Evaluation of Morpholino Antisense Oligos
’
Role on
BCR-ABL
Gene Silencing in the K562 Cell Line
Mahmoud Haririan, M.Sc.1, Saeid Kaviani, Ph.D.1*, Masoud Soleimani, Ph.D.1, Mehrdad Noruzinia, Ph.D.1, Saeid Abroun, Ph.D.1, Soraya Rasi Ghaemi, M.Sc.2,
Bahman Delalat, M.Sc.1
1. Hematology Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2. Anatomical Sciences Department, Faculty of Medical Sciences, Tarbiat Modares University,
Tehran, Iran
* Corresponding Address: P.O.Box: 14155-111, Hematology Department, Tarbiat Modares University, Tehran, Iran
Email: kavianis@modares.ac.ir Received: 29/Aug/2009, Accepted: 30/Nov/2009
Abstract
Objective: Chronic myeloid leukemia (CML) develops when a hematopoietic stem cell
acquires the BCR/ABL fusion gene. This causes these transformed hematopoietic cells
to have a greater than normal proliferation rate. Scientists attempt to improve the CML treatment process by silencing the BCR/ABL oncogene. In this work, we used morpholino
antisense oligos to silence the BCR/ABL oncogene.
Materials and Methods: In this study, the K562 was used as a BCR/ABL fusion-gene
positive cell line and the Jurkat cell line as a control. We explored the inhibiting capac-ity of morpholino antisense oligos in the the expression of the BCR/ABL oncogene and
studied their p210 BCR/ABL suppression, inhibition of cell proliferation and stimulation of apoptosis in the K562 cells after 24 and 48 hours. Endo-Porter was used for delivery of morpholino antisense oligos into cell cytosols. Meanwhile, flow cytometric analysis was performed in order to determine the appropriate concentration of morpholino antisense oligos.
Results: Prolonged exposure of the K562 cell line to the morpholino antisense oligos
targeted against the BCR-ABL gene showed proliferation inhibition as its main feature.
After western blotting, we found that complete silencing of BCR/ABL was achieved, but
flow cytometric analysis showed no broad apoptosis.
Conclusion: The results indicate that the Morpholino antisense oligo is able to inhibit
p210 BCR/ABL; however, it cannot induce broad apoptosis due to co-silencing of BCR.
Keywords: Chronic Myeloid Leukemia, Morpholino Antisense, K562, BCR/ABL Gene
Original Article
ﻪﻣﺪﻘﻣ
ﻪﻛﺪﺷﺎﺑﻰﻣﺎﻫﻲﻤﺳﻮﻟﺯﺍﻞﻜﺷﻦﻴﺘﺴﺨﻧﻦﻣﺰﻣﻱﺪﻴﻳﻮﻠﻴﻣ ﻲﻤﺳﻮﻟ ﻡﺎﻏﺩﺍﻥژﻞﻴﻟﺩﻪﺑﺮﺘﺸﻴﺑﻯﺭﺎﻤﻴﺑﻦﻳﺍﻞﻣﺎﻋ .(1-3) ﺖﺳﺍﻩﺪﺷﻲﻳﺎﺳﺎﻨﺷ
ﻪﺑﻭﻩﺪﺷﺩﺎﺠﻳﺍﺯﺎﺳﻥﻮﺧﻯﺩﺎﻴﻨﺑﻯﺎﻫﻝﻮﻠﺳﺭﺩﻪﻛﺖﺳﺍBCR-ABLﻪﺘﻓﺎﻳ
ﺎﻴﻔﻟﺩﻼﻴﻓﻡﻭﺯﻮﻣﻭﺮﻛ .(4 ،2) ﺩﻮﺷﻰﻣﻪﺘﺧﺎﻨﺷﺎﻴﻔﻟﺩﻼﻴﻓﻡﻭﺯﻮﻣﻭﺮﻛﻥﺍﻮﻨﻋ
ﻞﺑﺎﻘﺘﻣﻰﻳﺎﺟﻪﺑﺎﺟﺯﺍﻪﻛﺖﺳﺍﻩﺪﺷﻩﺎﺗﻮﻛ 22 ﻡﻭﺯﻮﻣﻭﺮﻛﻥﺎﻤﻫﻞﺻﺍﺭﺩ
ﺩﻮﺟﻭﻪﺑ (BCRﻞﻣﺎﺷ) 22 ﻭ (ABLﻞﻣﺎﺷ) 9 ﻡﻭﺯﻮﻣﻭﺮﻛﺪﻨﻠﺑﻯﻭﺯﺎﺑ
ﺩﺭﻭﺁﻰﻣﺩﻮﺟﻭﻪﺑﺍﺭBCR-ABLﻪﺘﻓﺎﻳﻡﺎﻏﺩﺍﻥژ،ﻰﻳﺎﺟﻪﺑﺎﺟﻦﻳﺍ .ﺪﻳﺁﻰﻣ
ﻦﻴﺌﺗﻭﺮﭘﻮﻜﻧﺍﻪﺑﻦﻣﺰﻣ ﻱﺪﻴﻳﻮﻠﻴﻣﻲﻤﺳﻮﻟﻪﺑ ﻼﺘﺒﻣﻥﺍﺭﺎﻤﻴﺑﺮﺜﻛﺍ ﺭﺩﻪﻛ ﻝﺎﻤﺘﺣﺍﻭﺩﺯﺍﻰﻜﻳﺖﻟﺎﺣﻦﻳﺍﺭﺩ .(6 ،5) ﺩﻮﺷﻰﻣﻪﻤﺟﺮﺗ p210 BCR-ABL
ﻉﻮﻧﻭﺩ .(7) ﺩﺭﺍﺩﺩﻮﺟﻭBCRﻭABLﻥژﻦﻴﺑb3a2ﻭb2a2 ﻝﺎﺼﺗﺍ
ﻲﺻﺎﺧﻉﺍﻮﻧﺍﺎﺑ،p230ﻭp190ﻰﻨﻌﻳ،BCR-ABLﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﺯﺍﺮﮕﻳﺩ .ﺖﺳﺍﻁﺎﺒﺗﺭﺍﺭﺩﺎﻫﻲﻤﺳﻮﻟﺯﺍ
ﻥﺪﺷ ﺯﻮﻴﻓ ﻯﺎﻫﻥﺎﻜﻣ ﻞﻴﻟﺩ ﻪﺑ ﻩﺪﺷ ﺮﻛﺫ ﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﻮﻜﻧﺍ ﺭﺩﺮﻴﺜﻜﺗﻱﺎﻘﻟﺍ ﺖﻴﻟﺎﻌﻓﻲﺳﺭﺮﺑﺭﺩ .(8) ﺪﻧﻮﺷﻰﻣﺩﺎﺠﻳﺍ ﻰﻧژﺕﻭﺎﻔﺘﻣ
ﻭp190BCR-ABL ،p210BCR-ABLﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﻮﻜﻧﺍﻂﺳﻮﺗﺎﻫﺖﻴﺳﻮﻛﻮﻟ
ﻩﺪﻳﺩﺮﮔﺺﺨﺸﻣﺵﻮﻣﻥﺪﺑﻞﺧﺍﺩﻭﺖﺸﻛﻂﻳﺍﺮﺷﺭﺩ P230BCR-ABL
ﺖﻴﻟﺎﻌﻓ p210BCR-ABL ،ﺖﻴﻟﺎﻌﻓ ﻦﻳﺮﺗﻦﻴﻳﺎﭘ P230BCR-ABL ﻪﻛ ﺖﺳﺍ
ﺖﺒﺴﻧﺍﺭﻱﺯﺎﻨﻴﻛﻦﻳﺯﻭﺮﻴﺗﺖﻴﻟﺎﻌﻓﻦﻳﺮﺗﺪﻳﺪﺷp190BCR-ABLﻭﻂﺳﻮﺘﻣ
