Identification of Novel Genes and Candidate Targets in CML Stem Cells

摘要

Chronic myeloid leukemia (CML) is believed to originate from a normal hematopoietc stem cell acquiring the BCR-ABL fusion gene whose protein product has hyperactive tyrosine kinase activity. Though imatinib mesylate(IM) that targets BCR-ABL kinase activity is now widely used, its curative potential as a single agent is not sure, moreover it is unlikely to eliminate the CML stem cells either, which highlights the necessity to elucidate the molecular mechanism operative in CML stem cells. Previously 16 LongSAGE libraries were established to analyze the CML stem cell and their normal counterparts from various sources. There are numerous novel tags which might represent bona fidetranscripts uniquely expressed in these primitive CML SAGE libraries, which provide us an uniquely opportunity to discovery unknown but important transcripts in these cells. We utilized bioinformatics analyses to sort out these novel tags as the candidates to recover the potential bona fide transcripts, and then with PCR and 3'-RACE (Rapid Amplification of cDNA End) approaches we assessed the validity of them and recovered the 3'-end of the potential novel transcripts originated from these tags with the sequence confirmation. The 5'-RACE is under way to eventually recovery the full-length cDNAs and their gene expression pattern between multiple CML and normal primitive cell samples will be assessed.