Effects of human bone marrow stromal cell line (HFCL) on the proliferation, differentiation and apoptosis of acute myeloid leukemia cell lines U937, HL-60 and HL-60/VCR
First Online: 21 February 2008 Received: 26 June 2007 Accepted: 05 November 2007 DOI:
Cite this article as: Liang, R., Huang, G., Wang, Z. et al. Int J Hematol (2008) 87: 152. doi:10.1007/s12185-008-0030-6 Abstract
This study investigated the effects of human bone marrow fibroblastoid stromal cell line (HFCL) on the proliferation, differentiation and chemosensitivity of acute myeloid leukemia cells (AML) in vitro coculture. By setting up coculture system of sensitive U937, HL-60 cell line and multidrug-resistant (MDR) HL-60/VCR cells in direct contact with human bone marrow fibroblastoid stromal cell line HFCL, or separated by transwell, the proliferation of AML cells cocultured with HFCL cells was inhibited, compared with AML cells alone. And NBT positive cells increased slightly. The percentage of G1 phase cells of AML cells cocultured with HFCL cells was higher than that without HFCL cells, and that of S phase cells was lower. The expression of CD11b and CD14 increased. Meanwhile HL-60 and HL-60/VCR cells treated by TPT were observed to have apoptosis characteristic morphological changes. The proportion of G0/G1 HL-60 and HL-60/VCR cells treated with TPT increased and the sub-G1 increased. The percentage of Annexin V-positive cells and apoptotic cells increased with expression of activated Caspase-3 and the reduced expression of Bcl-2. But when they were cocultured with HFCL cells, the percentage of Annexin V-positive cells and apoptotic cells decreased and sub-G1 reduced. After indirect contact with HFCL cells the expression of activated Caspase-3 decreased and the expression of Bcl-2 increased. After direct contact with HFCL cells for 96 h, the expression levels of 582 genes in HL-60 cells were up-regulated, and 1,323 genes were down-regulated at least twofold by Affymetrix GeneChip Human Genome U133 set A. The expression change in some genes, such as HL14, was confirmed by RT-PCR and northern blot. In a word, HFCL cells could inhibit the proliferation, induce the monocytic differentiation of U937, HL-60 and HL-60/VCR cells, and prevent TPT-induced apoptosis in HL-60 and HL-60/VCR cells via modulation of Bcl-2 and active Caspase-3. Many genes might take part in the influence of HFCL cells on AML cells, which may give important insights into the interaction of bone marrow stromal cells and leukemic cells.
Keywords Bone marrow stromal cells Acute myeloid leukemia Proliferation Differentiation Apoptosis Gene chip References
Jansen M, Yang FC, Cancelas JA, et al. Rac2-deficient hematopoietic stem cells show defective interaction with the hematopoietic microenvironment and long-term engraftment failure. Stem Cells. 2005;23(3):335–46.
Schmidmaier R, Baumann P, Emmerich B, Meinhardt G. Evaluation of chemosensitivity of human bone marrow stromal cells(—differences between common chemotherapeutic drugs. Anticancer Res. 2006;26(1A):347–50.
Mineo I, Norihiro A, Lynn G, Christoph K, Beverly T-S. Human marrow stromal cells activate monocytes to secrete osteopontin, which down-regulates Notch1 gene expression in CD34+ cells. Blood. 2004;103(12):4496–502.
Kim DH, Lee NY, Baek JH, Kim JG, Sohn SK, Suh JS, Lee KS, Lee KB. Prognostic scoring model based on multi-drug resistance status and cytogenetics in adult patients with acute myeloid leukemia. Leuk Lymphoma. 2006;47(3):461–72.
Schlozel C, Lowenverg B. Stimulation of proliferation and differentiation of acute myeloid leukemia cells on a bone marrow stroma in culture. Exp Hematol. 1985;13:664–9.
Takahashi A, Yamamoto K, Okuma M, Okuma M. Bone marrow stromal cells attenuate mitoxantrone cytotoxicity against HL-60 leukemic cells. Anticancer Drugs. 1992;3:647–50.
Garrido SM, Appelbaum FR, Willman CL, Banker DE. Acute myeloid leukemia cells are protected from spontaneous and drug-induced apoptosis by direct contact with a human bone marrow stromal cell line (HS-5). Exp Hematol. 2001;29:448–57.
Bendall LJ, Art D, Kortlepel K, Gottlieb DJ. Bone marrow adherent layers inhibit apoptosis of acute myeloid leukemia cells. Exp Hematol. 1994;22:1252–7.
Konopleva M, Konoplev S, Hu W, Zaritskey AY, Afanasiev BV, Andreeff M. Stromal cells prevent apoptosis of AML cells by up-regulatin of anti-apoptotic proteins. Leukemia. 2002;16:1713–24.
Fortney JE, Zhao W, Wenger SL, Gibson LF. Bone marrow stromal cells regulate caspase-3 activity in leukemic cells during chemotherapy. Leuk Res. 2001;25:901–7.