،ﺮﺑﺍﺮﺑ 3/7 ﺐﻴﺗﺮﺗﻪﺑ) ﺖﺳﺍﺭﺍﺩﻝﺎﻣﺮﻧABLﻥژﺯﺍﻪﻠﺻﺎﺣﻦﻴﺌﺗﻭﺮﭘﻪﺑ
BCR-ABL
ﻥژ
ﻥﺎﻴﺑ
ﺭﺩ
ﺲﻨﺳ
ﻰﺘﻧﺁ
ﻮﮕﻴﻟﺍ
ﻮﻨﻴﻟﻮﻓﺭﻮﻣ
ﻯﺭﺎﻬﻣ
ﺮﺛﺍ
ﻰﺳﺭﺮﺑ
K562
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﺩ
،1
،
.
Ph.D.Dﻥﻭﺮﺑﺁﺪﻴﻌﺳ، 1
،.Ph.D.Dﺎﻴﻧﻱﺯﻭﺭﻮﻧﺩﺍﺩﺮﻬﻣ، 1
،
.
Ph.D.Dﻰﻧﺎﻤﻴﻠﺳﺩﻮﻌﺴﻣ، *1
،
.
Ph.D.Dﻰﻧﺎﻳﻭﺎﻛﺪﻴﻌﺳ، 1
،
.
M.Sc.Dﻥﺎﻳﺮﻳﺮﺣﺩﻮﻤﺤﻣ ،1
،.M.Sc.Dﺖﻟﻻﺩﻦﻤﻬﺑ، 2
،.M.Sc.Dﻰﻤﺋﺎﻗﻰﺛﺍﺭﺎﻳﺮﺛ
ﻥﺍﺮﻳﺍ،ﻥﺍﺮﻬﺗ،ﻯژﻮﻟﻮﺗﺎﻤﻫﻩﻭﺮﮔ،ﻲﻜﺷﺰﭘﻩﺪﻜﺸﻧﺍﺩ،ﺱﺭﺪﻣﺖﻴﺑﺮﺗﻩﺎﮕﺸﻧﺍﺩ .1
ﻥﺍﺮﻳﺍ،ﻥﺍﺮﻬﺗ،ﺢﻳﺮﺸﺗﻡﻮﻠﻋﻩﻭﺮﮔ،ﻲﻜﺷﺰﭘﻩﺪﻜﺸﻧﺍﺩ،ﺱﺭﺪﻣﺖﻴﺑﺮﺗﻩﺎﮕﺸﻧﺍﺩ .2
ﻱژﻮﻟﻮﺗﺎﻤﻫﻩﻭﺮﮔ،ﻲﻜﺷﺰﭘﻡﻮﻠﻋﻩﺪﻜﺸﻧﺍﺩ،ﺱﺭﺪﻣﺖﻴﺑﺮﺗﻩﺎﮕﺸﻧﺍﺩ،14115-111 :ﻲﺘﺴﭘﻕﻭﺪﻨﺻ،ﻥﺍﺮﻬﺗ،ﻥﺍﺮﻳﺍ :ﻝﻮﺌﺴﻣﻩﺪﻨﺴﻳﻮﻧﺱﺭﺩﺁ *
Email: kavianis@modares.ac.ir:ﻚﻴﻧﻭﺮﺘﻜﻟﺍﺖﺴﭘ
88/9/9 :ﻪﻟﺎﻘﻣﺵﺮﻳﺬﭘ،88/6/7 :ﻪﻟﺎﻘﻣﺖﻓﺎﻳﺭﺩ
ﻩﺪﻴﻜﭼ
BCR-ABLﻥژﻥﺩﺮﻛﺵﻮﻣﺎﺧﺖﻬﺟﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺮﻴﺛﺎﺗﻲﺳﺭﺮﺑ :ﻑﺪﻫ *
ﻪﺑJurkatﻱﺎﻫﻝﻮﻠﺳﻭﺖﺒﺜﻣBCR-ABLﻰﻟﻮﻠﺳﻩﺩﺭﻥﺍﻮﻨﻋﻪﺑK562ﻱﺎﻫﻝﻮﻠﺳﺯﺍﻪﻌﻟﺎﻄﻣﻦﻳﺍﺭﺩ :ﺎﻫﺵﻭﺭﻭ ﺩﺍﻮﻣ * ﻮﻨﻴﻟﻮﻓﺭﻮﻣﻪﻨﻴﻬﺑﻯﺎﻫﺖﻈﻠﻏﺭﻮﻀﺣﺭﺩK562ﻱﺎﻫﻝﻮﻠﺳﻰﻟﺎﻤﺘﺣﺍﺯﻮﺘﭘﻮﭘﺁﻭﻰﻟﻮﻠﺳﺮﻴﺜﻜﺗﻥﺍﺰﻴﻣﺲﭙﺳ .ﺪﺷﻩﺩﺎﻔﺘﺳﺍﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋ
ﻥژﺯﺍﻞﺻﺎﺣﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﺩﻮﺟﻭﻦﻴﻨﭽﻤﻫ .ﺪﺷﻰﺑﺎﻳﺯﺭﺍﺖﻋﺎﺳ 48 ﻭ 24 ﺯﺍﺲﭘBCR-ABLﻥژﻰﺻﺎﺼﺘﺧﺍﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍ
ﻡﺎﻧﻪﺑﻚﻴﻠﻴﻓﻰﻔﻣﺁﺪﻴﺘﭙﭘﺯﺍﻝﻮﻠﺳﻞﺧﺍﺩﻪﺑﻪﻃﻮﺑﺮﻣﺲﻨﺳﻰﺘﻧﺁﻝﺎﻘﺘﻧﺍﺭﻮﻈﻨﻣﻪﺑ .ﺪﺷﻰﺳﺭﺮﺑﺕﻼﺑﻥﺮﺘﺳﻭﻂﺳﻮﺗBCR-ABLﺏﻮﻴﻌﻣ
ﻰﺳﺭﺮﺑﺩﺭﻮﻣﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﻂﺳﻮﺗﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻪﻨﻴﻬﺑﺖﻈﻠﻏﻭﺪﺷﻩﺩﺎﻔﺘﺳﺍﺐﺳﺎﻨﻣﺖﻈﻠﻏﻦﻴﻴﻌﺗﺯﺍﺲﭘﺮﺗﺭﻮﭘﻭﺪﻧﺍ
.ﺖﻓﺮﮔﺭﺍﺮﻗ
ﺭﺍﺩﻰﻨﻌﻣﺭﻮﻃﻪﺑﻰﻟﻮﻠﺳﺮﻴﺜﻜﺗﻪﻛﺩﺍﺩﻥﺎﺸﻧﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺎﺑK562ﻱﺎﻫﻝﻮﻠﺳﺕﺪﻣﻲﻧﻻﻮﻃﻪﻬﺟﺍﻮﻣ :ﺎﻫﻪﺘﻓﺎﻳ *
24 ﺯﺍﺲﭘ،ﻝﺮﺘﻨﻛﺎﺑﻪﺴﻳﺎﻘﻣﺭﺩ BCR-ABLﻦﻴﺌﺗﻭﺮﭘﺪﻧﺎﺑﻪﻛﺪﻳﺩﺮﮔﺺﺨﺸﻣﺕﻼﺑﻥﺮﺘﺳﻭﻡﺎﺠﻧﺍﺯﺍﺲﭘ .(p≤ 0/01) ﺖﻓﺎﻳﺶﻫﺎﻛ .ﺪﻳﺩﺮﮔﻮﺤﻣﺎﺒﻳﺮﻘﺗﺖﻳﺎﻬﻧﺭﺩ ﻭﻩﺪﺷﻒﻴﻌﺿﺖﻋﺎﺳ 48 ﻭ24 ﺕﺪﻣﺭﺩBCRﺪﻧﺎﺑﻭﻩﺪﻳﺩﺮﮔﻮﺤﻣﺖﻋﺎﺳ ﺪﻳﺎﻤﻧﻮﺤﻣﺍﺭﻑﺪﻫﻦﻴﺌﺗﻭﺮﭘﻱﺍﻪﻈﺣﻼﻣﻞﺑﺎﻗﺕﺭﺪﻗﺎﺑﺖﺳﺍﺭﺩﺎﻗﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻪﻛﺩﺍﺩﻥﺎﺸﻧﺞﻳﺎﺘﻧ :ﻱﺮﻴﮔﻪﺠﻴﺘﻧ * .ﺪﺷﺎﺑﻰﻤﻧﻊﻴﺳﻭﺯﻮﺘﭘﻮﭘﺁﻱﺎﻘﻟﺍﻪﺑﺭﺩﺎﻗBCR-ABLﺎﺑﻥﺎﻣﺯﻢﻫﻝﺎﻣﺮﻧBCRﻥژﻥﺪﺷﺵﻮﻣﺎﺧﻞﻴﻟﺩﻪﺑ ﺎﻣﺍ
BCR-ABLﻥژ،K562ﻲﻟﻮﻠﺳﻩﺩﺭ،ﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ،ﻦﻣﺰﻣﻯﺪﻴﻳﻮﻠﻴﻣﻲﻤﺳﻮﻟ : :ﻥﺎﮔژﺍﻭﺪﻴﻠﻛ *
ﺯﺎﻨﻴﻛﻦﻳﺯﻭﺮﻴﺗﻚﻳﻊﻗﺍﻭﺭﺩABLﻦﻴﺌﺗﻭﺮﭘ .(9) (ﺮﺑﺍﺮﺑ 7 ﻭﺮﺑﺍﺮﺑ 5/4
ﺪﺷﺭﻢﻴﻈﻨﺗﻭﻰﺘﻳﺍﺪﻫﻯﺎﻫﻝﺎﻨﮕﻴﺳﺭﺩﻰﻤﻬﻣﺶﻘﻧﻪﻛﺖﺳﺍﺭﻮﺘﭙﺳﺭﺪﻗﺎﻓ ﺕﺎﻋﻼﻃﺍ BCR ﻦﻴﺌﺗﻭﺮﭘ ﺩﺮﻜﻠﻤﻋ ﺩﺭﻮﻣ ﺭﺩ ﻰﻟﻭ .(10) ﺩﺭﺍﺩ ﻰﻟﻮﻠﺳ
BCR-ABL ﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘ ﻰﻳﺍﺯﻥﺎﻃﺮﺳ ﺕﺭﺪﻗ .ﺖﺳﺍ ﺖﺳﺩ ﺭﺩ ﻰﻤﻛ
ﻯﺍﺭﺩﻝﺎﻣﺮﻧABLﻦﻴﺌﺗﻭﺮﭘﻪﻛﺖﺳﺍﻙﺭﺩﻞﺑﺎﻗﺖﻘﻴﻘﺣﻦﻳﺍﺮﺘﺴﺑﺭﺩ ﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﻪﻛﻰﻟﺎﺣ ﺭﺩ ﺖﺳﺍﻩﺪﺷﻢﻴﻈﻨﺗ ﻯﺯﺎﻨﻴﻛ ﻦﻳﺯﻭﺮﻴﺗﺖﻴﻟﺎﻌﻓ ﺯﺍ ﺝﺭﺎﺧﺮﻴﺜﻜﺗﻪﺑ ﺮﺠﻨﻣﻭ ﻩﺩﻮﺑ ﻝﺎﻌﻓﺭﺍﺪﻳﺎﭘ ﺕﺭﻮﺻﻪﺑ BCR-ABL
.(11) ﺩﻮﺷﻰﻣﺯﻮﺘﭘﻮﭘﺁﺶﻫﺎﻛﻭﻝﻮﻠﺳﻝﺮﺘﻨﻛ
ﻰﻳﺍﺯﻯﺭﺎﻤﻴﺑﺭﺩﻰﻤﻬﻣﺶﻘﻧBCR-ABLﻥژﻮﻜﻧﺍ ﻪﻛﻰﻳﺎﺠﻧﺁﺯﺍ
ﻰﻳﻭﺭﺍﺩﻯﺎﻫﻥﺎﻣﺭﺩﻯﺍﺮﺑﻲﺑﻮﺧﻑﺪﻫﺪﻧﺍﻮﺗﻰﻣﻥژﻦﻳﺍ،ﺩﺭﺍﺩﻰﻤﺳﻮﻟ ﻑﺪﻫﺎﺑﺲﻨﺳﻰﺘﻧﺁﻯﺎﻫﺵﻭﺭ،ﺮﻴﺧﺍﻪﻫﺩ ﺪﻨﭼﺭﺩ .ﺪﺷﺎﺑﻰﻧﺎﻃﺮﺳﺪﺿ
ﻯﺭﺎﻴﺴﺑﺮﻈﻧﻦﻣﺰﻣﻱﺪﻴﻳﻮﻠﻴﻣﻲﻤﺳﻮﻟﺭﺩBCR-ABLﻥﺎﻴﺑﻥﺩﺮﻛﺭﺎﻬﻣ
ﻮﮕﻴﻟﺍ ﺲﻨﺳﻲﺘﻧﺁ ﻮﮕﻴﻟﺍ ﻊﻗﺍﻭ ﺭﺩ .ﺖﺳﺍ ﻩﺩﺮﻛ ﺐﻠﺟ ﺍﺭ ﻥﺎﻘﻘﺤﻣ ﺯﺍ
ﻪﻤﺟﺮﺗ ﻥﺪﺷﺩﻭﺪﺴﻣﺚﻋﺎﺑﻪﻛﺖﺳﺍﻲﺻﺎﺧﻲﻟﺍﻮﺗ ﺎﺑﻰﻳﺎﻫﺪﻴﺗﻮﺌﻠﻛﻮﻧ ﻉﻮﻧ،ﺎﻫﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻥﺎﻴﻣﺯﺍ .(12-14) ﺩﻮﺷﻰﻣﺮﻈﻧﺩﺭﻮﻣmRNA ﺮﺗﺮﺑﺕﺎﻴﺻﻮﺼﺧﺯﺍﻰﺧﺮﺑﻦﺘﺷﺍﺩﻞﻴﻟﺩﻪﺑﺎﻫﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﻪﺘﺴﺑﺍﻭﺮﻴﻏﻱﺎﻫﻮﮕﻴﻟﺍﺮﮕﻳﺩﻉﺍﻮﻧﺍﺮﻳﺎﺳﻑﻼﺧﺮﺑﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ .ﺖﺳﺍ
ﺯﺍ ﻯﺮﺘﻤﻛ ﺩﺍﺪﻌﺗ ﻪﺑ ﻝﻮﻠﺳ ﻞﺧﺍﺩ ﺭﺩ ﻦﻳﺍﺮﺑﺎﻨﺑ .ﺖﺳﺍ RNaseH ﻪﺑ ﻪﺘﺴﺑﺍﻭﺮﻴﻏﻱﺎﻫﺲﻨﺳﻲﺘﻧﺁﺮﺘﺸﻴﺑﻥﻮﭼ؛ﺩﻮﺷﻰﻣﻞﺼﺘﻣﻑﺪﻫﻯﺎﻫRNA ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ ﻪﻤﺟﺮﺗ ﻥﺩﻮﻤﻧﺩﻭﺪﺴﻣﻱﺍﺮﺑﻪﻛ - RNaseH ﻪﺑ
ﻑﺪﻫ mRNA5'ﺭﺩ ﻊﻗﺍﻭ ﻞﻤﻜﻣ ﺖﺳﺍﻱﺎﻫﻲﻟﺍﻮﺗﻪﻴﻠﻋ - ﺪﻧﺮﻴﮔﻲﻣ
