OBJECTIVETo investigate the role of transcript factor SCL/TAL-1 gene in the erythroid differentiation through the knockdown of SCL/TAL-1 mRNA by RNA interference.

METHODSThe plasmid of pTRIP-dU3-RNAiTALh-EF1a-GFP with SCL/TAL1 shRNA was transfected into EPO-induced K562 cell line with erythroid differentiation via lentiviral vector system and the expression of SCL/TAL-1 mRNA decreased. The plasmid pTRIP-dU3- RNAiluc-EF1-GFP expressing EGFP gene was as control. The mRNA levels of SCL/TAL-1 and erythroid related RhD, GPA, CD47 in the cell lines were detected by RT-PCR, and erythroid antigen CD71, CD235a were examined by flow cytometry.

RESULTS(1) After 48 h of transfect, more than 95% of K562 cells were GFP positive, indicating the infection rate of the plasmids in the K562 cells more than 95%. (2) The results of RT-PCR showed SCL/TAL-1 mRNA expression in the K562 cell line of knockdown of SCL/TAL-1 was significantly lower than that in the control (P < 0.05). The mRNA levels of CD47 and RhD were also significantly lower, however, GPA decreased slightly in comparison with the control. (3) The expressions of CD71 and CD235a markedly reduced in the K562 cell line of knockdown of SCL/TAL-1 with positive rates as 10.4% and 76.5%, while the positive rates in the control as 94.3% and 83.6%.

CONCLUSIONOur findings suggested that transcription factor SCL/TAL-1 might play an positive role in erythroid differentiation.