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

Raf-1 is possibly related with the multidrug resistance in oral squamous cell carcinoma

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 Multidrug resistance is a major barrier for most patients with oral squamous cell carcinoma. In this study, we investigated the difference of the expression of Raf-1 mRNA and protein between multidrng resistant （KBV） and sensitive （KB） human oral squamous carcinoma cells to explore the effect of Raf-1 on the multidrug resistance in the human oral squamous carcinoma. Methods The changes of the mRNA and protein levels of Raf-1 in both KBV cells and KB cells were determined by RT-PCR, Western blotting assay and immunofluorescent method. By MTF and flow cytometry （FCM）, IC50 and apoptosis rates in both KBV and KB cells were detected. Results The findings indicated that expressions of Raf-1 mRNA and protein levels in KB cells were lower than those in KBV cells statistically （P 〈 0.01 ）. Immunofluorescent image showed that Raf-1 protein expression in KBV cells were much stronger than that in KB cells. The IC50 in KB cells was （21.33 ~ 2.25）μg/L and that in KBV cells was （352.68 ± 12.36）μg/L. The resistance index （RI） was 16.5.The apoptosis rate in KB cells was （98.8 ± 1.2） % and that in KBV cells was （23.5 ± 2.1 ）%. These results showed that differences in the chemosensitivity to VCR between KBV cells and KB cells were significant statistically （ P 〈 0.01 ）. Conclusion These studies suggested that Raf-1 might play an important role in the process of developing the muhidrug resistance in the human oral squamous carcinoma.