rhIL-24蛋白诱导THP-1细胞分泌炎症性因子和肿瘤细胞凋亡作用的研究  

Induction effect of rhIL-24 protein on THP-1 cell secreting inflammatorycytokines and tumour cell apoptosis

作　　者：何涛君[1,2] 陆学东[1] 焦海春[2] 胡杰[2] 肖建华[2] 

机构地区：[1]广东医学院附属深圳市福田人民医院检验医学部,广东深圳518033 [2]湖南衡阳南华大学病原生物学研究所,湖南衡阳421001

出　　处：《中华肿瘤防治杂志》2009年第20期1546-1551,共6页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨rhIL-24蛋白诱导THP-1细胞分泌炎症性因子和肿瘤细胞凋亡的意义,为临床肿瘤的基因治疗奠定基础。方法:RT-PCR扩增人源性成熟肽IL-24基因,构建重组质粒pQE30/hIL-24,在大肠埃希菌M15中表达重组蛋白。SDS-PAGE和蛋白质印迹法对表达的产物鉴定,镍-琼脂糖凝胶层析柱纯化蛋白,尿素梯度透析法复性,ELISA法和FCM法检测rhIL-24蛋白诱导THP-1细胞分泌IL-6、TNF-α炎症性细胞因子和THP-1凋亡。结果:在大肠埃希菌中表达的rhIL-24蛋白与预期值相一致。纯化并复性后的rhIL-24蛋白能诱导THP-1分泌炎症性因子,蛋白浓度28μg/mL作用THP-1细胞,细胞因子的分泌量达到峰值,rhIL-24蛋白作用THP-1细胞18h,分泌量达到峰值。THP-1细胞随着rhIL-24浓度的增加凋亡率不断增加,28μg/mL达到峰值。结论:rhIL-24蛋白诱导THP-1分泌炎症性因子和细胞凋亡,说明rhIL-24诱导肿瘤细胞凋亡和免疫刺激抗肿瘤的协同效应得到验证。OBJECTIVE：To explore the significance of rhIL-24 protein induction THP-1 cell secretion inflammatory cytokines and tumor cell apoptosis.METHODS：The recombinant plasmid pQE30/hIL-24 was constructed,then it was identified by PCR,restriction enzyme analysis and sequence analysis.The engineering bacteria with recombinant plasmid pQE30-hIL24 which has been identified by sequence analysis were transformed into E.coli M15 expressing rhIL-24 protein.Then the biologic function of this protein was detected.hIL-24 gene was amplified by RT-PCR and the recombinant plasmid pQE30/hIL-24 was constructed.It was induced by IPTG and was identified by SDS-PAGE and Western-bolt.Human histomonocyte leukemic cells（THP-1 cells）were stimulated by rhIL-24,THP-1 cell secreting cytokines of TNF-α and IL-6 were tested by ELISA and apoptosis was tested by FCM.RESULTS：After purification and renaturation,rhIL-24 protein stimulated THP-1 cells to produce inflammatory cytokines TNF-α and IL-6,simultaneously this protein induced the apoptosis of THP-1 cells.It was indicated that inflammatory cytokines TNF-α and IL-6 secrete the maximizing volume after rhIL-24 protein 28 μg/mL acting on THP-1 cells for 18 h.THP-1 cells apoptosis reached the maximum after rhIL-24 protein 28 μg/mL acting on them.CONCLUSIONS：rhIL-24 protein can induce THP-1 cells secreting inflammatory cytokines and apoptosis.The synergistic effects of rhIL-24 protein on tumour cell apoptosis and immunostimulation anti-tumour have gained a tentative evidence.

关 键 词：rhIL-24蛋白 THP-1细胞 炎症趋化因子类 凋亡 

分 类 号：R73-3[医药卫生—肿瘤]