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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 observe the inhibitory effect of interleukin-24 (IL-24) gene on the growth of Bel-7402 hepatoma cell line, and to provide a theoretical basis for gene therapy of hepatocellular carcinoma. Methods: The eukaryotic expression vector pIRES-IL-24 was transfected into Bel-7402 hepatoma cell line. The expression of IL-24 gene was detected by RT-PCR. The expression of IL-24 protein was detected by Western blotting and ELISA. The growth inhibition and killing effect of IL-24 on hepatocellular carcinoma cell were detected by MTT assay. The apoptosis of hepatoma cells was detected by flow cytometry cell cycle. Results: pIRES-IL-24 was able to express efficiently in Bel-7402. The concentration of IL-24 protein in cell culture supernatant was 124.1 ng / mL. IL-24 could significantly inhibit the growth of Bel-7402 hepatoma cells, and the inhibition rate was 46.3% on the 4th day after transfection, compared with the control group, P <0.05. IL-24 promoted the apoptosis of hepatocellular carcinoma cells, the apoptosis rate was 41.0%, compared with the control group, P <0.05. Cell cycle analysis showed that IL-24 blocks hepatocellular carcinoma cells in G2 / M phase. Conclusion: The recombinant plasmid pIRES-IL-24 can efficiently induce the expression of IL-24 gene in human hepatocellular carcinoma cells. It can kill the hepatoma Bel-7402 cell line, promote the cell proliferation and induce the apoptosis of tumor cells.