The effect of microvesicles derived from K562 cells on proliferation and apoptosis of human bone marrow mesenchymal stem cells

Document Type : Original Article

Authors

1 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University,Tehran, Iran

2 Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

3 Stem Cell and Tissue Engineering Research Center, Shahroud University of Medical Sciences, Shahroud, Iran

4 HSCT Research Center, Laboratory Hematology and Blood Banking Department, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

5 HSCT Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Abstract

Objective(s): Microvesicles (MVs) are small membrane-bound particles that act as a vehicle to transfer their contents, such as proteins, RNAs, and miRNAs, to the target cells, making them undergo several changes. Depending on the origin and the target cell, MVs may cause cell survival or apoptosis. This study investigated the effects of MVs released from the leukemic K562 cell line on the human bone marrow mesenchymal stem cells (hBM-MSCs) to evaluate changes in the survival or apoptosis of the cells in an in vitro system.  
Materials and Methods: In this experimental study, we added the isolated MVs from the K562 cell line to hBM-MSCs, and after three and then seven days, subsequently cell count, cell viability, transmission electron microscopy, tracing MVs by carboxyfluorescein diacetate, succinimidyl ester (CFSE) solution, flow cytometry analysis for Annexin-V/PI staining and qPCR for the evaluation of BCL-2, KI67, and BAX expression were carried out. On the 10th day of the culture, hBM-MSCs were examined by Oil red O and Alizarin Red staining to evaluate their differentiation into adipocytes and osteoblasts.
Results: There was a significant decrease in cell viability and KI67 and BCL-2 expression; however, BAX was significantly upregulated in the hBM-MSCs compared to control groups. Annexin-V/PI staining results also showed the apoptotic effects of K562-MVs on hBM-MSCs. Moreover, the differentiation of hBM-MSCs into adipocytes and osteoblasts was not observed. 
Conclusion: MVs from the leukemic cell line could affect the viability of normal hBM-MSCs and induce cell apoptosis.

