pIRES-IL-24对Bel-7402肝癌细胞抑制的实验研究

Inhibition effect of pIRES-IL-24 on Bel-7402 cells

目的:观察白细胞介素-24(IL-24)基因对肝癌细胞系Bel-7402生长的抑制作用,为肝癌的基因治疗提供理论基础。方法:将真核分泌表达载体pIRES-IL-24转染肝癌细胞系Bel-7402。RT-PCR检测IL-24基因的表达,蛋白质印迹法及ELISA检测IL-24蛋白的表达,MTT法检测IL-24对肝癌细胞的生长抑制和杀伤作用,流式细胞仪检测细胞的凋亡和细胞周期。结果:pIRES-IL-24能够在Bel-7402中高效表达。细胞培养上清液中IL-24蛋白表达浓度为124.1 ng/mL。IL-24能明显抑制Bel-7402肝癌细胞的生长,转染后第4天抑制率为46.3%,与对照组比较,P<0.05。IL-24促进肝癌细胞的凋亡,凋亡率41.0%,与对照组比较,P<0.05。细胞周期分析显示,IL-24阻滞肝癌细胞在G2/M期。结论:重组表达载体pIRES-IL-24介导IL-24基因在人肝癌细胞中高效表达,可杀伤肝癌细胞Bel-7402,促进细胞增殖阻滞及诱导肿瘤细胞凋亡。

OBJECTIVE：To investigate the inhibitory effect and anti-cancer mechanism of pIRES-IL-24 mediated IL-24 gene on hepatocarcinoma Bel-7402 cells and gene therapy of liver cancer theory.METHODS： The eukaryotic expression vector pIRES-IL-24 was transfected into hepatoma cell line Bel-7402.IL-24 gene expression was detected by RT-PCR,IL-24 protein expression was detected by Western and ELISA.IL-24 was assayed with MTT on hepatoma cell growth inhibition and cytotoxicity.The apoptosis and cell cycle were detected with FCM（flow cytometry）.RESULTS： pIRES-IL-24 efficiently expressed in Bel-7402 cells.In cell culture supernatant,IL-24 protein expression was detected,the concentration of IL-24 protein was 124.1 ng/mL.And IL-24 inhibited the Bel-7402 liver cancer cell growth.On the fourth day after transfection,the inhibition rate was 46.3% compared with the control group（P0.05）.IL-24 for liver cancer cell apoptosis,the apoptosis rate was 41.0%,compared with the control group,（P0.05）.The cell cycle analysis showed that liver IL-24 and block cancer cells were in G2/M phase.CONCLUSION： Recombinant expression vector pIRES-IL-24 mediated IL-24 gene is highly expressed in human liver cancer cells,and it could kill liver cancer cells Bel-7402,promote cell proliferation arrest and induce tumor cell apoptosis.