2-5 ﺩﻭﺪﺣﺭﻮﺑﺰﻣﻪﻴﺣﺎﻧﻦﻳﺍﻪﻛﻩﺪﺷﻩﺩﺯﻦﻴﻤﺨﺗ .ﺪﻧﺩﺮﮔﻲﻣﻲﺣﺍﺮﻃ
،ﻞﺻﺍﻦﻳﺍﺮﺑ .ﺩﺮﻴﮔﻲﻣﺮﺑﺭﺩﺍﺭﻲﻟﻮﻠﺳRNAﻥﺰﺨﻣﻱﺎﻫﻲﻟﺍﻮﺗﺯﺍﺪﺻﺭﺩ ﺭﺎﺑ 50 ﺎﺗ 20 ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺪﻨﻧﺎﻣRNaseHﻪﺑﻪﺘﺴﺑﺍﻭﺮﻴﻏﻱﺎﻫﻮﮕﻴﻟﺍ
.(15) ﺪﻨﻨﻛﻰﻣﻞﻤﻋﺮﺗﻲﺻﺎﺼﺘﺧﺍRNaseHﻪﺑﻪﺘﺴﺑﺍﻭﻉﺍﻮﻧﺍﻪﺑﺖﺒﺴﻧ
ﻱﺎﻫﻲﻟﺍﻮﺗﻪﺑﺮﺛﻮﻣﻝﺎﺼﺗﺍﻲﻳﺎﻧﺍﻮﺗﺪﺟﺍﻭﻱﺎﻫﻮﻨﻴﻟﻮﻓﺭﻮﻣﺮﮕﻳﺩﻑﺮﻃﺯﺍ ﻭ ﻱﺭﺍﺪﻳﺎﭘ ،ﺖﻤﻴﻗ ﻥﺩﻮﺑ ﻦﻴﻳﺎﭘ ،ﺖﺧﺎﺳ ﻲﮔﺩﺎﺳ .ﺪﻨﺷﺎﺑﻰﻣ ﻲﻜﻴﺘﻧژ
،15) ﺖﺳﺍﺎﻫﻮﻨﻴﻟﻮﻓﺭﻮﻣﺯﺍﻩﺩﺎﻔﺘﺳﺍﻯﺎﻫﺖﻳﺰﻣﺮﮕﻳﺩﺯﺍﺏﺁﺭﺩﺖﻴﻟﻼﺣ
ﺩﺭﻮﻣﻱﺩﺪﻌﺘﻣﻱﺎﻫﻱﺭﺎﻤﻴﺑﺩﺭﻮﻣﺭﺩﺎﻫﺪﻴﺗﻮﺌﻠﻛﻮﻧﻮﮕﻴﻟﺍﻦﻳﺍﺪﻨﭼﺮﻫ .(16
ﻡﺎﻏﺩﺍﻦﻴﺌﺗﻭﺮﭘﺩﺭﻮﻣﺭﺩﺎﻬﻧﺁﺮﺛﺍﻥﻮﻨﻛﺎﺗﻲﻟﻭ (16) ﺪﻧﺍﻪﺘﻓﺮﮔﺭﺍﺮﻗﻩﺩﺎﻔﺘﺳﺍ .ﺖﺳﺍﻪﺘﻓﺮﮕﻧﺭﺍﺮﻗﻲﺑﺎﻳﺯﺭﺍﺩﺭﻮﻣBCR-ABLﻩﺪﺷ ﻩﺩﺎﻔﺘﺳﺍﺎﺑBCR-ABLﻩﺪﺷﻡﺎﻏﺩﺍﻥژﻲﺷﻮﻣﺎﺧﺮﺛﺍ،ﻖﻴﻘﺤﺗﻦﻳﺍﺭﺩ
ﻲﻧﺎﻃﺮﺳﻱﺎﻫﻝﻮﻠﺳﺮﺑﻩﺪﻳﺪﭘﻦﻳﺍﺮﺛﺍﻭﻮﻨﻴﻟﻮﻓﺭﻮﻣﻱﺎﻫﺪﻴﺗﻮﺌﻠﻛﻮﻧﻮﮕﻴﻟﺍﺯﺍ
.ﺖﺳﺍﻪﺘﻓﺮﮔﺭﺍﺮﻗﻲﺳﺭﺮﺑﺩﺭﻮﻣﻩﺪﺷﻡﺎﻏﺩﺍﻱﺎﻫﻦﻴﺌﺗﻭﺮﭘﻥﺎﻴﺑﻭﻞﻣﺎﺣ
ﺎﻫ
ﺵﻭﺭ
ﻭ
ﺩﺍﻮﻣ
JurkatﻭK562ﻱﺎﻫﻝﻮﻠﺳﺖﺸﻛ
ﻱﺎﻫﻝﻮﻠﺳﻭﺖﺒﺜﻣBCR-ABLﻰﻟﻮﻠﺳﻩﺩﺭﻥﺍﻮﻨﻋﻪﺑK562ﻱﺎﻫﻝﻮﻠﺳ (FBS)ﺪﺻﺭﺩ10 ﻩﺍﺮﻤﻫﻪﺑRPMIﻂﻴﺤﻣﺭﺩﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋﻪﺑJurkat ﺖﺸﻛﻂﻴﺤﻣﺭﺩﻡﺮﺳﺶﻘﻧ .ﺪﻧﺪﺷﻩﺩﺍﺩﺖﺸﻛFetal Bovine Serum ﻱﺎﻫﺯﺎﺳﺶﻴﭘﻭﺎﻫﻪﻨﻴﻣﺁﺪﻴﺳﺍﻲﺧﺮﺑﻭﺎﻫﻥﻮﻣﺭﻮﻫ،ﺪﺷﺭﻱﺎﻫﺭﻮﺘﻛﺎﻓﻦﻴﻣﺎﺗ ﻪﺟﺭﺩ 37 ﺭﻮﺗﺎﺑﻮﻜﻧﺍﺭﺩﺎﻫﻝﻮﻠﺳ .ﺪﺷﺎﺑﻲﻣﺏﺮﭼﺪﻴﺳﺍﻭﻚﻴﺌﻠﻛﻮﻧﺪﻴﺳﺍ
ﻂﻴﺤﻣﺖﻋﺎﺳ 48 ﺮﻫﻭﺪﻧﺪﺷﻯﺭﺍﺪﻬﮕﻧCo2ﺪﺻﺭﺩ 5 ﻭﺩﺍﺮﮔﻰﺘﻧﺎﺳ
.(1 ﻞﻜﺷ) ﺪﺷﺾﻳﻮﻌﺗﺖﺸﻛ
JurkatﻭK562ﻱﺎﻫﻝﻮﻠﺳ :1 ﻞﻜﺷ
ﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻭﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻰﺣﺍﺮﻃ ﻲﻔﻨﻣﻝﺮﺘﻨﻛﺖﻨﺳﺭﻮﻠﻓ
AMOD ﺭﺍﺰﻓﺍﻡﺮﻧ ﺯﺍ ﺲﻨﺳﻲﺘﻧﺁ ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﻲﺣﺍﺮﻃ ﻱﺍﺮﺑ ﻱﺎﻫﻲﻟﺍﻮﺗﻪﻣﺎﻧﺮﺑﻦﻳﺍ .ﺪﺷﺎﺑﻲﻣﺏﻭﺖﺤﺗﻪﻣﺎﻧﺮﺑﻚﻳﻪﻛﺪﺷﻩﺩﺎﻔﺘﺳﺍ
ﻪﺑﻁﻮﺑﺮﻣﻱﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻭﻩﺩﺍﺩﺭﺍﺮﻗﻲﺑﺎﻳﺯﺭﺍﺩﺭﻮﻣﺍﺭﻱﺪﻴﺗﻮﺌﻠﻛﻮﻧ
.(17) ﺪﻳﺎﻤﻧﻲﻣﺏﺎﺨﺘﻧﺍﺍﺭﻥﺁ
ﺭﺍﺪﻘﻣ ﻪﺑ ﻩﺰﻴﻠﻴﻓﻮﻴﻟ ﺭﺩﻮﭘ ﺕﺭﻮﺻ ﻪﺑ ﺲﻨﺳﻲﺘﻧﺁ ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﻱﺭﺍﺪﻳﺮﺧ (ﺎﻜﻳﺮﻣﺁ ،Gene Tools) ﺖﻛﺮﺷ ﺯﺍ ﺮﺘﻴﻟﺮﺑ ﺮﺘﻣﻮﻧﺎﻧ 300
:ﺖﺳﺍﺮﻳﺯﺡﺮﺷﻪﺑﻥﺁﻲﻟﺍﻮﺗ .ﺪﻳﺩﺮﮔ
5'CGC GAA GCC CAC CGG GTC CAC CAT G3' ﻱﺎﻣﺩﺭﺩﻭﻩﺩﺮﻛﺕﻮﻜﻴﻟﺍ 0/5 ﻑﺭﺪﻨﭘﺍﻝﺎﻳﻭﺭﺩﺮﺘﻴﻟﻭﺮﻜﻴﻣ 10 ﺭﺍﺪﻘﻣﻪﺑﻭ
.ﺪﺷﻱﺭﺍﺪﻬﮕﻧ -80
ﻩﺩﺎﻔﺘﺳﺍﺩﺭﻮﻣﻲﻔﻨﻣ ﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋ ﻪﺑﻝﺮﺘﻨﻛﺖﻨﺳﺭﻮﻠﻓﺲﻨﺳﻲﺘﻧﺁ (FITC)ﺎﺑﻭﻩﺰﻴﻠﻴﻓﻮﻴﻟﺭﺩﻮﭘﺕﺭﻮﺻﻪﺑﺭﻮﺑﺰﻣﺲﻨﺳﻲﺘﻧﺁ .ﺖﻓﺮﮔﺭﺍﺮﻗ
ﻪﺑ ﻥﺁﻲﻟﺍﻮﺗﻭﺩﻮﺑﻩﺪﺷ ﺖﻨﺳﺭﻮﻠﻓFluorescein Isothiocyanate
.ﺖﺷﺍﺪﻧJurkatﻭK562ﻝﻮﻠﺳﺭﺩﻲﻠﻤﻜﻣﻪﻛﺪﻳﺩﺮﮔﻲﺣﺍﺮﻃﻱﺍﻪﻧﻮﮔ
:ﺖﺳﺍﺮﻳﺯﺡﺮﺷﻪﺑﻥﺁﻲﻟﺍﻮﺗ
5'-CCT CTT ACC TCA GTT ACA ATT TAT A 3'
.ﺪﻳﺩﺮﮔﻯﺭﺍﺪﻬﮕﻧﻭﺕﻮﻜﻴﻟﺍﻰﻠﺒﻗﻪﻧﻮﻤﻧﻖﺑﺎﻄﻣﻭ
ﺮﺗﺭﻮﭘﻭﺪﻧﺍﺐﺳﺎﻨﻣﺖﻈﻠﻏﻰﺑﺎﻳﺯﺭﺍ
ﺮﺘﻣﻰﻠﻴﻣﺮﻫﺭﺩﻝﻮﻠﺳ 5000 ﻥﺍﺰﻴﻣﻪﺑﻲﻟﻮﻠﺳﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳﺭﻮﻈﻨﻣﻦﻳﺪﺑ
ﺮﺘﻣﻭﺮﻜﻴﻣ 10 ﻩﺍﺮﻤﻫﻪﺑﺍﺭﻥﺁﺯﺍﺮﺘﻴﻟﻲﻠﻴﻣﻚﻳﺲﭙﺳ،ﻩﺩﻮﻤﻧﻪﻴﻬﺗﺐﻌﻜﻣ
ﻪﻧﺎﺧ 24 ﺖﻴﻠﭘﻪﻧﺎﺧﺮﻫﻪﺑ ﺖﻨﺳﺭﻮﻠﻓﻝﺮﺘﻨﻛ ﺲﻨﺳﻲﺘﻧﺁﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑ
ﺮﺘﻣﻭﺮﻜﻴﻣ 2 ﺯﺍﺮﺗﺭﻮﭘﻭﺪﻧﺍﻝﻮﻠﺤﻣﻝﺎﻳﺮﺳﻯﺎﻫﺖﻈﻠﻏﻩﺎﮕﻧﺁ .ﺪﺷﻪﺘﺨﻳﺭ .ﺪﺷﻪﻓﺎﺿﺍﻰﻟﺍﻮﺘﻣﻪﻧﺎﺧ 7 ﺭﺩﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 8 ﺎﺗﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑ
Phosphate Buffered Saline(PBS)ﺎﺑﺭﺎﺑ 3 ﺯﺍﺖﻋﺎﺳ 48 ﺯﺍﺲﭘ
ﺮﻈﻧﺯﺍﺖﻨﺳﺭﻮﻠﻓﺱﻮﻜﻌﻣپﻮﻜﺳﻭﺮﻜﻴﻣﺎﺑﻭﺪﺷﻩﺩﺍﺩﻮﺷﻭﺖﺴﺷﻞﻳﺮﺘﺳﺍ ﺖﺳﺍﻲﻬﻳﺪﺑ .ﺪﺷﻲﺳﺭﺮﺑﻝﻮﻠﺳﻞﺧﺍﺩﺭﺩﻥﺁﻱﻮﮕﻟﺍﻭﺖﻨﺳﺭﻮﻠﻓﺕﺪﺷ
ﻪﻄﻘﻧﺖﻨﺳﺭﻮﻠﻓﻭﻝﻮﻠﺳﻞﺧﺍﺩﻪﺑﺐﺳﺎﻨﻣﺩﻭﺭﻭﺮﮕﻧﺎﻴﺑﺮﺸﺘﻨﻣﺖﻨﺳﺭﻮﻠﻓ ﻱﻮﮕﻟﺍﻭﺩﺮﻫﻥﺪﻳﺩ .ﺖﺳﺍﻝﻮﻠﺳﻞﺧﺍﺩﻪﺑﺐﺳﺎﻨﻣﺎﻧﺩﻭﺭﻭﺮﮕﻧﺎﻴﺑﻪﻄﻘﻧ
ﺩﻭﺭﻭﺮﮕﻧﺎﻴﺑﻢﻫﺯﺎﺑﻪﻛﺖﺴﻴﻧﻝﻮﻤﻌﻣﺮﻴﻏﻥﺎﻣﺯﻢﻫﺕﺭﻮﺻﻪﺑﺰﻴﻧﻕﻮﻓ ﺎﺑﻪﻧﺎﺧﻪﻤﻫﻲﺳﺭﺮﺑﺯﺍﺲﭘ .(2 ﻞﻜﺷ) ﺪﺷﺎﺑﻲﻣﻝﻮﻠﺳﻞﺧﺍﺩﻪﺑﺐﺳﺎﻨﻣ
ﺮﻫﻱﺍﺯﺍﻪﺑﻝﻮﻣﻭﺮﻜﻴﻣ 7 ﻪﻨﻴﻬﺑﺖﻈﻠﻏﺖﻨﺳﺭﻮﻠﻓﺱﻮﻜﻌﻣپﻮﻜﺳﻭﺮﻜﻴﻣ .ﺪﺷﺏﺎﺨﺘﻧﺍﺖﺸﻛﻂﻴﺤﻣﺮﺘﻴﻟﻲﻠﻴﻣ
ﺕﺭﻮﻨﻳﺍ پﻮﻜﺳﻭﺮﻜﻴﻣ ﺎﺑ ﺮﺗﺭﻮﭘﻭﺪﻧﺍ ﺐﺳﺎﻨﻣ ﺖﻈﻠﻏ ﻰﺑﺎﻳﺯﺭﺍ :2 ﻞﻜﺷ
ﻞﺧﺍﺩ ﺭﺩ ﻥﺁ ﻱﻮﮕﻟﺍ ﻭ ﺖﻨﺳﺭﻮﻠﻓ ﺕﺪﺷ ﺮﻈﻧ ﺯﺍ ×400 ﺖﻨﺳﺭﻮﻠﻓ
ﻱﺭﻮﻴﻟﺩ ﺮﮕﻧﺎﻴﺑ ﺮﺸﺘﻨﻣ ﺖﻨﺳﺭﻮﻠﻓ ﺖﺳﺍ ﻲﻬﻳﺪﺑ .ﺪﺷ ﻲﺳﺭﺮﺑ ﻝﻮﻠﺳ
.ﺖﺳﺍﺏﻮﻠﻄﻣ
ﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﻭﺲﻨﺳﻲﺘﻧﺁﻪﻨﻴﻬﺑﺖﻈﻠﻏﻰﺑﺎﻳﺯﺭﺍ
