Identification And Function Analysis Of LncRNA 630:a Novel Long Non-Coding RNA

Objective:Identification and function analysis of long non-coding RNA 630 in T-cell acute lymphoblastic leukemia cells.Methods:1. The total RNA was extracted from T-cell acute lymphoblastic leukemia cells. Deep sequencing was carried out with high-throughput technology for transcriptome profiling in T-cell acute lymphoblastic leukemia Jurkat cell. New transcripts were discovered by bioinformatics comparative analysis upon the human genome data. By analysising the sequences and structures of new transcripts, some long non-coding RNAs were eventually filtered out.2. A candidate long non-coding RNA 630 was identificated in different tissues and cell lines by RT-PCR 3. Small interfering RNAs were designed against long non-coding RNA 630. Chemically synthesized siRNAs against long non-coding RNA 630 were transfected into Jurkat cells.4. The mRNA level of long non-coding RNA 630 was quantified by real-time quantitative PCR 5. The cellular proliferations were evaluated by using CCK8 method. The flow cytometry analysis was used to assess the cell cycle and apoptosis. 6. The plasmid pEGFP-C1-630 was constructed and transfected into Jurkat cells.7. Testing drug sensitivity of cells by observing the influence of pEGFP-C1-630 on Rh123 accumulation in Jurkat cells. Results:1.630 was a novel long non-coding RNA, which was not expressed in normal human cDNA library and not observed in some cell lines, such as A673, CNE, ECA109, A549, HepG2, LN229 and U87.The mRNA level of long non-coding RNA 630 was significantly increased in Jurkat, Jurkat E6-1 and HL60.Conclusion:Long non-coding RNA 630 is one of the novel transcripts in Jurkat cells, which has been seen in a wide variety of tumor. It may contribute to the process of T-ALL.