ﺍﺭﺯﻳﺎﺑﻲ ﺑﻴﺎﻥ ﮊﻥﻫﺎﻱBax ﻭ Bcl2 ﺩﺭ ﺁﺳﺘﺮﻭﺳﻴﺖﻫﺎﻱ ﺟﺪﺍﺷﺪﻩ ﺍﺯ ﺑﺎﻓﺖ ﻣﻐﺰ ﺍﻧﺴﺎﻥ ﺗﺤﺖ ﺍﺛﺮ ﺍﮔﺰﻭﺯﻭﻡﻫﺎﻱ ﺳﺮﻃﺎﻥ ﺧﻮﻥ

INVESTIGATION OF BAX an‎d BCL2 GENE EXPRESSION IN ASTROCYTES ISOLATED FROM HUMAN BRAIN TISSUE UNDER THE EFFECTS OF LEUKEMIA EXOSOMES

ﺗﺎرﯾﺦ درﯾﺎﻓﺖ 1399/06/07 ﺗﺎرﯾﺦ ﭘﺬﯾﺮش 1399/10/06 ﭼﮑﻴﺪﻩ ﭘﻴﺶﺯﻣﻴﻨﻪ ﻭ ﻫﺪﻑ: ﺗﻬﺎﺟﻢ ﺳﺮﻃﺎﻥﻫﺎ ﺍﺯﺟﻤﻠﻪ ﻟﻮﺳﻤﻲ ﺑﻪ ﺳﻴﺴﺘﻢ ﻋﺼﺒﻲ ﻣﺮﮐﺰﻱ ﺍﺗﻔﺎﻕ ﻣﻲﺍﻓﺘﺪ، ﺳﻠﻮﻝﻫﺎﻱ ﺳﺮﻃﺎﻧﻲ ﻭﻳﮋﮔﻲ ﺗﮑﺜﻴﺮ ﻭ ﻣﻘﺎﻭﻣﺖ ﺩﺭ ﺑﺮﺍﺑﺮ ﺁﭘﻮﭘﺘﻮﺯ ﺩﺍﺭﻧﺪ. ﺍﺯ ﻃﺮﻓﻲ ﺍﮔﺰﻭﺯﻭﻡﻫﺎ ﺑﺎ ﺍﻳﺠﺎﺩ ﺍﺭﺗﺒﺎﻃﺎﺕ ﺳﻠﻮﻝ ﺑﻪ ﺳﻠﻮﻝ ﻣﻮﺟﺐ ﭘﻴﺸﺮﻓﺖ ﺳﺮﻃﺎﻥﻫﺎ ﻣﻲﺷﻮﻧﺪ، ﺩﺭ ﺍﻳﻦ ﻣﻄﺎﻟﻌﻪ ﺍﺛﺮ ﺍﮔﺰﻭﺯﻭﻡﻫﺎﻱ ﺳﺮﻃﺎﻥ ﺧﻮﻥ ﺑﺮ ﺑﻴﺎﻥ ﮊﻥﻫﺎﻱ ﻣﺮﺗﺒﻂ ﺑﺎ ﺁﭘﻮﭘﺘﻮﺯ ﺩﺭ ﺁﺳﺘﺮﻭﺳﻴﺖﻫﺎ ﺑﺮﺭﺳﻲ ﮔﺮﺩﻳﺪ. ﻣﻮﺍﺩ ﻭ ﺭﻭﺵ ﮐﺎﺭ: ﭘﺲ ﺍﺯ ﮐﺸﺖ ﺍﻭﻟﻴﻪ ﺳﻠﻮﻝﻫﺎﻱ ﻋﺼﺒﻲ ﺍﺳﺘﺨﺮﺍﺝﺷﺪﻩ ﺍﺯ ﺑﺎﻓﺖ ﻣﻐﺰ، ﻭ ﺍﺭﺯﻳﺎﺑﻲ ﺍﻳﻤﻮﻧﻮﺳﻴﺘﻮﺷﻴﻤﻲ ﺗﻮﺳﻂ ﺁﻧﺘﻲﺑﺎﺩﻱﻫﺎﻱ SOX2،Nestin ﻭ GFAP، ﺍﺯ ﺭﻭﺵ ﺗﺮﻳﭙﺴﻴﻨﺎﮊ ﺟﻬﺖ ﺟﺪﺍﺳﺎﺯﻱ ﺁﺳﺘﺮﻭﺳﻴﺖﻫﺎ ﺍﺳﺘﻔﺎﺩﻩ ﮔﺮﺩﻳﺪ. ﺍﮔﺰﻭﺯﻭﻡﻫﺎ ﺍﺯ ﻣﺤﻴﻂ ﮐﺸﺖ ﺭﺩﻩﻱ Nalm6 ﺑﻪ ﺭﻭﺵ ﺍﻭﻟﺘﺮﺍﺳﺎﻧﺘﺮﻳﻔﻴﻮﮊ ﺍﺳﺘﺨﺮﺍﺝ ﮔﺮﺩﻳﺪ ﻭ ﺍﺛﺮﺍﺕ ﺁﻥﻫﺎ ﺑﺮ ﺗﮑﺜﻴﺮ ﻭ ﺑﻴﺎﻥ ﮊﻥﻫﺎﻱ Bax ﻭ Bcl2 ﺑﻪ ﺗﺮﺗﻴﺐ ﺗﻮﺳﻂ ﺁﺯﻣﻮﻥ DAPI ﻭ Real time PCR ﺩﺭ ﺁﺳﺘﺮﻭﺳﻴﺖﻫﺎ ﺑﺮﺭﺳﻲ ﮔﺮﺩﻳﺪ. ﻳﺎﻓﺘﻪﻫﺎ: ﻧﺘﺎﻳﺞ ﺍﻳﻤﻮﻧﻮﺳﻴﺘﻮﺷﻴﻤﻲ ﺧﻠﻮﺹ ﺑﺎﻻﻱ 90ﺩﺭﺻﺪ ﺁﺳﺘﺮﻭﺳﻴﺖﻫﺎ ﺭﺍ ﻧﺸﺎﻥ ﺩﺍﺩ. ﺁﺯﻣﻮﻥ DAPI ﭘﺲ ﺍﺯ ﮔﺬﺷﺖ 48 ﺳﺎﻋﺖ ﻧﺸﺎﻥﺩﻫﻨﺪﻩﻱ ﺍﻓﺰﺍﻳﺶ ﻣﻌﻨﺎﺩﺍﺭ ﺁﺳﺘﺮﻭﺳﻴﺖﻫﺎ ﺗﺤﺖ ﺗﻴﻤﺎﺭ ﺑﺎ ﺍﮔﺰﻭﺯﻭﻡ )50µg/ml( ﻧﺴﺒﺖ ﺑﻪ ﮔﺮﻭﻩ ﮐﻨﺘﺮﻝ ﻣﻲﺑﺎﺷﺪ )0/001

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.