In vivo and in vitro effects of heme oxygenase-1 silencing on the survival of acute myelocytic leukemia-M2 cells

Wei,Sixi; Wang,Yating; Chai,Qixiang; Fang,Qin; Zhang,Yaming; Wang,Jishi

doi:10.3892/etm.2015.2209

In vivo and in vitro effects of heme oxygenase-1 silencing on the survival of acute myelocytic leukemia-M2 cells

Authors:
- Sixi Wei
- Yating Wang
- Qixiang Chai
- Qin Fang
- Yaming Zhang
- Jishi Wang
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Affiliations: Department of Hematology, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China, Department of Pharmacy, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China
Published online on: January 23, 2015 https://doi.org/10.3892/etm.2015.2209
Pages: 931-940

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Abstract

The aim of this study was to investigate the in vivo and in vitro effects of targeted heme oxygenase‑1 (HO‑1) silencing on the proliferation and apoptosis of human acute myelocytic leukemia (AML)-M2 cells. Bone marrow mononuclear cells (BMMNCs) were infected by pRNAi‑siHO‑1‑GFP. The viability of the BMMNCs was determined by cell counting kit‑8 (CCK‑8) assay following daunorubicin (DNR) treatment. The apoptotic rate was detected by flow cytometry. The expression levels of HO‑1 and apoptosis‑related genes were detected by quantitative polymerase chain reaction (qPCR) and western blot analysis. An AML‑M2 xenograft mouse model was established. The tumor formation outcomes and survival were observed. The leukocyte and platelet counts and hemoglobin levels were monitored, and the copy numbers of AML1/ETO fusion gene were detected by qPCR. pRNAi‑siHO‑1‑GFP silenced the expression of HO‑1. DNR inhibited cell viability in a time- and dose‑dependent manner. The survival rate of the cells was significantly reduced by infection with pRNAi‑siHO‑1‑GFP. HO‑1 expression in the BMMNCs infected with pRNAi‑siHO‑1‑GFP was downregulated, whereas caspase‑3, ‑8 and ‑9 expression was upregulated compared with that in control BMMNCs. Kasumi‑1 cells were successfully inoculated into nude mice. The rats inoculated with pRNAi‑siHO‑1‑GFP‑transfected Kasumi‑1 cells succumbed to tumors more slowly and survived longer than those inoculated with untransfected Kasumi‑1 cells. Furthermore, the leukocyte and platelet counts and hemoglobin levels were higher and the copy numbers of AML1/ETO fusion gene were lower in the former group. HO‑1 gene silencing may promote the apoptosis of human M2 leukemic cells by inhibiting a caspase‑dependent apoptotic pathway. Targeted silencing of HO‑1 is able to inhibit the proliferation and infiltration of leukemic cells in nude mice and thus prolong their survival. The findings provide valuable experimental evidence for the molecular targeted therapy of M2 leukemia.

