bFGF反义硫代寡核苷酸增强人喉鳞癌Hep2细胞的化疗敏感性  

Antisense phosphorothioate oligonucleotide targeting bFGF enhances chemosensitivity of human laryngeal squamous carcinoma cells

作　　者：黄红亮[1] 王宏[1] 向军俭[1] 唐勇[1] 邓宁[2] 杨红宇[2] 

机构地区：[1]暨南大学分子免疫学与抗体工程中心,广州510632 [2]暨南大学 分子免疫学与抗体工程中心,广州510632

出　　处：《中国肿瘤生物治疗杂志》2007年第5期461-465,共5页Chinese Journal of Cancer Biotherapy

基　　金：中国博士后科学基金(No.20060390735);广东省生物工程药物重点实验室基金(No.51206001);Supported by the Postdoctoral Science Foundation of China(No.20060390735);Foundation for key Laboratory of Bioengineering Drug of Guangdong Province (No.51206001)

摘　　要：目的:研究bFGF反义硫代寡核苷酸增强人喉鳞癌Hep2细胞对多柔比星、氟脲嘧啶及顺铂的敏感性。方法:设计、合成bFGF反义脱氧寡核苷酸(AS),用聚乙烯亚胺(jetPEI)介导bFGF反义脱氧寡核苷酸转染入Hep2细胞;半定量RT- PCR测定bFGF反义硫代寡核苷酸转染后细胞中bFGF mRNA水平;免疫细胞化学法检测Hep2细胞转染前后bEGF的表达水平;FACS分析bFGF反义硫代寡核苷酸诱导细胞的凋亡;MTT法检测bEGF反义硫代寡核苷酸及其与化疗药物联合处理后的细胞增殖。结果:bFGF反义硫代寡核苷酸呈剂量与时间依赖抑制Hep2细胞增殖,最高抑制率为25.5%;被转染细胞bFGF mRNA和蛋白的表达分别降低52.0%和41.1%,细胞凋亡率为20.5%;bFGF反义硫代寡核苷酸协同多柔比星、氟脲嘧啶及顺铂作用Hep2,使3种药物的IC_(50)分别降低75.5%、83.5%及65.4%。结论:bFGF反义硫代寡核苷酸协同化疗药物增强Hep2细胞对化疗药物敏感性,将为人喉鳞癌的生物和化学药物治疗增添新途径。Objective： To study the enhancing effect of bFGF-targeted antisense phosphorothioate oligonucleotide （APO） on the chemosensitivity of human laryngeal squamous carcinoma cell line Hep2 to Doxorubicin, 5-Fluorouracil, and Cisplatin. Methods： bFGF-specific APO was designed, constructed and transfected into Hep2 cells with jetPEI （polyethyleneimine）. Expression of bFGF mRNA was evaluated by semi-quantitative RT-PCR after transfection ; immuno- cytochemical method was used to examine the expression of bEGF expression before and after transfection of Hep2 ; the induction of cell apoptosis was analyzed by flow cytometry ; cell proliferation was then analyzed by MTT assay after treatment with bFGF-specific APO or chemotherapeutic drugs, or a combination of both. Results： bFGF-specific APO inhibited the growth of Hep2 cells in a dose- and time-dependent manner, with the peak inhibitory rate being 25.5%. The expression of bFGF mRNA and protein decreased by 52.0% and 41.1%, respectively. The apoptosis rate of Hep2 cells was 20.5% after transfection, bFGF-specific APO reduced the 50% inhibitory concentration of Doxorubicin, 5-Fluorouracil, and Cisplatin in Hep2 cells by 75.5%, 83.5% and 65.4%, respectively. Conclusion： bFGF-specific APO can enhance the chemosensitivity of Hep2 cells, which paves a new way for potential biologic chemotherapy of laryngeal squamous carcinoma.

关 键 词：碱性成纤维细胞生长因子 喉鳞癌 反义硫代寡核苷酸 化疗药物 化疗敏感性 

分 类 号：R730.5[医药卫生—肿瘤] R739.6[医药卫生—临床医学]