantrone.When HepG2 and Hr1 were treated by 100μg/L mitoxantrone,inhibition rates were 33.4% and 60.6% respectively at 48h,and up to 60.8% and 83.8% respectively at 72h.Conclusion：Antisense RNA of ODC can inhibit growth of HepG2 cells and has potential value in the anti-tumor therapy.