【摘要】 目的检测瓣状核酸内切酶1(flap endonuclease-1,FEN1)及增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在口腔癌相关成纤维细胞(carcinoma-associated fibroblasts,CAFs)中的表达水平,并分析两者表达水平的相关性。方法选取新鲜的口腔鳞状细胞癌组织块和因口腔颌面部整形手术切除的正常口腔黏膜组织块,采用组织块贴壁法培养原代口腔CAFs和正常成纤维细胞(normal fibroblasts,NFs)并行免疫细胞化学染色鉴定;并分成CAFs组和NFs组,分别采用Western blot和PCR检测口腔CAFs和NFs中FEN1、PCNA蛋白表达水平及m RNA水平,并对FEN1、PCNA的表达水平做相关性分析。结果成功培养并鉴定口腔CAFs和NFs各12株,FEN1、PCNA的m RNA和蛋白表达水平在口腔CAFs组均高于NFs组,但差异均无统计学意义(P>0.05);在口腔CAFs中,FEN1与PCNA在m RNA水平呈正向的强相关性(r=0.677,P=0.016),而两者的蛋白表达水平无相关性(P>0.05)。结论在口腔CAFs中FEN1和PCNA的m RNA和蛋白表达水平与NFs组相比均无统计学差异,但CAFs中的FEN1与PCNA二者在m RNA水平呈正向强相关,两种基因在口腔CAFs中的相互关系及调节机制尚需进一步研究。更多还原

【Abstract】 Objective To research the expression levels of FEN1 and PCNA in carcinoma-associated fibroblasts(CAFs) and analyze their correlation. Methods Fresh specimens of oral squamous cell carcinoma tissues and normal oral mucosal tissues excised during oral and maxillofacial plastic surgery were collected. Primary oral CAFs and normal fibroblasts(NFs) were obtained by tissue culture, identified by immunocytochemistry and divided into the CAF and NF groups. Western blot and quantitative real-time PCR were used to detect the protein and m RNA expression levels of both FEN1 and PCNA in the oral CAFs and NFs. The correlation between FEN1 and PCNA expression in oral CAFs was analyzed. Results Oral CAFs and oral NFs were successfully cultured and identified from 12 samples. Both the protein and m RNA expression levels of FEN1 and PCNA were higher in the oral CAFs than NFs, but there were no significant differences(P > 0.05). In the oral CAFs, the linear correlation coefficient between FEN1 and PCNA was 0.677(P = 0.016) at the m RNA level, indicating a strong positive correlation; however, at the protein level, no correlation was found(P > 0.05). Conclusion In primary cultured oral CAFs and NFs, there were no significant differences in the FEN1 and PCNA protein and m RNA expression levels. However, in the CAFs, the m RNA levels of FEN1 and PCNA had a strong positive correlation. The relationship and the regulatory mechanism of the two genes require further study.更多还原