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1	Wang YY, Zhou GB, Yin T, et al: AML1-ETO and C-KIT mutation/overexpression in t(8;21) leukemia: implication in stepwise leukemogenesis and response to Gleevec. Proc Natl Acad Sci USA. 102:1104–1109. 2005. View Article : Google Scholar : PubMed/NCBI
2	Van Besien K: Allogeneic transplantation for AML and MDS: GVL versus GVHD and disease recurrence. Hematology Am Soc Hematol Educ Program. 2013:56–62. 2013. View Article : Google Scholar : PubMed/NCBI
3	Yerlikaya A: Expression of heme oxygenase-1 in response to proteasomal inhibition. Protein Pept Lett. 19:1330–1333. 2012. View Article : Google Scholar : PubMed/NCBI
4	Berberat PO, Dambrauskas Z, Gulbinas A, et al: Inhibition of heme oxygenase-1 increases responsiveness of pancreatic cancer cells to anticancer treatment. Clin Cancer Res. 11:3790–3798. 2005. View Article : Google Scholar : PubMed/NCBI
5	McAllister SC, Hansen SG, Ruhl RA, et al: Kaposi sarcoma-associated herpesvirus (KSHV) induces heme oxygenase-1 expression and activity in KSHV-infected endothelial cells. Blood. 103:3465–3473. 2004. View Article : Google Scholar : PubMed/NCBI
6	Was H, Dulak J and Jozkowicz A: Heme oxygenase-1 in tumor biology and therapy. Curr Drug Targets. 11:1551–1570. 2010. View Article : Google Scholar : PubMed/NCBI
7	Furfaro AL, Piras S, Passalacqua M, et al: HO-1 up-regulation: A key point in high-risk neuroblastoma resistance to bortezomib. Biochim Biophys Acta. 1842:613–622. 2014. View Article : Google Scholar : PubMed/NCBI
8	Castilho Á, Aveleira CA, Leal EC, et al: Heme oxygenase-1 protects retinal endothelial cells against high glucose-and oxidative/nitrosative stress-induced toxicity. PLoS One. 7:e424282012. View Article : Google Scholar
9	Zhang L, Liu YL, Chen GX, et al: Heme oxygenase-1 promotes Caco-2 cell proliferation and migration by targeting CTNND1. Chin Med J (Engl). 26:3057–3063. 2013.
10	Kongpetch S, Kukongviriyapan V, Prawan A, Senggunprai L, Dukongviriyapan U and Buranrat B: Crucial role of heme oxygenase-1 on the sensitivity of cholangiocarcinoma cells to chemotherapeutic agents. PLoS One. 7:e349942012. View Article : Google Scholar : PubMed/NCBI
11	Tibullo D, Barbagallo I, Giallongo C, et al: Nuclear translocation of heme oxygenase-1 confers resistance to imatinib in chronic myeloid leukemia cells. Curr Pharm Des. 19:2765–2770. 2013. View Article : Google Scholar
12	Heasman SA, Zaitseva L, Bowles KM, Rushworth SA and Macewan DJ: Protection of acute myeloid leukaemia cells from apoptosis induced by front-line chemotherapeutics is mediated by haem oxygenase-1. Oncotarget. 2:658–668. 2011.PubMed/NCBI
13	Rushworth SA and MacEwan DJ: HO-1 underlies resistance of AML cells to TNF-induced apoptosis. Blood. 111:3793–3801. 2008. View Article : Google Scholar : PubMed/NCBI
14	Ma D, Fang Q, Li Y, et al: Crucial role of heme oxygenase-1 in the sensitivity of acute myeloid leukemia cell line Kasumi-1 to ursolic acid. Anticancer Drugs. 5:406–414. 2014. View Article : Google Scholar
15	Wang JS, Yang C, Fang Q, et al: K562 cell line resistance to nilotinib induced in vitro and preliminary investigation of its mechanisms. Zhonghua Xue Ye Xue Za Zhi. 33:906–910. 2012.(In Chinese).
16	Chen C, Wang JS, Qin D, Yang Y, Yu YY and Fang Q: The effect of retrovirus-mediated HO-1 gene on chronic myeloid leukemia resistance cell K562/A02 apoptosis induced by nilotinib. Zhonghua Xue Ye Xue Za Zhi. 33:383–387. 2012.(In Chinese). PubMed/NCBI
17	Wang JS, Chai BS, Fang Q, He YY, Chen C and Yang C: Effects of HO-1 gene expression on proliferation of imatinib resistant CMl cells. Zhonghua Xue Ye Xue Za Zhi. 32:388–391. 2011.(In Chinese). PubMed/NCBI
18	Gabert J, Beillard E, van der Velden VH, et al: Standardization and quality control studies of ‘real-time’ quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia - a Europe Against Cancer program. Leukemia. 17:2318–2357. 2003. View Article : Google Scholar : PubMed/NCBI
19	Jozkowicz A, Was H and Dulak J: Heme oxygenase-1 in tumors: is it a false friend? Antioxid Redox Signal. 9:2099–2117. 2007. View Article : Google Scholar : PubMed/NCBI
20	Mayerhofer M, Gleixner KV, Mayerhofer J, et al: Targeting of heat shock protein 32 (Hsp32)/heme oxygenase-1(HO-1) in leukemic cells in chronic myeloid leukemia: a novel approach to overcome resistance against imatinib. Blood. 111:2200–2210. 2008. View Article : Google Scholar
21	Pietsch EC, Chan JY, Torti FM and Torti SV: Nrf2 mediates the induction of ferritin H in response to xenobiotics and cancer chemopreventive dithiolethiones. J Biol Chem. 278:2361–2369. 2003. View Article : Google Scholar
22	Ogborne RM, Rushworth SA and O’Connell MA: α-Lipoic acid-induced heme oxygenase-1 expression is mediated by nuclear factor erythroid 2-related factor 2 and p38 mitogen-activated protein kinase in human monocytic cells. Arterioscler Thromb Vasc Biol. 25:2100–2105. 2005. View Article : Google Scholar : PubMed/NCBI
23	Ogborne RM, Rushworth SA, Charalambos CA and O’Connell MA: Haem oxygenase-1: a target for dietary antioxidants. Biochem Soc Trans. 32:1003–1005. 2004. View Article : Google Scholar : PubMed/NCBI
24	Liu XM, Peyton KJ, Ensenat D, et al: Nitric oxide stimulates heme oxygenase-1 gene transcription via the Nrf2/ARE complex to promote vascular smooth muscle cell survival. Cardiovasc Res. 75:381–389. 2007. View Article : Google Scholar : PubMed/NCBI
25	Trekli MC, Riss G, Goralczyk R and Tyrrell RM: Beta-carotene suppresses UVA-induced HO-1 gene expression in cultured FEK4. Free Radic Biol Med. 34:456–464. 2003. View Article : Google Scholar : PubMed/NCBI
26	Rushworth SA, Bowles KM, Raninga P and MacEwan DJ: NF-kappaB-inhibited acute myeloid leukemia cells are rescued from apoptosis by heme oxygenase-1 induction. Cancer Res. 70:2973–2983. 2010. View Article : Google Scholar : PubMed/NCBI
27	Wu RP, Hayashi T, Cottam HB, et al: Nrf2 responses and the therapeutic selectivity of electrophilic compounds in chronic lymphocytic leukemia. Proc Natl Acad Sci USA. 107:7479–7484. 2010. View Article : Google Scholar : PubMed/NCBI
28	Alam J and Cook JL: How many transcription factors does it take to turn on the heme oxygenase-1 gene? Am J Respir Cell Mol Biol. 36:166–174. 2007. View Article : Google Scholar
29	Tsai JR, Wang HM, Liu PL, et al: High expression of heme oxygenase-1 is associated with tumor invasiveness and poor clinical outcome in non-small cell lung cancer patients. Cell Oncol (Dordr). 35:461–471. 2012. View Article : Google Scholar