bFGF反义寡核苷酸增强鼠黑色素瘤B16细胞化疗药物敏感性研究

目的:研究bFGF反义硫代寡核苷酸增强肿瘤细胞对化疗药物敏感性作用。方法:设计、合成bFGF寡核苷酸,用聚乙烯亚胺(polyemyleneimine,PEI)介导bFGF反义硫代寡核苷酸转染入黑色素瘤B16细胞,MTT法检测bFGF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率;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;流式细胞仪分析bFGF反义硫代寡核苷酸诱导的细胞凋亡。结果:bFGF反义硫代寡核苷酸对B16细胞增殖的抑制率为64.8%,且呈剂量依赖效应。B16细胞中bFGF mRNA被bFGF反义硫代寡核苷酸显著降低,为对照细胞的57.9%,且bFGF反义硫代寡核苷酸诱导B16细胞凋亡,凋亡率为41.8%。bFGF反义硫代寡核苷酸转染能显著增强B16细胞对阿霉素、5-氟脲嘧啶及顺铂的敏感性,非特异性硫代寡核苷酸不影响阿霉素、5-氟脲嘧啶及顺铂抑制B16细胞增殖。结论:bFGF反义硫代寡核苷酸显著增强B16细胞的化疗敏感性,表明其可协同化疗药物用于治疗肿瘤。

Antisense Phosphorothioate Oligonucleotides Targeting bFGF Enhanced Chemosensitivity in Mouse Melanoma B16 Cells

Objective:To detect whether a specific antisense phosphorothioate oligonucleotide targeting bFGF enhance chemosensitivity of carcinomas.Methods:bFGF-specific antsense phosphorothioate oligonuleotides were transfected into mouse melanoma 1316 cell lines with jetPEI(polyethyleneimine).The relative cell proliferation was analyzed by MTT assay,bFGF mRNA level was evaluated by semi-quantitative RT-PCR,and apoptosis of the B 16 cells induced by antisense phosphorothioate oligonucleotides tar- geting bFGF was analyzed by flow cytometry.Cell proliferation was then analyzed after treatment with both bFGF-specific antsense phosphorothioate oligonuleotides and chemotherapeutic drugs by MTT assay.Results:bFGF-specific antisense phosphorothioate oligonuleotides inhibited the growth of B 16 cells by 64.8% in a dose and time-dependent manner,significantly reduced the bFGF mRNA level to 57.9% of control cells,and induced apoptosis by 41.8%.Compared to nonspecific phosphorothioate oligonuleotides,bFGF-spe- cific antisense phosphorothioate could significantly enhance chemosensitivity of B 16 cells to adriamycin(ADM),5-fluorouracil(5-FU), and cis-diaminodichloroptatin(DDP).Conclusion:bFGF-specific antisense phosphorothioate oligonucleotides have therapeutic effects on mouse melamoma B16 cells,and are a potential antitumor agent combined with chemotherapeutic drugs.