多发性骨髓瘤相关抗原免疫学筛选及生物信息学分析.docVIP

多发性骨髓瘤相关抗原免疫学筛选及生物信息学分析 　 作者：周芙玲，张王刚，陈刚，赵万红，曹星梅，陈银霞 田纬，刘苏虎，吴明霞，刘明 【摘要】 本研究采用“cDNA表达文库的血清学分析（serological analysis of cDNA expression library，SEREX）”技术筛选骨髓瘤HMy2 细胞cDNA 表达文库。将得到的30个阳性克隆全部进行测序，并进行BLAST同源序列比对分析。结果表明：获得已知基因6条，骨髓瘤相关肿瘤抗原新基因12条。经部分序列EST拼接，已知基因6条如环指蛋白167、KLF10因子、TPT1蛋白、p02蛋白、cDNA FLJ46859 fis、DNMT1甲基转移酶等，它们与其他肿瘤的形成、发展与预后有一定的关系。利用生物信息学对新基因结构和功能初步分析和预测显示，其中MMSA-3、MMSA-8 和MMSA-11有完整的编码区，编码的蛋白长度分别为215、160和122个氨基酸；除MMSA-1可能定位于性染色体，其余均可能定位于常染色体；MMSA-4和控制转录的肿瘤蛋白高度相似，MMSA-5可能参与细胞的吞噬作用，MMSA-7可能使NF-κB 失活，MMSA-12可能为淋巴细胞的胞质蛋白。CrELISA初步分析表明，MMSA-3 和MMSA-7等新基因特异性较高。结论：本项研究所筛选和鉴定的肿瘤抗原可用于多发性骨髓瘤的早期免疫学诊断、残留病灶监测、判断预后及肿瘤疫苗制备等多种研究。 【关键词】 多发性骨髓瘤 　　Immunological Screening for Multiple Myeloma-Associated Antigens and Their Bioinformatics Analysis 　　　Abstract This study was aimed to screen the cell cDNA expression library of multiple myeloma HMy2 (MM HMy2) by using “serological analysis of cDNA expression library (SEREX)” technique.The obtained 30 positive clones were all sequenced，and analyzed by BLAST (basic local alignment search tool).The results indicated that 6 known genes and 12 new MM-associated genes were obtained，part of which sequences were spliced by EST (expressed sequence tag) splicing.6 known genes such as for ring finger protein 167，KLF10，TPT1 protein，p02 protein，cDNA FLJ46859 fis，DNMT1 methyltrasferase etc.have been demonstrated a certain relationship with other tumors formation，progress and prognosis.The structures and functions of the new genes preliminarily analyzed and predicted by means of bioinformatics showed that MMSA-3，MMSA-8 and MMSA-11 encoding 215，160 and 122 amino acid residues respectively had the full open reading frames (ORF).All the new genes might be located at euchromosomes but MMSA-1 at sex chromosome.MMSA-4 was highly similar to the protein controlling the transcription of tumor antigen，MMSA-5 might take part in cell phagocytosis，MMSA-7 might inactivated NF-κB，and MMSA-12 might be a lymphocytic cytoplasmic protein.The specificity of new genes s