Volume 31, Issue 12 (March 2021)
Studies in Medical Sciences 2021, 31(12): 912-920 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Zamani Esmati P, Baharara J, Sahab Negah S, Shahrokh abadi K. INVESTIGATION OF BAX AND BCL2 GENE EXPRESSION IN ASTROCYTES ISOLATED FROM HUMAN BRAIN TISSUE UNDER THE EFFECTS OF LEUKEMIA EXOSOMES. Studies in Medical Sciences 2021; 31 (12) :912-920
URL: http://umj.umsu.ac.ir/article-1-5355-en.html
URL: http://umj.umsu.ac.ir/article-1-5355-en.html
Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran Corresponding Author) , baharara78@gmail.com
Abstract: (2649 Views)
Background & Aims: Unfortunately, today cancers such as leukemia metastasize to the central nervous system (CNS). Cancer cells proliferate and resist apoptosis features. On the other hand, exosomes promote cancer through cell-to-cell communication. This study investigated the effect of leukemia exosomes on the expression of apoptosis-related genes in astrocytes.
Materials & Methods: After the primary culture of brain tissue cells, Immunocytochemical evaluation of cells was performed by specific antibodies Nestin, SOX2, and GFAP. Then, astrocytes were purified from other cells by trypsinage. Exosomes were extracted from supernatant of Nalm6 cell line by ultracentrifugation and their effects on proliferation and expression of Bax and Bcl2 genes were assessed by DAPI and Real-time PCR in astrocytes, respectively.
Results: Results indicated a purity of over 90% of astrocytes in cell culture medium. DAPI test after 48 hours showed a significant increase in astrocytes treated with exosome (50µg/ml) compared to the control group (p < 001). Also, the expression of Bax and Bcl2 genes in exosome-treated astrocytes showed a significant decrease and increase compared to the control group (p <0.01 and p <0.05), respectively.
Conclusion: Leukemia exosomes disrupt the balance of astrocyte proliferation and inhibit apoptosis in them by altering the expression of Bax and Bcl2 genes. Therefore, further studies are recommended to identify exact molecular pathways of cancer.
Materials & Methods: After the primary culture of brain tissue cells, Immunocytochemical evaluation of cells was performed by specific antibodies Nestin, SOX2, and GFAP. Then, astrocytes were purified from other cells by trypsinage. Exosomes were extracted from supernatant of Nalm6 cell line by ultracentrifugation and their effects on proliferation and expression of Bax and Bcl2 genes were assessed by DAPI and Real-time PCR in astrocytes, respectively.
Results: Results indicated a purity of over 90% of astrocytes in cell culture medium. DAPI test after 48 hours showed a significant increase in astrocytes treated with exosome (50µg/ml) compared to the control group (p < 001). Also, the expression of Bax and Bcl2 genes in exosome-treated astrocytes showed a significant decrease and increase compared to the control group (p <0.01 and p <0.05), respectively.
Conclusion: Leukemia exosomes disrupt the balance of astrocyte proliferation and inhibit apoptosis in them by altering the expression of Bax and Bcl2 genes. Therefore, further studies are recommended to identify exact molecular pathways of cancer.
Type of Study: Research |
Subject:
Neuroscience
References
1. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. cell 2011;144(5):646-74. [DOI:10.1016/j.cell.2011.02.013] [PMID]
2. Moore CA, Ibrahim M, Kapila A, Bajaj K. Glioblastoma Multiforme in a Patient with Multiple Myeloma: A Case Report and Literature Review. Perm J 2018;22:17-125 [DOI:10.7812/TPP/17-125]
3. Berghoff AS, Preusser M. The inflammatory microenvironment in brain metastases: potential treatment target? Chin Clin Oncol 2015;4(2):21. [Google Scholar]
4. Weidenfeller C, Svendsen CN, Shusta EV. Differentiating embryonic neural progenitor cells induce blood-brain barrier properties. J Neurochem 2007;101(2):555-65. [DOI:10.1111/j.1471-4159.2006.04394.x] [PMID] [PMCID]
5. Kondegowda NG, Fenutria R, Pollack IR, Orthofer M, Garcia-Ocaña A, Penninger JM, et al. Osteoprotegerin and denosumab stimulate human beta cell proliferation through inhibition of the receptor activator of NF-κB ligand pathway. Cell Metab 2015;22(1):77-85. [DOI:10.1016/j.cmet.2015.05.021] [PMID] [PMCID]
6. Sugimoto MA, Sousa LP, Pinho V, Perretti M, Teixeira MM. Resolution of inflammation: what controls its onset? Front Immunol 2016;7:160. [DOI:10.3389/fimmu.2016.00160] [PMID] [PMCID]
7. Gutzeit C, Nagy N, Gentile M, Lyberg K, Gumz J, Vallhov H, et al. Exosomes derived from Burkitt's lymphoma cell lines induce proliferation, differentiation, and class-switch recombination in B cells. J Immunol 2014;192(12):5852-62. [DOI:10.4049/jimmunol.1302068] [PMID] [PMCID]
8. Peinado H, Alečković M, Lavotshkin S, Matei I, Costa-Silva B, Moreno-Bueno G, et al. Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nat Med 2012;18(6):883-91. [DOI:10.1038/nm.2753] [PMID] [PMCID]