ﺮﻫ ﻱﺍﺯﺍ ﻪﺑ ﻝﻮﻠﺳ 100000 ﻥﺍﺰﻴﻣ ﻪﺑ ﻲﻟﻮﻠﺳ ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ
ﻝﻮﻣﻭﺮﻜﻴﻣ 7 ﻩﺍﺮﻤﻫﻪﺑﻥﺁﺯﺍﺮﺘﻴﻟﻲﻠﻴﻣﻚﻳﻭﻩﺩﻮﻤﻧﻪﻴﻬﺗﺐﻌﻜﻣﺮﺘﻣﻲﻠﻴﻣ
ﻪﻓﺎﺿﺍﻪﻧﺎﺧ 24 ﺖﻴﻠﭘﺭﺩﺯﺎﻴﻧﺩﺭﻮﻣﻱﺎﻫﻪﻧﺎﺧﻪﺑ ﺮﺗﺭﻮﭘﻭﺪﻧﺍﺮﺘﻴﻟﻲﻠﻴﻣﺮﺑ
ﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 2 ﺯﺍﺲﻨﺳﻲﺘﻧﺁﻱﺎﻫﺖﻈﻠﻏﻝﺎﻳﺮﺳﺲﭙﺳ .ﺪﺷ
ﻩﺩﻭﺰﻓﺍﻒﻳﺩﺭ 3 ﺭﺩﻰﻟﺍﻮﺘﻣﻪﻧﺎﺧﺶﺷﺭﺩﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 7 ﺎﺗ
ﺩﺭﻮﻣ ﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ ﺮﻈﻧ ﺯﺍ ﺖﻋﺎﺳ 48 ﻭ 24 ﺯﺍ ﺲﭘ ﺎﻫﻪﻧﻮﻤﻧ .ﺪﺷ
ﺩﺮﺳ PBS ﺎﺑ ﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ ﻱﺍﺮﺑ ﺍﺭ ﺎﻫﻝﻮﻠﺳ .ﺖﻓﺮﮔ ﺭﺍﺮﻗ ﻲﺳﺭﺮﺑ
ﺮﻓﺎﺑﺎﺑﺩﺪﺠﻣﺭﻮﻃﻪﺑﻲﻳﻭﺭﻝﻮﻠﺤﻣﻦﺘﺨﻳﺭﺭﻭﺩﺯﺍﺲﭘﻭﻩﺩﺍﺩﻮﺷﻭﺖﺴﺷ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 195 ﺲﭙﺳ .ﺪﺷﻩﺩﺭﻭﺁﺭﺩﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳﺖﻟﺎﺣﻪﺑBinding FITCﺎﺑﻪﮔﻭﮋﻧﻮﻛV-ﻦﻴﺴﻜﻧﺁﺮﺘﻴﻟﻭﺮﻜﻴﻣ 5 ﺎﺑﻲﻟﻮﻠﺳﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳﺯﺍ
ﺲﭘ .ﺪﻳﺩﺮﮔﻪﺑﻮﻜﻧﺍﻚﻳﺭﺎﺗﻂﻴﺤﻣﺭﺩﻭﻕﺎﺗﺍﻱﺎﻣﺩﺭﺩﻪﻘﻴﻗﺩ10 ﺕﺪﻣﻪﺑ
ﻪﺑ Bindingﺮﻓﺎﺑﺯﺍﺮﺘﻴﻟﻭﺮﻜﻴﻣ 190 ﺎﺑﺎﻫﻝﻮﻠﺳ،ﺩﺪﺠﻣﻯﻮﺷﻭﺖﺴﺷﺯﺍ
ﻩﺩﺎﻣﺁﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﺖﻬﺟﺍﺭﺪﻳﺍﺩﻭﺪﻳﺍﻡﻮﻳﺪﻴﭘﻭﺮﭘﺮﺘﻴﻟﻭﺮﻜﻴﻣ 10 ﻩﺍﺮﻤﻫ
ﻰﻟﻮﻠﺳﺮﻴﺜﻜﺗﺯﺍﺖﻌﻧﺎﻤﻣﺭﺩﺮﻴﺛﺎﺗﻦﻳﺮﺘﺸﻴﺑﻪﻛﻲﻳﺎﻫﺖﻈﻠﻏﺲﭙﺳ .ﺪﻧﺪﺷ .ﺪﺷﺪﻳﺪﻧﺎﻛﺕﻼﺑﻥﺮﺘﺳﻭﻱﺍﺮﺑ،ﺪﻨﺘﺷﺍﺩﺯﻮﺘﭘﻮﭘﺁﻩﺪﻨﻨﻛﺎﻘﻟﺍﺎﻳ
ﺕﻼﺑﻥﺮﺘﺳﻭ
ﻩﻭﺮﮔﻭﻩﺪﺷﺭﺎﻤﻴﺗﻩﻭﺮﮔﻭﺩﺭﺩ K562ﻱﺎﻫﻝﻮﻠﺳ،ﻪﻌﻟﺎﻄﻣﻦﻳﺍﺭﺩ ﻥﺎﻴﺑﻭp210BCR-ABL،p160BCRﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﻥﺎﻴﺑﺮﻈﻧﺯﺍﻰﻔﻨﻣﻝﺮﺘﻨﻛ
ﻰﺑﺎﻳﺯﺭﺍﺕﻼﺑﻥﺮﺘﺳﻭﻚﻴﻨﻜﺗﺎﺑﻰﻠﺧﺍﺩﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋﻪﺑ p83BCR related protein
ﻭ 5 ﻱﺎﻫﺖﻈﻠﻏﺎﺑﺕﺭﻭﺎﺠﻣﺭﺩ K562ﻱﺎﻫﻝﻮﻠﺳﻩﺪﺷﺭﺎﻤﻴﺗﻩﻭﺮﮔﺭﺩ .ﺪﺷ
ﻝﻮﻣﻭﺮﻜﻴﻣ 7 ﻩﺍﺮﻤﻫﻪﺑﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺮﺘﻴﻟﻲﻠﻴﻣﺮﺑﻝﻮﻣﻭﺮﻜﻴﻣ 2
ﺕﺭﻭﺎﺠﻣﺭﺩﻱﺎﻫﻝﻮﻠﺳﻲﻔﻨﻣﻝﺮﺘﻨﻛﻩﻭﺮﮔﺭﺩﻭﺮﺗﺭﻮﭘﻭﺪﻧﺍﺮﺘﻴﻟﻲﻠﻴﻣﺮﺑ ﺖﻋﺎﺳ 48 ﻭ 24 ﺕﺪﻣﻪﺑﻰﻔﻨﻣﻝﺮﺘﻨﻛﺖﻨﺳﺭﻮﻠﻓﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ
ﺯﺍﻩﻭﺮﮔﻭﺩﺭﺩﻰﻠﺒﻗﻯﺎﻫﻝﻮﻠﺳﺪﻨﻧﺎﻣﺰﻴﻧJurkatﻱﺎﻫﻝﻮﻠﺳ .ﺪﻨﺘﻓﺮﮔﺭﺍﺮﻗ .ﺪﻧﺪﺷﻪﻌﻟﺎﻄﻣ p83BCR related proteinﻦﻴﺌﺗﻭﺮﭘﻥﺎﻴﺑﺮﻈﻧ
ﻯﻮﺷﻭﺖﺴﺷ ﺯﺍ ﺲﭘ ﺎﻫﻝﻮﻠﺳ ،ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻡﺎﺠﻧﺍ ﻯﺍﺮﺑ
890 ﻩﺍﺮﻤﻫ ﻪﺑ] ﺲﻳﺰﻳﻻ ﺮﻓﺎﺑ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 100 ﻂﺳﻮﺗ ،PBS ﺎﺑ
(PMSF) ﻝﻮﻠﺤﻣ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ 10 ﻭ ﺺﻟﺎﺧﻕﻮﻓﺮﻄﻘﻣ ﺏﺁ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ .ﺪﻧﺪﺷﺰﻴﻟ [ﺭﻻﻮﻣﻲﻠﻴﻣﺪﺻ Phenyl Methyl Sulphonyl Fluoride ﺮﻫﻦﻴﺑﻪﻔﻗﻭﻪﻴﻧﺎﺛ 30 ﺎﺑﻭﻪﻘﻴﻗﺩﻚﻳﺭﺎﺑﺮﻫﻭﺭﺎﺑﻪﺳ،ﺦﻳﻱﻭﺭﻞﺻﺎﺣﻝﻮﻠﺤﻣ
ﺭﺩﻭﻪﻘﻴﻗﺩﻩﺩﺕﺪﻣﻪﺑ 14000 rpmﺭﻭﺩﺎﺑﺲﭙﺳ .ﺪﺷﻥﻮﻴﺳﺎﻜﻧﻮﺳ،ﻪﻠﺣﺮﻣ
ﺭﺩﻭﻪﺘﺷﺍﺩﺮﺑﺍﺭﻲﻳﻭﺭﻝﻮﻠﺤﻣ .ﺪﻳﺩﺮﮔژﻮﻴﻔﻳﺮﺘﻧﺎﺳﺩﺍﺮﮔﻲﺘﻧﺎﺳﻪﺟﺭﺩ 4 ﻱﺎﻣﺩ
ﻱﺍﺮﺑ .ﺪﺷﻱﺭﺍﺪﻬﮕﻧﺩﺍﺮﮔﻲﺘﻧﺎﺳﻪﺟﺭﺩ -20 ﺭﺰﻳﺮﻓﺭﺩﻭﻪﺘﺨﻳﺭﻑﺭﺪﻨﭘﺍﻝﺎﻳﻭ
SDS-PAGEﺍﺭﺎﻫﻪﻧﻮﻤﻧﻥﺍﻮﺗﻲﻣﻩﺪﺷﺝﺍﺮﺨﺘﺳﺍﻦﻴﺌﺗﻭﺮﭘﺖﻴﻔﻴﻛﻲﺳﺭﺮﺑ ﻥﺎﻨﻴﻤﻃﺍﺎﻫﺪﻧﺎﺑﺖﻴﻔﻴﻛﺯﺍﺎﺗﺩﻮﻤﻧﮓﻧﺭﻮﻠﺑﻲﺳﺎﻣﻮﻛﮓﻧﺭﺎﺑﺍﺭﻝژﻭﻩﺩﻮﻤﻧ ﻯﺮﻴﮔﻩﺯﺍﺪﻧﺍBradfordﺵﻭﺭﺎﺑﻩﺪﺷﺝﺍﺮﺨﺘﺳﺍﻦﻴﺌﺗﻭﺮﭘﺖﻈﻠﻏ .ﺩﺮﻛﻞﺻﺎﺣ
ﻱﺍﺮﺑ .ﺪﻳﺩﺮﮔﻱﺭﺍﺬﮔﺭﺎﺑﻦﻴﺌﺗﻭﺮﭘﻡﺮﮔﻭﺮﻜﻴﻣ 50 ﺶﻫﻭﮋﭘﻦﻳﺍﺭﺩﻪﻛ .ﺪﺷ 1 ﻪﺑ 3ﻞﻗﺍﺪﺣﻰﻧﺯﻭﺖﺒﺴﻧﺎﺑﻪﻧﻮﻤﻧﺮﻓﺎﺑSDS-PAGEﺭﺩﻪﻧﻮﻤﻧﻱﺯﺎﺳﻩﺩﺎﻣﺁ ﺯﺍﺪﻌﺑSDS-PAGEﺭﺩ .ﺩﻮﺷﻉﺎﺒﺷﺍSDSﺎﺑﺎﻫﻦﻴﺌﺗﻭﺮﭘﺎﺗﻩﺩﻭﺰﻓﺍﻪﻧﻮﻤﻧﻪﺑ ﻱﺎﻣﺩﺭﺩﺎﻬﻧﺁﻁﻮﻠﺨﻣ،ﻯﺭﺍﺬﮔﻪﻧﻮﻤﻧﺯﺍﻞﺒﻗﻭﻦﻴﺌﺗﻭﺮﭘﻪﺑﻪﻧﻮﻤﻧﺮﻓﺎﺑﻥﺩﻭﺰﻓﺍ ﺎﻫﻦﻴﺌﺗﻭﺮﭘ PAG .ﺪﺷﻩﺩﺍﺩﺭﺍﺮﻗﻪﻘﻴﻗﺩ 12 ﺕﺪﻣﻪﺑﺩﺍﺮﮔﻲﺘﻧﺎﺳﻪﺟﺭﺩ 100
ﺵﻭﺭﻦﻳﺍ .ﺩﻮﺑﻰﻛﺪﻧﺍﺕﺍﺮﻴﻴﻐﺗﺎﺑﻰﻠﻣﻻﺵﻭﺭﺮﺑﻰﻨﺘﺒﻣ،ﻪﻌﻟﺎﻄﻣﻦﻳﺍﺭﺩ
،ﺩﻮﺷﻰﻣﺏﻮﺴﺤﻣSDSﺭﻮﻀﺣﺭﺩﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍﺵﻭﺭﻦﻳﺮﺗﻝﻮﻤﻌﻣﻪﻛ ﺯﺍﺶﻴﺑﻰﻟﻮﻜﻟﻮﻣﻥﺯﻭﺎﺑﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﻯﺯﺎﺳﺍﺪﺟﺭﺩﻰﻳﻻﺎﺑﺭﺎﻴﺴﺑﻰﻳﺍﺭﺎﻛ
10 ﻯﻮﺘﺤﻣ ) x5 ﺮﻓﺎﺑﺎﺑ 1 ﻪﺑ 4 ﺍﺭﻦﻴﺌﺗﻭﺮﭘﻪﻧﻮﻤﻧﺲﭙﺳ .ﺩﺭﺍﺩﻥﻮﺘﻟﺍﺩﻮﻠﻴﻛ 15
ﺮﺘﻴﻟﻰﻠﻴﻣ 0/2 ،SDSﻡﺮﮔ 1 ،ﻝﻭﺮﺴﻴﻠﮔﺮﺘﻴﻟﻰﻠﻴﻣ 5 ،Stakingﺮﻓﺎﺑﺮﺘﻴﻟﻰﻠﻴﻣ
