转染白细胞介素-12基因的脐带间充质干细胞对胃腺癌细胞的影响  

Effects of umbilical cord mesenchymal stem cells transfected with interleukin-12 gene on gastric carcinoma cells

作　　者：王久香 康柏会 司宁宁 赵芳菲 刘晓智[2] 

机构地区：[1]河北省滦平县中医院内科,068250 [2]天津市第五中心医院中心实验室,300450

出　　处：《广东医学》2015年第13期1987-1990,共4页Guangdong Medical Journal

基　　金：天津市滨海新区卫生局科技项目(编号:2013BWKY001)

摘　　要：目的探讨转染白细胞介素-12(IL-12)基因的脐带间充质干细胞(UCMSCs)对胃腺癌细胞的影响。方法原代分离培养和鉴定人UCMSCs;利用重组构建的IL-12和绿色荧光蛋白(GFP)融合表达的慢病毒载体(LV-IL-12-GFP)感染UCMSCs,酶联免疫法(ELISA)检测细胞中IL-12的表达。用转染了LV-IL-12-GFP的UCMSCs与胃腺癌细胞SGC-7901共培养,Transwell双室培养体系检测胃腺癌细胞SGC-7901的侵袭能力,原位细胞凋亡方法检测胃腺癌细胞SGC-7901的凋亡情况,Cylinder方法检测48 h后转染了LV-IL-12-GFP的UCMSCs向胃腺癌细胞SGC-7901的迁移能力。结果成功分离得到UCMSCs;ELISA结果显示,经IL-12基因转染的UCMSCs可稳定表达持续高水平的IL-12;Transwell结果显示,与转染了LV-IL-12-GFP的UCMSCs共培养的胃腺癌细胞SGC-7901的侵袭能力明显下降(P<0.05),凋亡率明显增加(P<0.05);Cylinder柱结果显示,转染了LV-IL-12-GFP的UCMSCs向胃腺癌细胞SGC-7901的定向迁移能力明显下降(P<0.05)。结论 UCMSCs可作为IL-12基因载体靶向追踪胃腺癌细胞,发挥肿瘤治疗作用。Objective To investigate the effects of umbilical cord mesenchymal stem cells（ UCMSCs） transfected with interleukin- 12（ IL- 12） gene on gastric carcinoma cells. Methods The isolated humanized UCMSCs were cultured and identified. UCMSCs were infected by the lentiviral vector（ LV- IL- 12- GFP） harboring the IL- 12 and green fluorescence protein（ GFP） genes. The expression level of IL- 12 was measured by enzyme- linked immunosorbent assays（ ELISA）. Gastric carcinoma cell line SGC- 7901 cells were co- cultured with UCMSCs transfected with LV- IL-12- GFP. The invasion ability of SGC- 7901 was monitored by the transwell dual chamber culture system,and the apoptosis of SGC- 7901 was detected by the in situ method. The migration ability of UCMSCs transfected with LV- IL- 12-GFP towards SGC- 7901 was monitored by Cylinder 48 hours later. Results UCMSCs were successfully isolated. The ELISA results showed the infected UCMSCs stably expressed a sustained high level of IL- 12. The transwell results presented the obviously reduced invasion ability and the increased apoptosis rate of SGC- 7901 co- cultured with UCMSCs transfected with LV- IL- 12- GFP（ P〈0. 05）. The Cylinder data revealed the obviously decreased ability of directional migration towards SGC- 7901 of UCMSCs transfected with LV- IL- 12- GFP（ P〈0. 05）. Conclusion UCMSCs can be used in the targeting therapy of gastric carcinoma cell as a vector of IL- 12.

关 键 词：胃癌 间充质干细胞 基因治疗 白细胞介素-12 

分 类 号：R722.12[医药卫生—儿科]