Raf-1基因可能与口腔上皮癌细胞多药耐药性发生机制有关

• 作 者: 董岩 杨佩满 牛卫东
• 机 构: 大连医科大学口腔医学院,大连医科大学组织胚胎学教研室,
• 出 处: 解剖学报
• 发表日期: 2009年01期
• 关键词: 口腔上皮 Raf-1 翻译水平 耐药性 敏感细胞株 肿瘤细胞耐药 免疫印迹法 耐药株 细胞凋亡 多药耐药细胞株

目的通过检测Raf-1基因在口腔上皮癌多药耐药细胞株(KBV)及敏感细胞株(KB)中表达的差异,探讨Raf-1基因在口腔上皮癌细胞多药耐药性发生中的作用。方法体外培养口腔上皮癌多药耐药株及敏感株,通过RT-PCR技术及免疫荧光技术、免疫印迹技术检测Raf-1基因在KB及KBV细胞中转录及翻译水平的差异,通过MTT法及流式细胞检测技术(FCM)检测KB及KBV细胞对长春新碱(VCR)敏感度的差异。结果KB组细胞株在Raf-1基因转录和翻译水平均非常显著地低于KBV组(P<0.01);MTT法检测结果显示,KBV组细胞对长春新碱的耐药性是KB组的16.5倍;流式细胞术检测结果显示,KB组细胞凋亡率显著高于KBV组(P<0.01)。结论Raf-1基因可能参与口腔上皮癌耐药性的发生过程,成为引起肿瘤细胞耐药的重要原因之一。

Objective To detect the expression of Raf-1 gene in multidrug-resistant oral cancer cell lines (KBV) and sensitive cell lines (KB), and to investigate the role of Raf-1 in multidrug resistance of oral cancer cells effect. Methods The multidrug resistance and susceptible strains of oral epithelial carcinoma were cultured in vitro. The transcriptional and translational levels of Raf-1 gene in KB and KBV cells were detected by RT-PCR, immunofluorescence and immunoblotting. MTT and Flow cytometry (FCM) was used to detect the difference of sensitivity of vincristine (VCR) between KB and KBV cells. Results The transcriptional and translational levels of Raf-1 gene in KB group were significantly lower than those in KBV group (P <0.01). The results of MTT assay showed that the resistance of KBV cells to vincristine was 16.5 The results of flow cytometry showed that the apoptosis rate of KB group was significantly higher than that of KBV group (P <0.01). Conclusion Raf-1 gene may be involved in the pathogenesis of oral cancer drug resistance and become one of the important causes of tumor drug resistance.