ﻝﻮﻧﺎﺗﺍﻮﺘﭘﺎﻛﺮﻣ -2 ﺮﺘﻴﻟﻰﻠﻴﻣ 1 ﻭ (ﻝﻮﻧﺎﺗﺍﺭﺩﺪﺻﺭﺩ 0/5) ﻮﻠﺑﻞﻨﻓﻮﻣﺮﺑﻝﻮﻠﺤﻣ
ﻪﻧﻮﻤﻧ .ﺪﺷﻁﻮﻠﺨﻣ (ﺪﺷﻩﺪﻧﺎﺳﺭﺮﺘﻴﻟﻰﻠﻴﻣ 20 ﻰﻳﺎﻬﻧﻢﺠﺣﻪﺑﺮﻄﻘﻣﺏﺁﺎﺑﻪﻛ
ﻩﺩﺍﺩﺭﺍﺮﻗﻪﺟﺭﺩ 100 ﻱﺎﻣﺩﺎﺑﻲﺗﺭﺍﺮﺣﺮﻜﻴﺷﺭﺩﻪﻘﻴﻗﺩ 12 ﺕﺪﻣﻪﺑﻪﻧﻮﻤﻧﺮﻓﺎﺑﻭ
Stackingﻝژﺎﻫﻚﻫﺎﭼﺭﺩﺖﻗﺩﻪﺑﻪﻧﻮﻤﻧﺮﻫﺯﺍﺮﺘﻴﻟﻭﺮﻜﻴﻣ 30 ﺲﭙﺳ .ﺪﺷ .ﺪﻳﺩﺮﮔﻡﺎﺠﻧﺍﺖﻟﻭ 80 ﺖﺑﺎﺛژﺎﺘﻟﻭﺎﺑﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍﻭﺪﺷﻪﺘﺨﻳﺭ
ﻰﻟﻮﻠﺳﺕﺎﻴﺣﺪﺻﺭﺩﻰﺑﺎﻳﺯﺭﺍ
ﻢﻫﺎﺑﻰﻟﻮﻠﺳﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳﺯﺍﺮﺘﻴﻟﻭﺮﻜﻴﻣ 50 ﺩﻭﺪﺣﺭﻮﻈﻨﻣﻦﻳﺍﻯﺍﺮﺑ
ﺵﺭﺎﻤﺷﻯﺍﺮﺑﺲﭙﺳ .ﺪﺷﻁﻮﻠﺨﻣﺪﺻﺭﺩ0/4 ﻮﻠﺑﻥﺎﭙﻳﺮﺗ ﺎﺑﺩﻮﺧﻢﺠﺣ
ﺩﺍﺪﻌﺗﻯﺎﻨﺒﻣﺮﺑﻰﻟﻮﻠﺳﺕﺎﻴﺣﺪﺻﺭﺩ .ﺪﻳﺩﺮﮔﻩﺩﺎﻔﺘﺳﺍﺭﺎﺑﻮﺌﻧﻡﻻﺯﺍﺎﻫﻝﻮﻠﺳ
ﺖﺳﺍﺮﻛﺫﻪﺑﻡﺯﻻ .ﺪﻳﺩﺮﮔﻪﺒﺳﺎﺤﻣ،ﺎﻫﻝﻮﻠﺳﻞﻛﺩﺍﺪﻌﺗﻪﺑﻩﺪﻧﺯﻯﺎﻫﻝﻮﻠﺳ .ﺪﻧﺪﺷﺵﺭﺎﻤﺷﻪﻧﺎﮔﺍﺪﺟﺕﺭﻮﺻﻪﺑﻩﺩﺮﻣﻭﻩﺪﻧﺯﻯﺎﻫﻝﻮﻠﺳﻪﻛ
ﻯﺭﺎﻣﺁﻰﺑﺎﻳﺯﺭﺍ
ﻒﻠﺘﺨﻣﻯﺎﻫﺖﻈﻠﻏﺭﻮﻀﺣﺭﺩﺎﻫﻝﻮﻠﺳگﺮﻣﺪﺻﺭﺩﻲﺳﺭﺮﺑﺖﻬﺟ ﺲﭘ (ﻩﻭﺮﮔ 7 ﻦﻴﺑﺭﺩ) ﻰﻔﻨﻣﻝﺮﺘﻨﻛﻩﻭﺮﮔﻭﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ
(Kruskal-Wallis Test)ﺲﻴﻟﺍﻭﻝﺎﻜﺳﻭﺮﻛﺖﺴﺗ،ﺖﻋﺎﺳ 48 ﻭ 24 ﺯﺍ
ﻲﺳﺭﺮﺑﻱﺍﺮﺑ(Mann-Whitney Test)ﻲﻨﺘﻳﻭ –ﻦﻣﺖﺴﺗﺯﺍ .ﺪﺷﻡﺎﺠﻧﺍ
ﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 2 ﺎﺑﻩﺪﺷﺭﺎﻤﻴﺗﻩﻭﺮﮔﻭﺩﻦﻴﺑﺭﺩﺎﻫﻝﻮﻠﺳﻥﺪﻧﺎﻣﻩﺪﻧﺯﺪﺻﺭﺩ .ﺪﺷﻩﺩﺎﻔﺘﺳﺍﻰﻔﻨﻣﻝﺮﺘﻨﻛﻩﻭﺮﮔﻭﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺭﻻﻮﻣﻲﻠﻴﻣ
ﺎﻫ
ﻪﺘﻓﺎﻳ
ﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﺯﺍﻞﺻﺎﺣﺞﻳﺎﺘﻧ
،ﺲﻨﺳﻰﺘﻧﺁ ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﻪﻨﻴﻬﺑ ﺖﻈﻠﻏ ﻯﺮﻴﮔﻩﺯﺍﺪﻧﺍ ﺭﻮﻈﻨﻣ ﻪﺑ
(ﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 6 ﻭ 5 ،4 ،3 ،2) ﺲﻨﺳﻰﺘﻧﺁﺏﻭﺎﻨﺘﻣﻯﺎﻫﺖﻈﻠﻏ
ﻰﻳﺎﺗ24 ﺖﻴﻠﭘﺯﺍﻪﻧﺎﺧﺮﻫﺭﺩﺮﺗﺭﻮﭘﻭﺪﻧﺍﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 7 ﻩﺍﺮﻤﻫﻪﺑ
ﺖﻋﺎﺳ 48 ﻭ 24 ﺯﺍﺲﭘ .ﺪﺷﻩﺩﻭﺰﻓﺍK562ﻰﻟﻮﻠﺳﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳﻯﺍﻮﺘﺤﻣ ﺶﻳﺍﺰﻓﺍﺎﺑﺭﺎﻈﺘﻧﺍﻑﻼﺧﺮﺑﻪﻛﺪﻳﺩﺮﮔﺺﺨﺸﻣﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﻡﺎﺠﻧﺍﺎﺑ ﻪﭼﺮﮔﺍ؛ﺩﺍﺪﻧ ﻥﺎﺸﻧﺍﺭﻰﺸﻫﺎﻛﻰﻟﻮﻠﺳﺮﻴﺜﻜﺗﻥﺍﺰﻴﻣ،ﺲﻨﺳﻰﺘﻧﺁ ﺖﻈﻠﻏ ﺮﺗﻦﻴﻳﺎﭘﻯﺎﻫﺖﻈﻠﻏﻰﻟﻭﺪﺷﻰﻟﻮﻠﺳﺮﻴﺜﻜﺗﺶﻫﺎﻛﺚﻋﺎﺑﺎﻫﺖﻈﻠﻏﻰﻣﺎﻤﺗ ﺭﺩ ﻯﺮﺘﺸﻴﺑ ﺶﻫﺎﻛ ﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑ ﺮﺘﻣﻭﺮﻜﻴﻣ 2 ﻩﺩﻭﺪﺤﻣ ﺎﺗ ﺲﻨﺳﻰﺘﻧﺁ 48 ﻭ 24 ﺯﺍﺲﭘﺎﻫﻝﻮﻠﺳﺯﻮﺘﭘﻮﭘﺁﻥﺍﺰﻴﻣﻦﻴﻨﭽﻤﻫ .ﺩﻮﻤﻧﺎﻘﻟﺍﺎﻫﻝﻮﻠﺳﺮﻴﺜﻜﺗ
ﻰﻟﻭﺰﻧﺮﻴﺳﺲﻨﺳﻰﺘﻧﺁﺖﻈﻠﻏﺶﻳﺍﺰﻓﺍﺎﺑ،ﺲﻨﺳﻰﺘﻧﺁﺎﺑﺕﺭﻭﺎﺠﻣﺖﻋﺎﺳ ﺚﻋﺎﺑﺲﻨﺳﻰﺘﻧﺁﻯﺎﻫﺖﻈﻠﻏﻰﻣﺎﻤﺗﺮﮕﻳﺩﺕﺭﺎﺒﻋﻪﺑ .(3 ﻞﻜﺷ) ﺖﺷﺍﺩ 48 ﺲﭘﺯﻮﺘﭘﻮﭘﺁﻥﺍﺰﻴﻣﻦﻳﺍ .ﺪﺷﻰﻔﻨﻣﻝﺮﺘﻨﻛﻩﻭﺮﮔﺎﺑﻪﺴﻳﺎﻘﻣﺭﺩﺯﻮﺘﭙﭘﺁ
ﺯﻮﺘﭘﻮﭘﺁﺪﺻﺭﺩﻰﻔﻨﻣﻝﺮﺘﻨﻛﻩﻭﺮﮔﺭﺩﻪﻛﻰﻟﺎﺣﺭﺩﺖﻓﺎﻳﺶﻳﺍﺰﻓﺍﺖﻋﺎﺳ
ﻯﺎﻫﻝﻮﻠﺳ ﺩﺭﻮﻣﺭﺩ .(1 ﺭﺍﺩﻮﻤﻧ) ﺩﺍﺩ ﻥﺎﺸﻧ ﺶﻫﺎﻛﻰﺘﺣ ﻩﺩﺮﻜﻧﺮﻴﻴﻐﺗ
.(3 Cﻞﻜﺷ) ﺩﺮﻜﻧﺮﻴﻴﻐﺗﺯﻮﺘﭘﻮﭘﺁﻥﺍﺰﻴﻣﻰﻔﻨﻣﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋﻪﺑJurkat
40 35 30 25 20 15 10 5 0 ﺪﺻ ﺭﺩ
ﺖﻋﺎﺳ 24
ﺖﻋﺎﺳ 48
ﻲﻔﻨﻣ ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑﺭﻻ ﻮﻣﻭﺮﻜﻴﻣ 2 ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑﺭﻻ ﻮﻣﻭﺮﻜﻴﻣ 3 ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑﺭﻻ ﻮﻣﻭﺮﻜﻴﻣ 4 ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑﺭﻻ ﻮﻣﻭﺮﻜﻴﻣ 5 ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑﺭﻻ ﻮﻣﻭﺮﻜﻴﻣ 6 ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑﺭﻻ ﻮﻣﻭﺮﻜﻴﻣ 7
(ﻰﻟﻮﻃﺭﻮﺤﻣ) ﻲﻟﻮﻠﺳگﺮﻣﺪﺻﺭﺩﺐﺴﺣﺮﺑﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﺞﻳﺎﺘﻧ :1ﺭﺍﺩﻮﻤﻧ
ﻮﮕﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻯﺎﻫﺖﻈﻠﻏﻰﻣﺎﻤﺗ .(ﻰﺿﺮﻋﺭﻮﺤﻣ) ﺲﻨﺳﻲﺘﻧﺁﺖﻈﻠﻏﻭ
(ﺮﺘﻴﻟﻲﻠﻴﻣﺮﺑﺭﻻﻮﻣﻭﺮﻜﻴﻣ 7 ﺎﺗﺮﺘﻴﻟﻲﻠﻴﻣﺮﺑﺭﻻﻮﻣﻭﺮﻜﻴﻣ 2 ﺯﺍ) ﺲﻨﺳﻰﺘﻧﺁ
ﻝﺮﺘﻨﻛﻩﻭﺮﮔﺎﺑﻪﺴﻳﺎﻘﻣﺭﺩﺯﻮﺘﭘﻮﭘﺁﻥﺍﺰﻴﻣﻦﻳﺍﻭﺪﻧﺪﺷﺯﻮﺘﭘﻮﭘﺁﺚﻋﺎﺑ
.ﺩﺍﺪﻧﻥﺎﺸﻧﻯﺭﺍﺩﻰﻨﻌﻣﻑﻼﺘﺧﺍﻯﺭﺎﻣﺁﺮﻈﻧﺯﺍﻰﻟﻭﺖﻓﺎﻳﺶﻳﺍﺰﻓﺍﻰﻔﻨﻣ
bcr-ablﻦﻴﺌﺗﻭﺮﭘﻯﻭﺭﺮﺑﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺮﻴﺛﺎﺗ
ﺮﺘﻣﻭﺮﻜﻴﻣ 5 ﺎﺑﺕﺭﻭﺎﺠﻣﺭﺩﻩﺪﺷﺭﺎﻤﻴﺗﻩﻭﺮﮔﺭﺩ K562ﻱﺎﻫﻝﻮﻠﺳ ﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 7 ﻩﺍﺮﻤﻫﻪﺑﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ 2 ﻭﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑ
