骨肉瘤血清标志蛋白筛选及对RNA聚合酶Ⅲ多肽F的初步生物信息学分析

Screening of serum marker proteins in osteosarcoma and preliminary bioinformatic analysis on POLR3F

目的筛选潜在的骨肉瘤血清蛋白标志物，对候选标志基因RNA聚合酶Ⅲ多肽F（POLR3F）进行初步生物信息学分析。方法分别利用人类基因组芯片和表面增强激光解析电离飞行时间质谱技术（SELDI-TOFMS）筛选骨肉瘤细胞株以及骨肉瘤患者血清中差异表达的基因和标志蛋白。运用Link—test统计学方法对比基因：巷片和SELDI—TOFMS数据，寻找骨肉瘤的血清诊断标志蛋白。用MATLAB软件对POLR3F的启动子和3’-UTR区域进行生物信息学分析，寻找转录因子和微小RNA（miRNA）靶位点。结果与对照组相比，有653个基因在骨肉瘤细胞中的表达差异倍数在2倍以上。SELDI—TOFMS显示，在骨肉瘤患者血清中有6个差异表达的质谱峰。通过Link-test分析，共鉴定出13个差异表达的骨肉瘤血清标志蛋白。POLR3F的启动子区存在保守的信号转导和转录激活因子3（STAT3）结合位点，在3’-UTR保守区域含有1个miRNA靶位点。结论Link-test能够有效地从转录组和蛋白质组数据中筛选骨肉瘤标志蛋白。POLR3F有可能成为骨肉瘤早期诊断的标志蛋白和骨肉瘤治疗的新靶点。

Objective To screen potential serum marker proteins of osteosarcoma, and to make a preliminary bioinformatics analysis of RNA polymerase Ⅲ polypeptide F （POLR3F）. Methods Gene chip and SELDI-TOF MS was used to screen genes differentially expressed in osteosarcoma. The associations of potential biomarkers from SELDI data and microarray analysis were further inferred by link-test to identify biomarkers that could likely be used for diagnosis. MATLAB was used to search transcription factors binding site in the promoter region and miRNAs binding site in 3＇-UTR of POLR3F, respectively. Results 653 differentially expressed genes were found in osteosarcoma cells, while six differentially expressed protein peaks with significant statistical significances were detected by SELDI-TOF MS in patient＇s serum. 13 potential biomarkers for early diagnosis of osteosarcoma were screened by link-test. A conserved STAT3 binding site and a miRNA target site were found in proximal promoter regions and 3＇-UTR region of POLR3F, respectively. Conclusions Link-test is a effective method to identify osteosarcoma biomarkers from both microarray and SELDI-TOF MS database. The results confirmed that POLR3F may be a promising biomarker for early diagnosis and a therapy target of osteosarcoma.