9. Kumar B, Garcia M, Weng L, Jung X, Murakami J, Hu X, et al. Acute myeloid leukemia transforms the bone marrow niche into a leukemia-permissive microenvironment through exosome secretion. Leukemia 2018;32(3): 575-87. [DOI:10.1038/leu.2017.259] [PMID] [PMCID]
10. Hanahan D, Weinberg RA. The hallmarks of cancer. cell 2000;100(1):57-70. [DOI:10.1016/S0092-8674(00)81683-9]
11. Reza Noori Daloii M, Fazilaty Mina Tabrizi H. Cancer metastasis, genetic and microenvironmental factors of distant tissue: a review article. Tehran Univ Med J 2013;70(11) :671-83. [Persian] [Google Scholar]
12. Yang L, Wu X-H, Wang D, Luo Ch-Li, Chen L-X. Bladder cancer cell-derived exosomes inhibit tumor cell apoptosis and induce cell proliferation in vitro. Mol Med Rep 2013;8(4):1272-8. [DOI:10.3892/mmr.2013.1634] [PMID]
13. Choi SS, Lee HJ, Lim I, Satoh J-i, Kim SU. Human astrocytes: secretome profiles of cytokines and chemokines. PloS one 2014;9(4):e92325. [DOI:10.1371/journal.pone.0092325] [PMID] [PMCID]
14. Sokolova V, Ludwig A-K, Hornung S, Rotan O, Horn P A, Epple M, et al. Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. Colloids Surf B Biointerfaces 2011;87(1): 146-50. [DOI:10.1016/j.colsurfb.2011.05.013] [PMID]
15. Othman N, Jamal R, Abu N. Cancer-Derived Exosomes as Effectors of Key Inflammation-Related Players. Front immunol 2019;10:2103. [DOI:10.3389/fimmu.2019.02103] [PMID] [PMCID]
16. Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 2001: 25(4):402-8. [DOI:10.1006/meth.2001.1262] [PMID]
17. Sprowls SA, Arsiwala TA, Bumgarner JR, Shah N, Lateef SS, Kielkowski BN, et al. Improving CNS Delivery to Brain Metastases by Blood-Tumor Barrier Disruption. Trends cancer 2019;5(8):495-505. [DOI:10.1016/j.trecan.2019.06.003] [PMID] [PMCID]
18. Doyle LM, Wang MZ. Overview of extracellular vesicles, their origin, composition, purpose, and methods for exosome isolation and analysis. Cells 2019;8(7):727. [DOI:10.3390/cells8070727] [PMID] [PMCID]
19. Izraeli S, Eckert C. Targeted therapy of CNS leukemia? Blood 2017;130(5):562-3. [DOI:10.1182/blood-2017-06-788430] [PMID]
20. Hosonaga M, Saya H, Arima Y. Molecular and cellular mechanisms underlying brain metastasis of breast cancer. Cancer Metastasis Rev 2020;39(3):711-20. [DOI:10.1007/s10555-020-09881-y] [PMID] [PMCID]
21. Langley RR, Fan D, Guo L, Zhang C, Lin Q, Brantley EC, et al. Generation of an immortalized astrocyte cell line from H-2Kb-tsA58 mice to study the role of astrocytes in brain metastasis. Int J Oncol 2009;35(4):665-72. [DOI:10.3892/ijo_00000378] [PMID] [PMCID]
22. Melo SA, Luecke LB, Kahlert C, Fernandez AF, Gammon ST, Kaye J, et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer. Nature 2015;523(7559):177-82. [DOI:10.1038/nature14581] [PMID] [PMCID]
23. Şovrea AS, Boşca AB. Astrocytes reassessment-an evolving concept part one: embryology, biology, morphology and reactivity. J Mol Psychiatry 2013;1(1):18. [DOI:10.1186/2049-9256-1-18] [PMID] [PMCID]
24. Middeldorp J, Hol E. GFAP in health and disease. Progress in neurobiology. Prog Neurobiol 2011;93(3):421-43. [DOI:10.1016/j.pneurobio.2011.01.005] [PMID]
25. Sharifi H, Shafiee A, Molavi G, Razi E, Mousavi N, Sarvizadeh M, et al. Leukemia‐derived exosomes: Bringing oncogenic signals to blood cells. J cell biochem 2019;120(10):16307-15. [DOI:10.1002/jcb.29018] [PMID]
26. Morad G, Otu H H, Dillon S T, Moses M A. Using proteomics profiling to elucidate the interactions of breast cancer-derived exosomes with the blood-brain barrier. ACS Nano. 2019;13(12): 13853-13865. [DOI:10.1021/acsnano.9b04397] [PMID] [PMCID]
27. Skog J, Wurdinger T, van Rijn S, Meijer D, Gainche L, Sena-Esteves M, et al. Glioblastoma microvesicles transport RNA and protein that promote tumor growth and provide diagnostic biomarkers. Nat Cell Biol 2008;10(12):1470-6. [DOI:10.1038/ncb1800] [PMID] [PMCID]
28. Raimondo S, Saieva L, Corrado C, Fontan, S, Flugy A, Rizzo A, et al. Chronic myeloid leukemia-derived exosomes promote tumor growth through an autocrine mechanism. Cell Commun Signal 2015;13(1): 1-12. [DOI:10.1186/s12964-015-0086-x] [PMID] [PMCID]
29. Matsumoto A, Takahashi Y, Nishikawa M, Sano K, Morishita M, Charoenviriyakul C, et al. Accelerated growth of B16 BL 6 tumor in mice through efficient uptake of their own exosomes by B16 BL 6 cells. Cancer sci 2017; 108(9):1803-10. [DOI:10.1111/cas.13310] [PMID] [PMCID]
30. Plati J, Bucur O, Khosravi-Far R. Apoptotic cell signaling in cancer progression and therapy. Integr Biol 2011;3(4):279-96. [DOI:10.1039/c0ib00144a] [PMID] [PMCID]
31. Adams JM, Cory S. The BCL-2 arbiters of apoptosis and their growing role as cancer targets. Cell Death Differ 2018;25(1):27-36. [DOI:10.1038/cdd.2017.161] [PMID] [PMCID]
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
