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.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
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)
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
basic fibroblast growth factor
laryngeal squamous cell carcinoma
antisense phosphorothioate oligonucleotides
chemotherapeutic drugs
chemo sensitivity
