骨髓间充质干细胞表达分泌型蛋白IFN-β/EGFP对前列腺癌PC-3细胞增殖的影响  

Effect of human mesenchymal stem cells expressing secreted protein IFN-β/EGFP on proliferation in prostate cancer cells line PC-3

作　　者：詹以安[1] 王共先[1] 傅斌[1] 

机构地区：[1]南昌大学第一附属医院泌尿外科研究所,南昌330006

出　　处：《第三军医大学学报》2012年第8期761-764,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30560153)~~

摘　　要：目的β干扰素(interferon-beta,IFN-β)/增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)真核质粒转染人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs),观察转染后的hMSCs(IFN-β-hMSCs)对前列腺癌细胞株PC-3增殖情况的影响。方法构建IFN-β/EGFP分泌型真核表达质粒pEGFP-N2-IFN-β,脂质体介导下转染hMSCs,使用荧光显微镜检测IFN-β/EGFP蛋白中绿色荧光的表达;ELISA法检测细胞上清中IFN-β的表达水平。将IFN-β-hMSCs与PC-3在Transwell培养体系中共培养,同时设立hMSCs共培养组、空质粒转染hMSCs共培养组、PC-3单独培养组为对照组,分别于共培养后第1、3、5天对PC-3进行台盼蓝染色,计数存活细胞数目。结果荧光显微镜下观察到IFN-β-hMSCs显示出绿色荧光,ELISA检测证实IFN-β-hMSCs的细胞上清中存在IFN-β的分泌表达。实验组PC-3细胞增殖受抑制,实验组与各对照组之间比较差异存在统计学意义(P<0.05)。结论分泌型蛋白IFN-β/EGFP在hMSCs中可获得表达且保持其各自的生物学特性,在Transwell共培养中可以有效抑制前列腺癌细胞增殖。Objective To determine the effect of interferon-~ （IFN-~）/ enhanced green fluorescent protein （EGFP）-expressing mesenchymal stem cells （IFN-13-hMSCs） on the proliferation of prostate cancer cell line PC-3. Methods The secreted eukaryotie expressed vector for recombinant secreted protein IFN-13/EGFP was constructed and then transfected into hMSCs by lipofeetamine. EGFP was detected by inverted fluorescence microscopy. The expression of IFN-~ in the medium of hMSCs-IFN-13 was detected by ELISA. IFN-13-hMSCs were incubated with prostate cancer cell line PC-3 in Transwell co-culture system, hMSCs, mock-hMSCs and PC-3 served as control cells. PC-3 cells were trypsinized and viable cells were counted respectively in 1,3, and 5 d using a hemocytometer after trypan blue staining. Results Fluorescence microscopy showed that the cells had positive fluorescent signals, and IFN-~ was detected in the culture medium of IFN-13-hMSCs. IFN-^-hM- SCs resulted in a significant decrease in PC-3 cells survival as compared with control groups （ P 〈 0.05 ）. Conclusion hMSCs expresses secreted protein IFN-~/EGFP successfully after gene transfeetion, and IFN-13- hMSCs cells inhibit growth significantly in the prostate cancer cells. IFN-~ and EGFP keep their individual characteristics with no interference with each other.

关 键 词：前列腺肿瘤 癌 Β干扰素 增强型绿色荧光蛋白 骨髓间充质干细胞 

分 类 号：R341[医药卫生—基础医学]