论文部分内容阅读
本研究采用“cDNA表达文库的血清学分析(serologicalanalysisofcDNAexpressionlibrary,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、cDNAFLJ46859fis、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。
In this study, cDNA library of myeloma HMy2 cells was screened by serologicalanalysis of cDNA library (SEREX). The 30 positive clones obtained were all sequenced, and BLAST homologous sequence alignment analysis. The results showed that there were 6 known genes and 12 new myeloma related tumor antigens. Six ESTs such as loop finger protein 167, KLF10 factor, TPT1 protein, p02 protein, cDNAFLJ46859fis, DNMT1 methyltransferase and so on have been identified by partial sequence ESTs, which are related to the formation, development and prognosis of other tumors. The preliminary analysis and prediction of the new gene structure and function using bioinformatics showed that the complete coding region of MMSA-3, MMSA-8 and MMSA-11 encoded the proteins with length of 215, 160 and 122 amino acids, respectively. Besides MMSA -1 may be located on the sex chromosomes, the rest may be located in the autosomes; MMSA-4 and transcriptional control of tumor proteins are highly similar, MMSA-5 may be involved in phagocytic cells, MMSA-7 may make NF-κB inactivation, MMSA -12 may be lymphocyte cytoplasmic protein. The preliminary analysis of CrELISA showed that the new genes such as MMSA-3 and MMSA-7 were highly specific. Conclusion: The tumor antigens screened and identified in this study can be used in early immunological diagnosis of multiple myeloma, monitoring of residual lesions, prognosis and tumor vaccine preparation.