Establishment and Characterization of a Human Acute Myelocytic Leukemic Cell Line, SH-2, Carrying t(16;17)(q24;q12).

We present a novel human myeloid leukemic cell line, designated as SH-2 which was established from the bone marrow of a patient with acute myelocytic leukemia (AML-M2a) carrying t(16;17)(q24;q12) translocation. The cell line has proliferated continuously in vitro for more than 12 months. Its morphology showed typical features of acute myelocytic leukemia (AML). The cell line’s immunoprofile was accordant with AML (positivity for CD13, CD33, CD38, CD117, CD16, CD56 and MPO). Karyotypic analysis revealed the translocation t(16;17)(q24;q12), monosomy 17 and trisomy 19. The apoptosis related genes such as bcl-2, Fas and GST-πtranscription were detected by RT-PCR. Meanwhile, MDR1, MRP and LRP transcription were not detected by RT-PCR. The deletion of p53 gene and the translocation between chromosomes 16 and 17 were confirmed by FISH method. The SH-2 cells grew colonies in in vitro methylcellulose cultures. Tumor masses were found in 1/2 mice injected by the tail vein with the SH-2 cell line after two months. Infection of the EBV and the mycoplasma were also excluded. Cell line authentication by STR showed that the primary leukemia cell of the patient and the SH-2 cell line originated from same individual. SH-2 cells were proliferated by the addition of cytokines such as IL-3, GM-CSF and SCF. two point mutations in exon 5 of the p53 gene were detected in the SH-2 cells by PCR analysis and direct sequencing showing the conversion of T to G in both codon 349 and 417. The establishment of an myelocytic leukemia cell line with t(16;17)(q24;q12) could be valuable for the study of leukemogenesis and for the research of cloning the new gene involved in the t(16;17)(q24;q12) translocation.