慢病毒介导hIL-24基因对瘢痕疙瘩成纤维细胞增殖、迁移及侵袭性的影响

Effect of lentivirus-mediated hlL-24 gene on proliferation, migration and invasion of keloid fibroblasts

目的 研究人白细胞介素-24（human interleukin-24,hlL-24）基因对体外培养的人瘢痕疙瘩成纤维细胞（keloid fibroblasts,KF）增殖、迁移、侵袭性的影响.方法 将携带hIL-24及绿色荧光蛋白的慢病毒载体（LV-hlL-24-GFP,KF-hIL-24组）及空载体（LV-GFP,KF-NC组）感染KF细胞,并设空白对照组（KF组）,应用实时定量PCR和Western blot分别检测hlL-24 mRNA及蛋白的表达;MTT法检测KF细胞增殖情况;流式细胞仪法测定KF细胞周期;划痕实验和Trans-well小室实验检测KF细胞的体外迁移和侵袭能力.结果 hIL-24慢病毒载体感染KF细胞96 h后,KF-hIL-24组hIL-24 mRNA和蛋白的表达量明显高于KF-NC组和KF组,差异有统计学意义（P ＜0.01）;MTT结果显示,KF-hIL-24组吸光度A值明显降低,与KF-NC组比较,差异有统计学意义（P＜0.05）;细胞周期结果显示, hIL-24基因阻滞KF细胞在G1期[（75.40±2.10）％]、S期[（4.96±1.60）％]和G2期[（0.01 ±0.01）％]细胞比例下降,与KF-NC组比较,差异有统计学意义（P＜0.01）;划痕实验和Trans-well实验结果显示,hIL-24慢病毒载体能够抑制KF细胞的体外迁移和侵袭能力.结论 hlL-24慢病毒载体能够抑制KF细胞增殖、细胞周期进程、迁移及侵袭能力.

Objective To investigate the effect of hIL-24 gene on proliferation,migration and invasion activity of human keloid fibroblasts （KFs）.Methods hIL-24 gene was cloned into lentivirus vector,then the lentivirus particles expressing hlL-24 were infected into KF cells.Real-time PCR and Western blot were performed to examine the expression of hIL-24 in lentivirus infected cells.The growth ability was detected by MTT assay.The cell cycle was analyzed by flow cytometry,The invasion and migration were detected by matrigel invasion assay and wound healing assay.Results Comparing to controls group and KF-NC group,the expression levels of hIL-24 mRNA and protein were both significantly up-regulated after 4 days of hIL-24 lentivims infection.Comparing with the KF-NC group,MTT assay showed that the A490 of KF-hIL-24 group was down-regulated after lentivims infection （P 〈 0.05）.Comparing with the KF-NC group,Cell cycle test revealed hIL-24 gene could block KF cells in G1 [（75.40 ±2.10）%],the proportion of KF cells was decreased in S phase [（4.96 ± 1.60）%] and G2 phase [（0.01 ± 0.01） %].After KF cells were infected（P 〈 0.01）.Transfection of hIL-24 lentivirus inhibited the migration and invasion activity of KF cells.Conclusion Lentivirus-mediated hIL-24 gene efficiently inhibits proliferation,cell cycle progression,migration and invasion activity of KF cells.