ﺎﻫﻝﻮﻠﺳ ،ﻲﻔﻨﻣﻝﺮﺘﻨﻛ ﻩﻭﺮﮔﺭﺩ ﻭ ﻪﺘﻓﺮﮔ ﺭﺍﺮﻗ ﺮﺗﺭﻮﭘﻭﺪﻧﺍ ﺭﻻﻮﻣﻲﻠﻴﻣ ﺲﻨﺳﻲﺘﻧﺁ ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﺭﻻﻮﻣﻲﻠﻴﻣ ﺮﺑ ﺮﺘﻣﻭﺮﻜﻴﻣ 2 ﺕﺭﻭﺎﺠﻣ ﺭﺩ
ﺮﻈﻧﺯﺍﻭﺪﻨﺘﻓﺮﮔﺭﺍﺮﻗﺖﻋﺎﺳ 48 ﻭ 24 ﺕﺪﻣﻪﺑﻰﻔﻨﻣﻝﺮﺘﻨﻛﺖﻨﺳﺭﻮﻠﻓ
ﻪﺑ) p83BCR related protein ﻭ p210BCR-ABL ،p160bcrﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘ ﻥﺎﻴﺑ
ﻱﺎﻫﻝﻮﻠﺳ .ﺪﻧﺪﺷﻰﺑﺎﻳﺯﺭﺍﺕﻼﺑﻥﺮﺘﺳﻭﻚﻴﻨﻜﺗﺎﺑ (ﻰﻠﺧﺍﺩﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋ
ﻦﻴﺌﺗﻭﺮﭘﻥﺎﻴﺑﺮﻈﻧﺯﺍﻩﻭﺮﮔﻭﺩﺭﺩﻰﻠﺒﻗﻯﺎﻫﻝﻮﻠﺳﻥﻮﭼﻢﻫﺰﻴﻧJurkat
ﻡﺎﺠﻧﺍ ،K562 ﻯﺎﻫﻝﻮﻠﺳ ﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘ ﻥﺩﺮﻛ ﺝﺍﺮﺨﺘﺳﺍ ﺎﺑ ،ﻯﺪﻴﻳﻮﻠﻛG-250 ﻯﻮﻠﺑﻰﺳﺎﻣﻮﻛﺎﺑﻝژﻯﺰﻴﻣﺁﮓﻧﺭﻭSDS-PAGE ﺎﺑﻩﺪﺷﺝﺍﺮﺨﺘﺳﺍﻦﻴﺌﺗﻭﺮﭘﺐﺳﺎﻨﻣﺖﻴﻔﻴﻛﺯﺍﻰﻛﺎﺣﻩﺪﻣﺁﺖﺳﺩﻪﺑﺞﻳﺎﺘﻧ
.(4 ﻞﻜﺷ) ﺪﺷﺎﺑﻰﻣﻪﻃﻮﺑﺮﻣﻞﻜﺗﻭﺮﭘﺯﺍﻩﺩﺎﻔﺘﺳﺍ
k562ﻱﺎﻫﻝﻮﻠﺳﺯﺍﻩﺪﺷﺝﺍﺮﺨﺘﺳﺍﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘSDS-PAGE :4 ﻞﻜﺷ
.ﺖﺳﺍﺎﻫﻦﻴﺌﺗﻭﺮﭘﺐﺳﺎﻨﻣﺖﻴﻔﻴﻛﻩﺪﻨﻫﺩﻥﺎﺸﻧ
ﻦﻴﺌﺗﻭﺮﭘ ﺪﻧﺎﺑ ﻪﻛ ﺪﻳﺩﺮﮔ ﺺﺨﺸﻣ ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻡﺎﺠﻧﺍ ﺯﺍ ﺲﭘ ﻲﻔﻨﻣﻝﺮﺘﻨﻛﺎﺑﻪﺴﻳﺎﻘﻣﺭﺩﻩﺪﺷﺭﺎﻤﻴﺗ K562ﻱﺎﻫﻝﻮﻠﺳﺭﺩ p210BCR-ABL
ﺪﻧﺎﺑ،ﻥﺁﺎﺑﻥﺎﻣﺯﻢﻫﻰﻟﻭ .(5 ﻞﻜﺷ) ﺪﻳﺩﺮﮔﻮﺤﻣﺖﻋﺎﺳ 48 ﻭ 24 ﺯﺍﺲﭘ
ﺯﺍﺲﭘﺐﻳﺮﻘﺗﻪﺑﻭﻪﺘﺸﮔﻒﻴﻌﺿﺖﻋﺎﺳ 24 ﺕﺪﻣﺭﺩﻝﺎﻣﺮﻧp160BCR
ﻡﺎﻤﺗﺭﺩp83BCR related proteinﻪﺑﻁﻮﺑﺮﻣﺪﻧﺎﺑ .(5 ﻞﻜﺷ) ﺪﺷﻮﺤﻣﺖﻋﺎﺳ 48
.(6 ،5 ﻞﻜﺷ) ﺩﺮﻜﻧﺮﻴﻴﻐﺗﺖﻋﺎﺳ 48 ﻭ 24 ﺯﺍﺲﭘﺎﻫﻩﻭﺮﮔ
5 ﺖﻈﻠﻏ ﺕﺭﻭﺎﺠﻣ ﺭﺩ K562 ﻱﺎﻫﻝﻮﻠﺳ ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻰﺑﺎﻳﺯﺭﺍ
ﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 7 ﻩﺍﺮﻤﻫﻪﺑﺲﻨﺳﻰﺘﻧﺁﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ
ﻭﺩﺮﻫﻪﻛﺩﺍﺩﻥﺎﺸﻧﺖﻋﺎﺳ 24 ﺕﺪﻣﻪﺑﺖﻴﻜﻴﻠﭙﻳﺮﺗﺕﺭﻮﺻﻪﺑﺮﺗﺭﻮﭘﻭﺪﻧﺍ
ﻑﺮﻇﻰﻔﻨﻣﻝﺮﺘﻨﻛﺎﺑﻪﺴﻳﺎﻘﻣﺭﺩBCRﻭBCR-ABLﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﺪﻧﺎﺑ
.(6 ﻞﻜﺷ) ﺪﺷﻮﺤﻣﻞﻣﺎﻛﺭﻮﻃﻪﺑﺖﻋﺎﺳ 24 ﺕﺪﻣ
ﺖﻈﻠﻏﺭﻮﻀﺣﺭﺩJurkatﻯﺎﻫﻝﻮﻠﺳﺕﻼﺑﻥﺮﺘﺳﻭﺞﻳﺎﺘﻧﻩﺪﻫﺎﺸﻣ ﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 7 ﻭ ﺲﻨﺳﻰﺘﻧﺁﺯﺍ ﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑ ﺮﺘﻣﻭﺮﻜﻴﻣ 2
ﺭﺩ،ﺪﻳﺩﺮﮔﻮﺤﻣﺖﻋﺎﺳ 48 ﻑﺮﻇBCRﺪﻧﺎﺑﻪﻛﺩﺍﺩﻥﺎﺸﻧﺮﺗﺭﻮﭘﻭﺪﻧﺍ ﺭﺎﻤﻴﺗﻰﻔﻨﻣﻝﺮﺘﻨﻛﻩﻭﺮﮔﺭﺩp83BCR related proteinﻪﺑﻁﻮﺑﺮﻣﺪﻧﺎﺑﻪﻛﻰﻟﺎﺣ
ﺲﻨﺳﻰﺘﻧﺁﺎﺑﻩﺪﺷﺭﺎﻤﻴﺗﻰﻠﺧﺍﺩﻝﺮﺘﻨﻛﻭﻰﻔﻨﻣﻝﺮﺘﻨﻛﺲﻨﺳﻰﺘﻧﺁﺎﺑﻩﺪﺷ
.(7 ﻞﻜﺷ) ﺩﻮﺑﺩﻮﻬﺸﻣ
ﺯﻮﺘﭘﻮﭘﺁﺮﻈﻧﺯﺍk562ﻱﺎﻫﻝﻮﻠﺳﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ (B .ﺖﻋﺎﺳ 24 ﺯﺍﺪﻌﺑﺯﻮﺘﭘﻮﭘﺁﺮﻈﻧﺯﺍk562ﻱﺎﻫﻝﻮﻠﺳﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ(A :3 ﻞﻜﺷ
ﻱﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ(C .ﺖﺷﺍﺩﻰﻟﻭﺰﻧﺮﻴﺳﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺖﻈﻠﻏﺶﻳﺍﺰﻓﺍﺎﺑﺎﻫﻝﻮﻠﺳﺯﻮﺘﭘﻮﭘﺁﻥﺍﺰﻴﻣ،ﺖﻋﺎﺳ 48 ﺯﺍﺪﻌﺑ
.ﺩﺍﺪﻧﻥﺎﺸﻧﻯﺮﻴﻴﻐﺗﻲﻔﻨﻣﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋﻪﺑﺖﻋﺎﺳ 48 ﺯﺍﺪﻌﺑﺯﻮﺘﭘﻮﭘﺁﺮﻈﻧﺯﺍJurkatﻱﺎﻫﻝﻮﻠﺳ
BCR
BCR
BCR
BCR-ABL
BCR BCR-ABL
ﻱﺎﻫﻝﻮﻠﺳﻪﻬﺟﺍﻮﻣﺯﺍﺪﻌﺑﺕﻼﺑ ﻥﺮﺘﺳﻭﺯﺍﻞﺻﺎﺣﺞﻳﺎﺘﻧ (A :5 ﻞﻜﺷ
ﺲﻨﺳﻲﺘﻧﺁ ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑ ﺭﻻﻮﻣﻭﺮﻜﻴﻣ 2 ﺎﺑ k562
ﺩﻮﻬﺸﻣﻞﻜﺷ ﺯﺍﻪﻛﺭﻮﻃ ﻥﺎﻤﻫ .ﺪﻫﺩﻰﻣ ﻥﺎﺸﻧ ﺍﺭﺖﻋﺎﺳ 24 ﺕﺪﻣﻪﺑ
ﻩﺪﻳﺩ ﺖﻋﺎﺳ 24ﺯﺍ ﺲﭘ p210BCR-ABL ﻦﻴﺌﺗﻭﺮﭘ ﻥﺎﻴﺑ ﺯﺍ ﻯﺮﺛﺍ ﺖﺳﺍ
ﻥﺎﻴﺑ ﻭ ﺖﺳﺍ ﻩﺪﺷ ﺮﺗﻒﻴﻌﺿ p160BCR ﻥﺎﻴﺑ ﻦﻴﻨﭽﻤﻫ .ﺩﻮﺷﻰﻤﻧ
ﺞﻳﺎﺘﻧ(B .ﺖﺳﺍﻩﺩﺮﻜﻧﻯﺮﻴﻴﻐﺗﻲﻠﺧﺍﺩﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋﻪﺑp83BCR related protein
ﺭﻻﻮﻣﻭﺮﻜﻴﻣ 2 ﺎﺑ k562ﻱﺎﻫﻝﻮﻠﺳﻪﻬﺟﺍﻮﻣﺯﺍﺪﻌﺑﺕﻼﺑﻥﺮﺘﺳﻭﺯﺍﻞﺻﺎﺣ
ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺖﻋﺎﺳ 48 ﺯﺍﺲﭘﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺮﺘﻴﻟﻞﻴﻣﺮﺑ
ﻥﺎﻴﺑ .ﺖﺳﺍﻪﺘﻓﺭﻦﻴﺑﺯﺍ p160BCRﻭp210BCR-ABLﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﻥﺎﻴﺑﻪﻛ
ﺖﺳﺍﻩﺩﺮﻜﻧﻯﺮﻴﻴﻐﺗﻲﻠﺧﺍﺩﻝﺮﺘﻨﻛﻥﺍﻮﻨﻋﻪﺑp83BCR
.
BCR
BCR