Keywords


1. Bettiol A, Marconi E, Lombardi N, Crescioli G, Gherlinzoni F, et al. Pattern of use and long-term safety of tyrosine kinase inhibitors: A decade of real-world management of chronic myeloid leukemia. Clin Drug Investig 2018;38:837-844.
2. Siegel RL, Miller KD, and Jemal A. Cancer statistics, 2016. CA Cancer J Clin 2016; 66:7–30. 
3. Xu Z, Wang H, Wei S, Wang Z, and Ji G. Inhibition of ER stress-related IRE1α/CREB/NLRP1 pathway promotes the apoptosis of human chronic myelogenous leukemia cell. Mol Immunol 2018; 101:377–385. 
4. Luskin MR. Chronic myeloid leukemia and pregnancy: patient and partner perspectives. Expert Rev Hematol 2018; 11:597–599. 
5. Srutova K, Curik N, Burda P, Savvulidi F, Silvestri G, Trotta R, et al. BCR-ABL1 mediated miR-150 downregulation through MYC contributed to myeloid differentiation block and drug resistance in chronic myeloid leukemia. Haematologica 2018; 103:2016–2025. 
6. Zhang L, Chi Y, Wei Y, Zhang W, Wang F, Zhang L, et al. Bone marrow-derived mesenchymal stem/stromal cells in patients with acute myeloid leukemia reveal transcriptome alterations and deficiency in cellular vitality. Stem Cell Res Ther 2021;12:365. 
7. Azadniv M, Myers JR, McMurray HR, Guo N, Rock P, Coppage ML, et al. Bone marrow mesenchymal stromal cells from acute myelogenous leukemia patients demonstrate adipogenic differentiation propensity with implications for leukemia cell support. Leukemia 2020; 34:391-403. 
8. Shen N, Jiang L, Li Q, Cui J, Zhou S, Cheng F, et al. The epigenetic effect of microRNA in BCR-ABL1-positive microvesicles during the transformation of normal hematopoietic transplants. Oncol Rep 2017; 38:3278–3284. 
9. Fonseka P, Marzan AL, and Mathivanan S. Introduction to the Community of Extracellular Vesicles. In: Subcellular Biochemistry. 2021; 3–18. 
10. Abbaszade Dibavar M, Pourbagheri-Sigaroodi A, Asemani Y, Salari S, Bashash D. Extracellular vesicles (EVs): What we know of the mesmerizing roles of these tiny vesicles in hematological malignancies? Life Sciences 271: 2021; 119177. 
11. Akyurekli C, Le Y, Richardson RB, Fergusson D, Tay J, and Allan DS. A Systematic Review of Preclinical Studies on the Therapeutic Potential of Mesenchymal Stromal Cell-Derived Microvesicles. Stem Cell Rev Reports 2015; 11:150–160. 
12. Lopez-Verrilli MA, Caviedes A, Cabrera A, Sandoval S, Wyneken U, and Khoury M. Mesenchymal stem cell-derived exosomes from different sources selectively promote neuritic outgrowth. Neuroscience 2016; 320:129–139. 
13. Zhu X, You Y, Li Q, Zeng C, Fu F, Guo A, et al. BCR-ABL1-positive microvesicles transform normal hematopoietic transplants through genomic instability: Implications for donor cell leukemia. Leukemia 2014; 28:1666–1675. 
14. Hu W, Liu C, Bi ZY, Zhou Q, Zhang H, Li LL, et al. Comprehensive landscape of extracellular vesicle-derived RNAs in cancer initiation, progression, metastasis and cancer immunology. Mol Cancer 2020; 19:102. 
15. Paggetti J, Haderk F, Seiffert M, Janji B, Distler U, Ammerlaan W, et al. Exosomes released by chronic lymphocytic leukemia cells induce the transition of stromal cells into cancer-associated fibroblasts. Blood 2015; 126:1106–1117. 
16. Abbasalipour M, Khosravi MA, Zeinali S, Khanahmad H, Karimipoor M, and Azadmanesh K. Improvement of K562 Cell Line Transduction by FBS Mediated Attachment to the Cell Culture Plate. Biomed Res Int 2019; 2019. 
17. Abbaszade Dibavar M, Soleimani M, Atashi A, Rassaei N, and Amiri S. The effect of simultaneous administration of arsenic trioxide and microvesicles derived from human bone marrow mesenchymal stem cells on cell proliferation and apoptosis of acute myeloid leukemia cell line. Artificial Cells, Nanomedicine, and Biotechnology 2018; 46:S138–S146. 
18. Comparing MTT and SRB.pdf. 
19. Assays CB, Cycle C, Proliferation C, Death C, Assays CB, Cycle C, et al. Detection of Apoptosis Using the BD Annexin V FITC Assay on the BD FACSVerseTM System. BD Biosci 2016; 8:1–2. 
20. Fathi E, Farahzadi R, Valipour B, and Sanaat Z. Cytokines secreted from bone marrow derived mesenchymal stem cells promote apoptosis and change cell cycle distribution of K562 cell line as clinical agent in cell transplantation. PLoS One 2019; 14:1–17. 
21. Zhao P. Exosome derived from bone marrow mesenchymal stem cells improves osteoporosis. 2018; 3962–3970. 
22. Camussi G, Deregibus M-C, Bruno S, Grange C, Fonsato V, and Tetta C. Exosome/microvesicle-mediated epigenetic reprogramming of cells. Am J Cancer Res 2011; 1:98–110. 
23. Jafarzadeh N, Safari Z, Pornour M, Amirizadeh N, Forouzandeh Moghadam M, and Sadeghizadeh M. Alteration of cellular and immune-related properties of bone marrow mesenchymal stem cells and macrophages by K562 chronic myeloid leukemia cell derived exosomes. J Cell Physiol 2019; 234:3697–3710. 
24. Meldolesi J. Exosomes and Ectosomes in Intercellular Communication. Current Biology 2018; 28: R435–R444. 
25. Roccaro AM, Sacco A, Maiso P, Azab AK, Tai YT, Reagan M, et al. BM mesenchymal stromal cell-derived exosomes facilitate multiple myeloma progression. J Clin Invest 2013; 123:1542–1555. 
26. French KC, Antonyak MA, and Cerione RA. Extracellular vesicle docking at the cellular port: Extracellular vesicle binding and uptake. In: Seminars in Cell & Developmental Biology. Elsevier; 2017; 48–55. 
27. Zhao A, Kong F, Liu CJ, Yan G, Gao F, Guo H, et al. Tumor cell-derived microvesicles induced not epithelial-mesenchymal transition but apoptosis in human proximal tubular (HK-2) cells: Implications for renal impairment in multiple myeloma. Int J Mol Sci 2017; 18:1–16. 
28. Ghosh AK, Secreto CR, Knox TR, Ding W, Mukhopadhyay D, and Kay NE. Circulating microvesicles in B-cell chronic lymphocytic leukemia can stimulate marrow stromal cells: Implications for disease progression. Blood 2010; 115:1755–1764. 
29. Penault-Llorca F and Radosevic-Robin N. Ki67 assessment in breast cancer: an update. Pathology 2017; 49:166–171. 
30. Renault TT, Dejean LM, and Manon S. A brewing understanding of the regulation of Bax function by Bcl-xL and Bcl-2. Mech Ageing Dev 2017; 161:201–210. 
31. Kim JA, Shim JS, Lee GY, Yim HW, Kim TM, Kim M, et al. Microenvironmental remodeling as a parameter and prognostic factor of heterogeneous leukemogenesis in acute myelogenous leukemia. Cancer Res 2015; 75:2222–2231. 
32. Maes ME, Schlamp CL, Nickells RW. BAX to basics: How the BCL2 gene family controls the death of retinal ganglion cells. Prog Retin Eye Res 2017; 57:1–25.