摘要
目的探讨以人骨髓间充质干细胞(hBMSCs)为基因治疗载体表达外源性IL-24对肺癌细胞A549增殖和凋亡的影响。方法分离培养hBMSCs,并通过流式细胞术鉴定。慢病毒介导IL-24基因转染hBMSCs,构建表达外源性基因IL-24的hBMSCs(hBMSCs-IL-24),qPCR及ELISA法检测hBMSCs-IL-24中IL-24基因mRNA及蛋白表达。CCK-8法检测hBMSCs-IL-24分泌的外源性IL-24对肺癌细胞A549增殖活性的影响。AnnexinV-PI检测hBMSCs-IL-24对A549细胞凋亡的影响。结果慢病毒介导的外源基因IL-24成功转染hBMSCs(hBMSCs-IL-24),其IL-24基因在mRNA及蛋白水平均有明显表达。hBMSCs-IL-24分泌的外源性IL-24蛋白显著抑制肺癌细胞A549的增殖并促进其凋亡。结论 hBMSCs-IL-24能够在mRNA及蛋白水平表达外源性IL-24基因,IL-24蛋白能显著抑制肺癌细胞A549的增殖,促进凋亡。
[ Objective ] To study the influence of human bone marrow mesenchymal stem cells （hBMSCs） infected by LV-IL-24 on the proliferation and apoptosis of A549 lung cancer cells. [Methods] The hBMSCs were isolated and cultured from human bone marrow and verified by flowcytometry. Recombinant lentiviral vectors harboring IL- 24 infected with hBMSCs to construct hBMSCs-IL-24 containing exogenous IL-24. Expression of IL-24 mRNA and IL-24 protein levels were detected by real-time quantitative PCR （qPCR） and ELISA. CCK-8 method was used to detect the effect of hBMSCs-IL-24 supernatant on the proliferation of A549 lung cancer cells. AnnexinV-PI method was used to detect the effect of hBMSCs-IL-24 on the apoptosis of A549 lung cancer cells. [ Results ] Exogenous IL-24 gene was successfully transfected into hBMSCs by recombinant lentivirus （hBMSCs-IL-24）, IL-24 expressed in hBMSCs-IL-24 at both mRNA and protein levels as detected by qPCR and ELISE. The exogenous IL-24 protein secreted from hBMSCs-IL-24 significantly inhibited the proliferation and promoted the apoptosis of A549 lung can-cer cells. [ Conclusions ] The hBMSCs-IL-24 can express IL-24 at mRNA and protein levels, IL-24 protein can significantly inhibit the proliferation and promote apoptosis of A549 lung cancer cells.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2014年第9期11-16,共6页
China Journal of Modern Medicine
基金
国家自然科学基金(No:81071909)