k562ﻱﺎﻫﻝﻮﻠﺳﻪﻬﺟﺍﻮﻣﺯﺍﺪﻌﺑﺕﻼﺑﻥﺮﺘﺳﻭﺯﺍﻞﺻﺎﺣﺞﻳﺎﺘﻧ :6 ﻞﻜﺷ 24 ﺕﺪﻣﻪﺑﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺮﺘﻴﻟﻲﻠﻴﻣﺮﺑﺭﻻﻮﻣﻭﺮﻜﻴﻣ 5 ﺎﺑ
ﺮﻫ .p83BCRrelated proteinﻭp210BCR-ABL،p160BCRﻥﺎﻴﺑﺮﻈﻧﺯﺍﺖﻋﺎﺳ
ﺭﻮﻃﻪﺑﺖﻋﺎﺳ 24 ﺕﺪﻣﻑﺮﻇBCRﻭBCR-ABLﻯﺎﻫﻦﻴﺌﺗﻭﺮﭘﺪﻧﺎﺑﻭﺩ
.ﺪﺷﻮﺤﻣﻞﻣﺎﻛ
ﺎﻫﻝﻮﻠﺳﻥﺪﻧﺎﻣﻩﺪﻧﺯﻯﻭﺭﺮﺑﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺮﻴﺛﺎﺗ
ﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 2 ﺎﺑﻩﺪﺷﺭﺎﻤﻴﺗﻭﻝﺎﻣﺮﻧﻩﻭﺮﮔﻭﺩﺭﺩ K562ﻱﺎﻫﻝﻮﻠﺳ ﺕﺪﻣﻪﺑﺮﺗﺭﻮﭘﻭﺪﻧﺍﺭﻻﻮﻣﻲﻠﻴﻣﺮﺑﺮﺘﻣﻭﺮﻜﻴﻣ 7 ﻭﺲﻨﺳﻰﺘﻧﺁﺯﺍﺭﻻﻮﻣﻲﻠﻴﻣ
ﻩﺩﺍﺩﺖﺸﻛ،FBSﺪﺻﺭﺩ10 ﻩﺍﺮﻤﻫﻪﺑRPMIﻂﻴﺤﻣﺭﺩﺖﻋﺎﺳ 192
ﻭﺩﺮﻫﻯﺍﺮﺑﻭﺪﺷﺵﺭﺍﺰﮔﺖﻋﺎﺳ 24 ﺮﻫﻩﺪﻧﺯﻯﺎﻫﻝﻮﻠﺳﺩﺍﺪﻌﺗ .ﺪﻧﺪﺷ
ﺎﻫﻝﻮﻠﺳﺮﻴﺜﻜﺗﺩﻮﺷﻰﻣﻩﺪﻫﺎﺸﻣﻪﻛﺭﻮﻃﻥﺎﻤﻫ .ﺪﺷﻢﺳﺭﺭﺍﺩﻮﻤﻧﻩﻭﺮﮔ
ﺪﺻﺭﺩ .ﺖﺳﺍﺭﺎﻤﻴﺗﻥﻭﺪﺑﻩﻭﺮﮔﺯﺍﺮﺗﻦﻴﻳﺎﭘﺭﺎﻴﺴﺑﻩﺪﺷﺭﺎﻤﻴﺗﻩﻭﺮﮔﺭﺩ
ﺖﺷﺍﺩﻰﻟﻭﺰﻧﺮﻴﺳ،ﻥﺎﻣﺯﺖﺷﺬﮔﺎﺑﺭﺎﻤﻴﺗﻥﻭﺪﺑﻩﻭﺮﮔﺭﺩﻩﺪﻧﺯﻯﺎﻫﻝﻮﻠﺳ ﻰﻤﻈﻨﻣﺎﻧﻯﻮﮕﻟﺍﺯﺍﻥﺎﻣﺯﺖﺷﺬﮔﺎﺑﻩﺪﺷﺭﺎﻤﻴﺗﻩﻭﺮﮔﺭﺩﺪﺻﺭﺩﻦﻳﺍﻰﻟﻭ
24 ﺯﺍﺲﭘﺪﺻﺭﺩ 67 ﺯﺍﻥﺪﻧﺎﻣﻩﺪﻧﺯ ﺪﺻﺭﺩﻪﭼﺮﮔﺍ .ﺖﺳﺍﺭﺍﺩﺭﻮﺧﺮﺑ
ﺪﻴﺳﺭﺖﻋﺎﺳ 192 ﻰﻃﺪﺻﺭﺩ19 ﻪﺑﺲﻨﺳﻰﺘﻧﺁﻮﻨﻴﻟﻮﻓﺭﻮﻣﺎﺑﺭﺎﻤﻴﺗﺖﻋﺎﺳ
p≤0/01 ﺎﺑﺭﺍﺩﻰﻨﻌﻣﺭﻮﻃﻪﺑ (ﺪﺻﺭﺩ 63) ﻝﺮﺘﻨﻛﻩﻭﺮﮔﺎﺑﻪﺴﻳﺎﻘﻣﺭﺩﻪﻛ
.ﺖﺷﺍﺩﺶﻫﺎﻛ
ﻪﺑﻁﻮﺑﺮﻣﺪﻧﺎﺑ :ﺖﻋﺎﺳ 48 ﺯﺍﺲﭘJurkatﻯﺎﻫﻝﻮﻠﺳﺕﻼﺑﻥﺮﺘﺳﻭ :7 ﻞﻜﺷ
ﻝﺮﺘﻨﻛﺲﻨﺳﻰﺘﻧﺁﺎﺑﻩﺪﺷﺭﺎﻤﻴﺗ) ﻰﻔﻨﻣﻝﺮﺘﻨﻛﻩﻭﺮﮔﺭﺩp83BCR related protein
.ﺖﺳﺍﺩﻮﻬﺸﻣ (ﺲﻨﺳﻰﺘﻧﺁﺎﺑﻩﺪﺷﺭﺎﻤﻴﺗ) ﻰﻠﺧﺍﺩﻝﺮﺘﻨﻛﻭ (ﻰﻔﻨﻣ
ﺚﺤﺑ
ﻦﻴﺌﺗﻭﺮﭘ ﻯﻭﺭ ﺲﻨﺳﻲﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﺮﻴﺛﺎﺗ ،ﻪﻌﻟﺎﻄﻣ ﻦﻳﺍ ﺭﺩ ﻥژ ﻞﻣﺎﺣ K562 ﻯﺎﻫﻝﻮﻠﺳ ﻥﺪﻧﺎﻣ ﻩﺪﻧﺯ ﺪﺻﺭﺩ ﻭ BCR-ABL ﻪﻛﺩﺍﺩ ﻥﺎﺸﻧﺕﻼﺑ ﻥﺮﺘﺳﻭﺯﺍﻞﺻﺎﺣ ﺞﻳﺎﺘﻧ .ﺪﺷﻰﺑﺎﻳﺯﺭﺍ BCR-ABL
،ﺪﺸﻧﻥﺎﻴﺑﺖﻋﺎﺳ 24 ﺯﺍﺲﭘﺲﻨﺳﻰﺘﻧﺁﺭﻮﻀﺣﺭﺩBCR-ABL ﻦﻴﺌﺗﻭﺮﭘ ﻦﻴﺌﺗﻭﺮﭘﻥﺎﻴﺑ،ﺲﻨﺳﻰﺘﻧﺁﻦﻴﻳﺎﭘﺖﻈﻠﻏﺭﻮﻀﺣﺭﺩﻥﺁﺎﺑﻥﺎﻣﺯﻢﻫﻪﭼﺮﮔﺍ
.ﺩﻭﺭﻰﻣﻦﻴﺑﺯﺍﺖﻋﺎﺳ 48 ﺯﺍﺲﭘﺖﻳﺎﻬﻧﺭﺩﻭﻩﺪﺷﻒﻴﻌﺿﻢﻫBCR
.ﺪﻧﺍﻩﺩﺍﺩﻡﺎﺠﻧﺍﻰﻤﺳﻮﻟﻥﺎﻣﺭﺩﺖﻬﺟﺭﺩﻯﺩﺎﻳﺯﻯﺎﻫﺵﻼﺗ،ﻦﻴﻘﻴﻘﺤﻣ
ﻦﻴﻟﻭﺍﻥﺍﻮﻨﻋﻪﺑ ﺐﻴﻨﻴﺗﺎﻤﻳﺍﺯﺍﻩﺩﺎﻔﺘﺳﺍ،ﺎﻫﻥﺎﻣﺭﺩﻦﻳﺍﻦﻳﺮﺗﺞﻳﺍﺭﺯﺍﻰﻜﻳ ﻦﻳﺍ .ﺪﻳﺩﺮﮔ ﺡﺮﻄﻣ ﻚﻴﻨﻴﻠﻛ ﺭﺩ ﻪﻛ ﺩﻮﺑ ﺯﺎﻨﻴﻛ ﻦﻳﺯﻭﺮﻴﺗ ﻩﺪﻨﻨﻛﺭﺎﻬﻣ
ﺭﺩ(Adenosine Tri Phosphate)ATPﻝﺎﺼﺗﺍﻩﺎﮕﻳﺎﺟﻩﺪﻨﻨﻛﺭﺎﻬﻣ ﻝﻮﻜﻟﻮﻣﻝﺎﻌﻓﻡﺮﻓﻞﻴﻜﺸﺗﺯﺍﻭﺪﻨﻛﻰﻣﻝﺎﻐﺷﺍﺍﺭABL-Kinaseﻦﻴﻣﻭﺩ
ﺍﺭﻰﺗﻼﻜﺸﻣﻥﺁﺯﺍﻩﺩﺎﻔﺘﺳﺍﻰﻟﻭ .ﺪﻳﺎﻤﻧﻰﻣﻯﺮﻴﮔﻮﻠﺟﻰﻳﺎﻀﻓﺖﻟﺎﺣﺭﺩ
ﻰﻣﺯﻮﻣﻭﺮﻛﻯﺎﻫﻯﺭﺎﺠﻨﻫﺎﻧ،BCR-ABLﻥﺎﻴﺑﺶﻳﺍﺰﻓﺍ :ﺪﻨﻧﺎﻣﺖﺷﺍﺩﻰﭘﺭﺩ
ﻦﻴﻣﻭﺩ ﻯﺎﻫﻥﻮﻴﺳﺎﺗﻮﻣ ،ﺖﺒﺜﻣ ﺎﻴﻔﻟﺩﻼﻴﻓ ﻯﺎﻫﻝﻮﻠﺳ ﺖﻴﻌﻤﺟ ﺭﺩ ﻰﻓﺎﺿﺍ ﺚﻋﺎﺑﻩﺪﺷﺮﻛﺫﻞﻣﺍﻮﻋ .(18) Oct-1ﻯﺎﻫﻢﺴﻴﻓﺭﻮﻣﻰﻠﭘﻭﺯﺎﻨﻴﻛABL
.ﺪﻨﺷﺎﺑ ﻯﺮﮕﻳﺩﻦﻳﺰﮕﻳﺎﺟ ﻯﺎﻫﻭﺭﺍﺩﻦﺘﻓﺎﻳﺩﺪﺻﺭﺩﻦﻴﻘﻘﺤﻣ ﺎﺗﺪﻳﺩﺮﮔ
BCR
ﻰﭘﺍﺮﺗﻥژﺭﺍﻩﺩﺎﻔﺘﺳﺍ،ﺪﺷﻰﻣﻩﺩﺍﺩﻪﻨﻴﻣﺯﻦﻳﺍﺭﺩﻪﻛﻰﻳﺎﻫﺪﻴﻣﺍﺯﺍﻰﻜﻳ
BCR-ABL ﻥژ ﻥﺩﺮﻛ ﺵﻮﻣﺎﺧ ﺯﺍﻞﺻﺎﺣ ﺞﻳﺎﺘﻧ ﺎﻣﺍ .ﺩﻮﺑ siRNAﺎﺑ
ﺯﺍﺕﺎﻘﻴﻘﺤﺗﻦﻳﺍﻰﻣﺎﻤﺗﻪﻛﻰﻟﺎﺣﺭﺩﺩﻮﺑﺾﻴﻘﻧﻭﺪﺿ siRNAﻂﺳﻮﺗ ﻥﺍﻮﻨﻋﻪﺑK562ﻝﻮﻠﺳﺯﺍﻰﮕﻤﻫﻭﺪﻧﺩﻮﺑﻰﻧﺎﺴﻜﻳ siRNAﻯﺎﻫﻰﻟﺍﻮﺗ ،ﻩﺪﻣﺁﺖﺳﺩﻪﺑﺞﻳﺎﺘﻧﺯﺍ .ﺪﻧﺩﻮﺑﻩﺩﺮﻛﻩﺩﺎﻔﺘﺳﺍﺖﺒﺜﻣBCR-ABLﻝﺪﻣ
ﺮﺑ siRNAﺩﻭﺭﻭﺯﺍﺖﻋﺎﺳ 48 ﺯﺍﺲﭘﺎﻫﻝﻮﻠﺳﻪﻛﺪﻳﺩﺮﮔﺺﺨﺸﻣ
siRNAﺕﺪﻣﻦﻳﺍﺯﺍﺪﻌﺑﺎﻣﺍﺪﻧﻮﺷﻲﻣﺯﻮﺘﭘﻮﭘﺁﺯﺎﻓﺩﺭﺍﻭBCR-ABLﻪﻴﻠﻋ
ﺯﺎﺑﺍﺭﺩﻮﺧﺕﺎﻴﺣﺎﻫﻝﻮﻠﺳﺩﺪﺠﻣﺭﻮﻃﻪﺑﻭﻩﺪﻳﺩﺮﮔﻰﻣﺰﻧﺁﺐﻳﺮﺨﺗﺭﺎﭼﺩ CMLﻰﻧﺎﻣﺭﺩﻥژﻯﺍﺮﺑsiRNAﺯﺍﻩﺩﺎﻔﺘﺳﺍﻦﻳﺍﺮﺑﺎﻨﺑ .(19-22) ﺪﻨﺑﺎﻳﻰﻣ
ﻯﺮﮕﻳﺩﻚﻴﺘﺘﻨﺳﻯﺎﻫﺲﻨﺳﻰﺘﻧﺁﻭﺭﻦﻳﺍﺯﺍ .ﺪﺷﺎﺑﺐﺳﺎﻨﻣﺵﻭﺭﺪﻧﺍﻮﺗﻰﻤﻧ
ﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺎﻬﻧﺁﺯﺍﻰﻜﻳﻪﻛﺖﻓﺎﻳﻞﻣﺎﻜﺗﻥﺎﻘﻘﺤﻣﻂﺳﻮﺗ ﺪﺟﺍﻭﻪﺑﺎﺸﻣﻯﺎﻫﺲﻨﺳﻰﺘﻧﺁﻪﺑﺖﺒﺴﻧﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ .ﺩﻮﺑ
ﻪﺑﻪﺘﺴﺑﺍﻭﻉﺍﻮﻧﺍﻑﻼﺧﺮﺑﺲﻨﺳﻰﺘﻧﺁﻦﻳﺍ؛ﺪﺷﺎﺑﻰﻣﻯﺭﺎﻴﺴﺑﺕﻮﻗﻁﺎﻘﻧ ﻞﻴﻟﺩﻪﺑﻭﻪﺘﺷﺍﺩﻑﺪﻫﻰﻟﺍﻮﺗﻪﺑﺖﺒﺴﻧﻯﺮﺗﻻﺎﺑﺹﺎﺼﺘﺧﺍ RNaseH ﺩﻮﺟﻮﻣ ﻯﺎﻫﻢﻳﺰﻧﺁﻉﺍﻮﻧﺍﻰﻣﺎﻤﺗﻪﺑ ﺩﺭﺍﺩﺩﻮﺧﺭﺎﺘﺧﺎﺳ ﺭﺩﻪﻛﻰﺗﺍﺮﻴﻴﻐﺗ ﺖﺳﺍﻩﺪﺸﻧﺖﻓﺎﻳﻰﻤﻳﺰﻧﺁﺰﻴﻧﻝﺎﺣﻪﺑﺎﺗﻭﺖﺳﺍﻡﻭﺎﻘﻣﺎﻫﻝﻮﻠﺳﻥﻭﺭﺩﺭﺩ ﺍﺭﻥﺎﻜﻣﺍﻦﻳﺍﺎﻫﺖﻴﺻﻮﺼﺧﻦﻳﺍ .(16) ﺪﻳﺎﻤﻧﺐﻳﺮﺨﺗﺍﺭﻥﺁﺪﻧﺍﻮﺘﺑﻪﻛ
،ﺖﺸﻛﻂﻴﺤﻣﺭﺩﻰﻧﻻﻮﻃﺕﺪﻣﻪﺑﺲﻨﺳﻰﺘﻧﺁﻦﻳﺍﺯﺍﻢﻴﻧﺍﻮﺘﺑﺎﺗﺪﻫﺩﻰﻣ ﻦﻳﺍ .ﻢﻴﻳﺎﻤﻧﻩﺩﺎﻔﺘﺳﺍﻥﺁﺮﺛﺍﺶﻫﺎﻛﺎﻳﺐﻳﺮﺨﺗﻥﺎﻜﻣﺍﺯﺍﻥﺩﻮﺑﻥﺍﺮﮕﻧﻥﻭﺪﺑ
ﺯﺍﻥﺍﻮﺗﻰﻣﻦﻳﺍﺮﺑﺎﻨﺑﺪﻳﺎﻤﻧﻰﻤﻧﺐﻳﺮﺨﺗﺍﺭﺩﻮﺧﻑﺪﻫﻰﻟﺍﻮﺗﺲﻨﺳﻰﺘﻧﺁ ﻩﺩﺎﻔﺘﺳﺍﻰﻟﻮﻠﺳﻯﺎﻫﺩﺍﺪﺧﺭﻭﻚﻳژﻮﻟﻮﻴﺑﻯﺎﻫﻩﺪﻳﺪﭘﺮﺘﺸﻴﺑﻙﺭﺩﺭﺩﻥﺁ ﻭﻩﺩﻮﻤﻧﺐﻳﺮﺨﺗﺍﺭﻑﺪﻫﻰﻟﺍﻮﺗRNaseHﻪﺑﻪﺘﺴﺑﺍﻭﻉﺍﻮﻧﺍﺍﺮﻳﺯ؛ﺩﻮﻤﻧ ﺕﺍﺮﻴﺛﺎﺗﺖﺳﺍﻦﻜﻤﻣﻪﻛﺪﻳﺎﻤﻧﻰﻣﺎﻫﺭﻝﻭﺯﻮﺘﻴﺳﺭﺩﺍﺭﻥﺁﺯﺍﻞﺻﺎﺣﺩﺍﻮﻣ ﺮﺑﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ .(23) ﺩﺭﺍﺬﮕﺑﻯﺎﺟﻪﺑﺍﺭﻯﺍﻩﺪﺸﻧﻰﻨﻴﺑﺶﻴﭘ