Jiang XY, Su YL, Yang SG, Song ZX. Study on establishment, biological character and function of human bone marrow stromal cells. J Chinese Hematol. 1988;9:661–4.
Seki T, Tsuji K, Hayato Y, Moritomo T, Ariga T. Garlic and onion oils inhibit proliferation and induce differentiation of HL-60 cells. Cancer Lett. 2000;160:29–35.
Koistinen P, Siitonen T, Mäntymaa P, Savolainen ER. p53 and redox state in etoposide-induced acute myeloblastic leukemia cell death. Leuk Res. 2001;25:1099–105.
Dassonneville L, Wattez N, Baldeyrou B, Mahieu C, Lansiaux A, Banaigs B, Bonnard I, Bailly C. Inhibition of topoisomerase II by the marine alkaloid ascididemin and induction of apoptosis in leukemia cells. Biochem Pharmacol. 2000;60:527–37.
Lopes EC, Garcia MG, Vellon L, Alvarez E, Hajos SE. Correlation between decreased apoptosis and multidrug resistance (MDR) in murine leukemic T cell lines. Leuk Lymphoma. 2001;42:775–87.
Dai C, Krantz SB. Interferon gamma induces upregulation and activation of caspases 1, 3 and 8 to produce apoptosis in human erythroid progenitor cells. Blood. 1999;93:3309–16.
Mudry RE, Fortney JE, York T, Hall BM. Stromal cells regulate survival of B-lineage leukemic cells during chemotherapy. Blood. 2000;96:1926.
Gibson LF. Survival of B lingeage leukemic cell: signals from the bone marrow microenvironment. Leuk Lymphoma. 2002;43:19–27.
Konopleva M, Konoplev S, Hu W, Zaritskey AY, Afanasiev BV, Andreeff M. Stromal cells prevent apoptosis of AML cells by up-regulation of anti-apoptotic proteins. Leukemia. 2002;16:1713–24.
Van der Velde-Zimmerman D, Smits AJ, Verdaasdonk MA, Rademakers LH, Werner N, Spierings DC, De Weger RA, Van den Tweel JG, Joling P. Beta1-integrins dominate cell tranffic of leukemic cells in human bone-marrow stroma. Int J Cancer. 1996;66:225–33.
Liu TX, Zhang JT, Zhang RB. Gene expression networks underlying retinoic acid-induced differentiation of acute promyelocytic leukemia cells. Blood. 2000;96:1496–503.
Zheng L, Wang L, Yamaga S, Bonds W, Beazer-Barclay Y, Kluger Y, Gerstein M, Newburger PE, Berliner N, Weissman SM. Genomic and proteomic analysis of the myeloid differentiation program. Blood. 2001;98:513–23.
Lee KH, Chang MY, Ahn JI, Yu DH, Jung SS, Choi JH, Noh YH, Lee YS, Ahn MJ. Differential gene expression in retinoic acid-induced differentiation of acute promyelocytic leukemia cells, NB4 and HL-60 cells. Biochem Biophys Res Commun. 2002;296:1125–33.
Gitt MA, Barondes SH. Evidence that a human soluble beta-galactoside-binding lectin is encoded by a family of genes. Proc Natl Acad Sci. 1986;83:7603–7.
Baldini A, Gress T, Patel K, Muresu R, Chiariotti L, Williamson P, Boyd Y, Casciano I, Wells V, Bruni CB. Mapping on human and mouse chromosomes of the gene for the beta-galactoside-binding protein, an autocrine-negative growth factor. Genomics. 1993;15:216–8.
Katagiri K, Yokoyama KK, Yamamoto T, Omura S, Irie S, Katagiri T. Lyn and Fgr protein–tyrosine kinases prevent apoptosis during retinoic acid-induced granulocytic differentiation of HL-60 cells. J Biol Chem. 1996;271:1155–7.
Inohara N, Koseki T, Hu Y, Chen S, Nunez G. CLARP, a death effector domain-containing protein interacts with caspase-8 and regulates apoptosis. Proc Natl Acad Sci. 1997;94:10717–22.
Suzuki A, Yamanaka T, Hirose T, Manabe N, Mizuno K, Shimizu M, Akimoto K, Izumi Y, Ohnishi T, Ohno S. Atypical protein kinase C is involved in the evolutionarily conserved par protein complex and plays a critical role in establishing epithelia-specific junctional structures. J Cell Biol. 2001;152:1183–96.
Amieva MR, Vogelmann R, Covacci A, Tompkins LS, Nelson WJ, Falkow S. Disruption of the epithelial apical–junctional complex by
CogA. Science. 2003;300:1430–4.
Paraguassu-Braga FH, Borojevic R, Bouzas LF, Barcinski MA, Bonomo A. Bone marrow stroma inhibits proliferation and apoptosis in leukemic cells through gap junction-mediated cell communication. Cell Death Differ. 2003;10(9):1101–8.
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