Establishment and characterization of a new human acute myelocytic leukemia cell line SH-2 with a loss of Y chromosome, a derivative chromosome 16 resulting.

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Establishment and characterization of a new human acute myelocytic leukemia cell line SH-2 with a loss of Y chromosome, a derivative chromosome 16 resulting from an unbalanced translocation between chromosomes 16 and 17, monosomy 17, trisomy 19, and p53 alteration Huiying Qiu, Yongquan Xue, Jun Zhang, Jinlan Pan, Haiping Dai, Yafang Wu, Yong Wang, Suning Chen, Depei Wu Experimental Hematology Volume 36, Issue 11, Pages 1487-1495 (November 2008) DOI: 10.1016/j.exphem.2008.06.013 Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 1 Morphology of SH-2 cells on Wright's staining showed uniform round shape, large nuclear with fine granulous chromatin, some azurophilic cytoplasmic granules and unusual nucleoli (original magnification ×1000). Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 2 Immunophenotypic features. (A) SH-2 cell line expressed highly CD13, CD33, CD56, part of cells expressed CD117 and HLA-DR. (B) Cells from nude mice tumor expressed CD45 (98.56%) and the same antigens as SH-2 cell line. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 3 An R-banded karyotype of SH-2 cell line showed 45,X,-Y, der(16)t(16;17)(q24;q12), −17, +19. Arrows indicated the abnormal chromosomes. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 4 CP analysis on a metaphase of SH-2 cells using whole CPs 16 (red) and 17 (green) showed that the intact chromosome 16 (red) and 17 (green) are painted end to end, whereas the der(16) is painted simultaneously by a green fluorescence on the terminal portion of its long arm and by a red one on its remainder. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 5 Interphase fluorescence in situ hybridization on SH-2 cells using a locus-specific probe (green) for p53 gene showed only one green fluorescent signal in each cell indicating the loss of one p53 allele. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 6 Multiplex fluorescence in situ hybridization delineated all chromosomal abnormalities: –Y, der(16)t(16;17), −17, +19 revealed by R-banding karyotypic analysis, however, −1, −4, and −21 are random losses. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 7 The result of short tandem repeat from the patient's BM cells (A) and SH-2 cell line (B) showed the loci in the cell line corresponding with the patient's BM cells except the deletion of Y locus. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 8 Sequence analysis of p53 gene. Arrow indicates the conversion of G to T in exon 5 that changed codon 576 from CAG to CAT. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 9 Reverse transcriptase polymerase chain reaction analysis of the genes expression of SH-2 cell line. (A) The expression of cytokines: Lane 1: marker 100-bp DNA ladder; lane 2: interleukin (IL)-6; lane 3: stem cell factor (SCF); lane 4: IL-11; lane 5: β-actin; lane 6: negative control. (B) Lane 1: marker 100-bp DNA ladder; lane 2: β-actin; lane 3: Bcl-2; lane 4: Bax, lane 5: glutathione S-transferase-π (GST-π); lane 6: Fas; lane 7: p21; lane 8: p27; lane 9: negative control. (C) Lane 1: lung resistance protein; lane 2: multidrug resistance-related protein; lane 3: MDR1; lane 4: marker100-bp DNA ladder; lane 5: β-actin; lane 6: WT1; lane 7: Flt3; lane 8: negative control colony-forming ability Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions

Figure 10 (A) Tumor masses were found in hypodermic tissue of nude mice. (B) Tumor masses in bladder of severe combined immunodeficient mouse. (C) Histopathology examination showed that the tumor masses were composed of leukemia cells and blood vessels. (D) The morphological feature of cells from tumor mass. Experimental Hematology 2008 36, 1487-1495DOI: (10.1016/j.exphem.2008.06.013) Copyright © 2008 ISEH - Society for Hematology and Stem Cells Terms and Conditions