ﺩﻮﺷﻰﻣﺚﻋﺎﺑﺖﻴﺻﻮﺼﺧﻦﻳﺍﻭﺖﺳﺍﻰﻧﻮﻳﺮﻴﻏﻰﺒﻴﻛﺮﺗ،RNAiﻑﻼﺧ ﺎﻫﻝﻮﻠﺳﻞﺧﺍﺩﻪﺑﻥﺁﻯﺭﻮﻴﻟﺩﻯﺍﺮﺑﺮﺗﺭﻮﭘﻭﺪﻧﺍﻯﺭﻮﻴﻟﺩﺪﻴﺘﭙﭘﺯﺍﻥﺍﻮﺘﺑﺎﺗ ﻪﺑﻰﻬﺟﻮﺗﻞﺑﺎﻗﺖﻳﺰﻣﺩﻮﺧﻪﻛﺩﻮﻤﻧﻩﺩﺎﻔﺘﺳﺍﻰﺘﻴﺴﻴﺴﻛﻮﺗﺩﺎﺠﻳﺍﻥﻭﺪﺑ
.(24) ﺪﻳﺁﻰﻣﺏﺎﺴﺣ
ﺵﻮﻣﺎﺧﻯﺍﺮﺑﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻯﺎﻫﺖﻳﺩﻭﺪﺤﻣﺯﺍﻰﻜﻳ ﻦﻳﺍﺎﺑ .ﺖﺳﺍﻥﻭﺪﻛﺕﺭﺎﺘﺳﺍﻰﻟﺍﻮﺗﻪﺑﺎﻬﻧﺁﻥﺩﻮﺑﺩﻭﺪﺤﻣﺎﻫﻥژﻥﺩﺮﻛ
ﺵﻮﻣﺎﺧﻯﺍﺮﺑﺲﻨﺳﻰﺘﻧﺁﻦﻳﺍﺯﺍﻩﺩﺎﻔﺘﺳﺍﻪﻛﺩﺩﺮﮔﻰﻣﺺﺨﺸﻣﻒﻴﺻﻮﺗ ﻪﻛﺩﻮﺷﻰﻣﻝﺎﻣﺮﻧBCRﻥژ ﻰﺷﻮﻣﺎﺧﺚﻋﺎﺑ BCR-ABL ﻥژﻥﺩﺮﻛ
ﻥﻮﻴﺳﺎﻛﻮﻠﺴﻧﺍﺮﺗ ﺭﺩ ﻰﻠﻟﺍ ﻑﺬﺣ ﺩﻮﺟﻭ ﻡﺪﻋ ﻞﻴﻟﺩ ﻪﺑ ﻩﺪﻳﺪﭘ ﻦﻳﺍ ﻪﺘﻓﺎﻳﻡﺎﻏﺩﺍﻯﺎﻫﻥژﻉﺍﻮﻧﺍﻡﺎﻤﺗﻰﻓﺮﻃﺯﺍ .(25) ﺪﻫﺩﻰﻣﺥﺭﺎﻴﻔﻟﺩﻼﻴﻓ
ﻼﻣﺎﻛﻥﻭﺪﻛﺕﺭﺎﺘﺳﺍﻪﻴﺣﺎﻧﺭﺩ (ﻩﺮﻴﻏﻭb2/a2ﻭ b3/a, BCR-ABL)
ﺕﺭﺎﺘﺳﺍ ﻪﻴﻠﻋ ﺲﻨﺳﻰﺘﻧﺁ ﻦﻳﺍ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﻦﻳﺍﺮﺑﺎﻨﺑ .ﺪﺷﺎﺑﻰﻣ ﻥﺎﺴﻜﻳ
ﺭﺎﻛﻪﺑ BCR-ABLﻉﺍﻮﻧﺍﻡﺎﻤﺗﻥﺩﺮﻛﺵﻮﻣﺎﺧﻯﺍﺮﺑﺪﻧﺍﻮﺗﻰﻣﻥﻭﺪﻛ
ﺖﻣﻭﺎﻘﻣ ﻩﺪﻳﺪﭘ ﻪﺑ ﺮﺠﻨﻣﻪﻛ ﻰﻳﺎﻫﺶﻬﺟ ﻡﺎﻤﺗ،ﻦﻴﻨﭽﻤﻫ .(25) ﺩﻭﺭ
ABL ﻯﺯﺎﻨﻴﻛﻦﻣﻭﺩ ﺭﺩﻰﺗﺍﺮﻴﻴﻐﺗ ﻪﺑ ﻁﻮﺑﺮﻣﺪﻧﺩﺮﮔﻰﻣﺐﻴﻨﻴﺗﺎﻤﻳﺍﻪﺑ
ﻥﻭﺪﻛﺕﺭﺎﺘﺳﺍﻪﻴﻠﻋﻩﺪﺷﻰﺣﺍﺮﻃﺲﻨﺳﻲﺘﻧﺁﺪﻧﺰﮔﺯﺍﻭﺭﻦﻳﺍﺯﺍ،ﺖﺳﺍ ﻥﺪﻴﺳﺭﻪﻌﻟﺎﻄﻣﻦﻳﺍﻑﺍﺪﻫﺍﺯﺍﻲﻜﻳ .(26 ،18) ﺩﻮﺑﺪﻨﻫﺍﻮﺨﻧﻥﺎﻣﺍﺭﺩ .ﺖﺳﺍﻩﺩﻮﺑﺯﻮﺘﭘﻮﭘﺁ ﺶﻳﺍﺰﻓﺍﻭ ﻰﻟﻮﻠﺳﺮﻴﺜﻜﺗﺭﺎﻬﻣﻥﺍﺰﻴﻣ ﺮﺜﻛﺍﺪﺣﻪﺑ
ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﻪﻨﻴﻬﺑﺖﻈﻠﻏ،ﺭﻮﻛﺬﻣﻑﺪﻫﻪﺑﻥﺪﻴﺳﺭﻯﺍﺮﺑﻦﻳﺍﺮﺑﺎﻨﺑ ﻑﻼﺧ ﺮﺑ .ﺪﺷﻦﻴﻴﻌﺗﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﻚﻴﻨﻜﺗﺯﺍﻩﺩﺎﻔﺘﺳﺍﺎﺑﺲﻨﺳﻰﺘﻧﺁ
ﺭﺩ) ﺲﻨﺳﻰﺘﻧﺁ ﺖﻈﻠﻏﺶﻳﺍﺰﻓﺍ ﺎﺑ ﻪﻛﺪﻳﺩﺮﮔ ﺺﺨﺸﻣ ،ﻪﻴﻟﻭﺍ ﺭﺎﻈﺘﻧﺍ
ﻰﻟﻮﻠﺳﺮﻴﺜﻜﺗﺯﺍﺖﻌﻧﺎﻤﻣﻥﺍﺰﻴﻣ (ﺮﺘﻴﻟﻰﻠﻴﻣﺮﺑﻝﻮﻣﻭﺮﻜﻴﻣ 2-5 ﻩﺩﻭﺪﺤﻣ
ﺺﺨﺸﻣ ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻡﺎﺠﻧﺍ ﺯﺍ ﺲﭘ .ﺖﺷﺍﺩ ﻰﻟﻭﺰﻧﺮﻴﺳ ﺯﻮﺘﭘﻮﭘﺁ ﻭ
ﻝﻮﻣﻭﺮﻜﻴﻣ 2) ﺲﻨﺳﻰﺘﻧﺁ ﻪﻨﻴﻬﺑ ﺖﻈﻠﻏ ﺎﺑ ﻪﻛ ﻰﻳﺎﻫﻝﻮﻠﺳ ﻪﻛ ﺪﺷ
ﻦﻴﺌﺗﻭﺮﭘ ،ﺪﻧﺩﻮﺑ ﻩﺪﺷ ﺭﺎﻤﻴﺗ (ﺖﺸﻛ ﻂﻴﺤﻣ ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﻫ ﻱﺍﺯﺍ ﻪﺑ
ﺭﺩBCRﻦﻴﺌﺗﻭﺮﭘ ﻭ ﻩﺪﻳﺩﺮﮔﻑﺬﺣ ﺖﻋﺎﺳ 24 ﻑﺮﻇBCR-ABL
.ﺪﺷ ﻑﺬﺣ ﺖﻋﺎﺳ 48 ﺯﺍ ﺲﭘ ﻭ ﻩﺪﺷ ﻒﻴﻌﻀﺗ ﺖﻋﺎﺳ 24 ﺕﺪﻣ
5 ﺖﻈﻠﻏ ﺎﺑ ﻩﺪﺷﺭﺎﻤﻴﺗ ﻯﺎﻫﻝﻮﻠﺳ ﻯﺍﺮﺑﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻪﻜﻨﻳﺍﺯﺍ ﺲﭘ
ﻥژﻭﺩﺮﻫﻪﻛﺪﺷﺺﺨﺸﻣ،ﺪﻳﺩﺮﮔﺭﺍﺮﻜﺗﺲﻨﺳﻰﺘﻧﺁﺯﺍﺭﻻﻮﻣﻭﺮﻜﻴﻣ ﻞﻣﺎﻛﺭﻮﻃﻪﺑﺐﻳﺮﻘﺗﺭﻮﻃﻪﺑﺖﻋﺎﺳ 24 ﻑﺮﻇBCRﻭBCR-ABL
ﺶﻳﺍﺰﻓﺍﻪﺑ ﺮﺠﻨﻣﺲﻨﺳﻰﺘﻧﺁ ﺖﻈﻠﻏﺶﻳﺍﺰﻓﺍﻦﻴﻨﭽﻤﻫ .ﺪﻧﺪﺷﺵﻮﻣﺎﺧ
،ﺪﻨﻛﻰﻣﻪﻴﺟﻮﺗ ﺍﺭﻩﺪﻳﺪﭘ ﻦﻳﺍ ﻪﻛﻪﭽﻧﺁﻦﻳﺍﺮﺑﺎﻨﺑ .ﺪﺸﻧﺯﻮﺘﭘﻮﭘﺁ ﻥﺍﺰﻴﻣ
ﻯﻻﺎﺑ ﻯﺎﻫﺖﻈﻠﻏ ﺭﺩ .ﺖﺳﺍ ﺯﻮﺘﭘﻮﭘﺁ ﺪﻨﻳﺍﺮﻓ ﻦﻳﺍ ﺭﺩBCR ﺖﻟﺎﺧﺩ BCR-ﻥژﻭﺩﺮﻫﻊﻳﺮﺳﻰﺷﻮﻣﺎﺧﻞﻴﻟﺩﻪﺑﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ
ﺭﺩﺩﺩﺮﮔﻰﻣﻒﻴﻌﻀﺗﺯﻮﺘﭘﻮﭘﺁﻱﺎﻘﻟﺍ،ﺖﻋﺎﺳ 24 ﻑﺮﻇBCRﻭABL
ﺮﺗﻯﻮﻗBCRﻥﺎﻴﺑﻥﻮﭼ) ﺲﻨﺳﻰﺘﻧﺁﻦﻴﻳﺎﭘﻯﺎﻫﺖﻈﻠﻏﺯﺍﻪﻛﻰﺗﺭﻮﺻ
ﺭﺍﺩﺍﻭﺮﺘﺸﻴﺑﺍﺭﻝﻮﻠﺳﻭﺪﺷﺵﻮﻣﺎﺧﻥﺁﺯﺍﺮﺗﺮﻳﺩ (ﺖﺳﺍBCR-ABLﺯﺍ
ﻪﻛ ﺪﻨﻛﻰﻣ ﻰﻋﺍﺪﺗ ﻪﻧﻮﮔ ﻦﻳﺍ ،ﻪﻠﺻﺎﺣ ﻯﺎﻫﻪﺘﻓﺎﻳ .ﺩﻮﻤﻧ ﺯﻮﺘﭘﻮﭘﺁ ﻪﺑ
ﻚﻴﺗﻮﺘﭘﻮﭘﺁﺪﺿﺮﺛﺍ ،ﺖﺴﻴﻧﻮﮔﺎﺘﻧﺁﻚﻳﻥﺍﻮﻨﻋ ﻪﺑBCR ﻥژ ﻝﻮﺼﺤﻣ
ﻝﺎﻣﺮﻧBCRﻥژ ﺶﻘﻧ،ﻦﻴﺸﻴﭘﺕﺎﻌﻟﺎﻄﻣ،ﻦﻴﻨﭽﻤﻫ .ﺩﺭﺍﺩBCR-ABL
.(26) ﺖﺳﺍﻩﺩﺮﻛﺖﺑﺎﺛ BCR-ABLﻦﻴﺌﺗﻭﺮﭘﻮﻜﻧﺁﻰﻔﻨﻣﻢﻴﻈﻨﺗﺭﺩﺍﺭ
ﻦﻴﺌﺗﻭﺮﭘﻮﻜﻧﺁ ﻂﺳﻮﺗ ﻪﻛ ﻰﻟﺎﻨﮕﻴﺳ ﻝﺎﺳﺭﺍ ﻯﺎﻫﺮﻴﺴﻣ ﻰﻓﺮﻃ ﺯﺍ ﻪﺑﻥﺁﻯﺎﻘﺑﻭﺮﻴﺜﻜﺗ،ﻰﻟﻮﻠﺳﻚﻳﺮﺤﺗﺚﻋﺎﺑ،ﺪﺘﻓﺍﻰﻣﻩﺍﺭﻪﺑBCR-ABL
.(27) ﺩﺩﺮﮔﻰﻣﺎﻫﻦﻳﺎﻛﻮﺘﻳﺎﺳﻪﺑﻪﺘﺴﺑﺍﻭﺮﻴﻏﺕﺭﻮﺻ
ﻯﺮﻴﮔ
ﻪﺠﻴﺘﻧ
ﻰﺷﻮﻣﺎﺧﻭﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺯﺍﻩﺩﺎﻔﺘﺳﺍ،ﻖﻴﻘﺤﺗﻦﻳﺍﺭﺩ ﻭﺮﻴﺜﻜﺗﺯﺍﺖﻌﻧﺎﻤﻣﻪﺑﺮﺠﻨﻣBCR-ABLﻭBCRﻥژﻭﺩﺮﻫﻥﺎﻣﺯﻢﻫ
ﺮﻤﻋﻝﻮﻃ ﻰﻨﺤﻨﻣﺭﺩ ﺮﻣﺍ ﻦﻳﺍ .ﺪﻳﺩﺮﮔK562ﻯﺎﻫﻝﻮﻠﺳ ﺪﺷﺭﻒﻗﻮﺗ ﺭﻮﻃ ﻪﺑ ﺲﻨﺳﻰﺘﻧﺁ ﺎﺑﻩﺪﺷ ﺭﺎﻤﻴﺗ ﻯﺎﻫﻝﻮﻠﺳ ﺮﺑﺍﺮﺑﺭﺩﻝﺎﻣﺮﻧﻯﺎﻫﻝﻮﻠﺳ ﺲﻨﺳﻰﺘﻧﺁﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣﺯﺍﻩﺩﺎﻔﺘﺳﺍ .(2 ﺭﺍﺩﻮﻤﻧ) ﺖﺳﺍﺺﺨﺸﻣﻞﻣﺎﻛ
،ﺖﻋﺎﺳ 192 ﺕﺪﻣﻪﺑﺖﺸﻛﻂﻴﺤﻣﺭﺩﺎﻫﻝﻮﻠﺳﺕﺪﻣﻰﻧﻻﻮﻃﻰﺳﺭﺮﺑﻭ
ﺶﻫﺎﻛﺚﻋﺎﺑﺎﻫﻝﻮﻠﺳﻥﺪﺷﻚﻴﺗﻭﺮﻜﻧﺎﻬﺘﻧﺍﺭﺩﻭﻝﻮﻠﺳﺮﻴﺜﻜﺗﺯﺍﺖﻌﻧﺎﻤﻣ ﻪﭼ K562ﻯﺎﻫﻝﻮﻠﺳﻦﺘﻓﺭﻦﻴﺑﺯﺍﻪﺑﻪﺟﻮﺗﺎﺑ .ﺪﺷﺎﻫﻝﻮﻠﺳﺮﻤﻋﻝﻮﻃ
ﻯﺍﺮﺑﺍﺭﻩﺍﺭﺪﻧﺍﻮﺗﻰﻣ،ﺮﻴﺜﻜﺗﺯﺍﺖﻌﻧﺎﻤﻣﻖﻳﺮﻃﺯﺍﻪﭼﺯﻮﺘﭘﻮﭘﺁﻖﻳﺮﻃﺯﺍ
.ﺪﻳﺎﻤﻧﺭﺍﻮﻤﻫCMLﻰﭘﺍﺮﺗﻥژﻪﻨﻴﻣﺯﺭﺩﻯﺪﻌﺑﺕﺎﻘﻴﻘﺤﺗ
100 90 80 70 60 50 40 30 20 10 0
24 48 72 96 120 144 168 192
ﺪﺻ
ﺭﺩ
ﻝﺎﻣﺮﻧ ﻩﺪﺷﺭﺎﻤﻴﺗ
ﺭﻻﻮﻣﻭﺮﻜﻴﻣ 2 ﺭﺎﻤﻴﺗﺯﺍﺲﭘﺎﻫﻝﻮﻠﺳﻥﺪﻧﺎﻣﻩﺪﻧﺯﺪﺻﺭﺩﻪﺴﻳﺎﻘﻣ :2 ﺭﺍﺩﻮﻤﻧ
ﺪﺻﺭﺩ .ﻝﺎﻣﺮﻧ ﻱﺎﻫﻝﻮﻠﺳﺎﺑ ﺲﻨﺳﻲﺘﻧﺁ ﻮﮕﻴﻟﺍﻮﻨﻴﻟﻮﻓﺭﻮﻣ ﺮﺘﻴﻟﻲﻠﻴﻣ ﺮﺑ
ﻩﺪﺷﺭﺎﻤﻴﺗﻩﻭﺮﮔﺭﺩﺭﺍﺩﻰﻨﻌﻣﺭﻮﻃﻪﺑﺖﻋﺎﺳ 192 ﺯﺍﺲﭘﻩﺪﻧﺯﻯﺎﻫﻝﻮﻠﺳ
.(p≤ 0/01) ﺖﻓﺎﻳﺶﻫﺎﻛﻮﻨﻴﻟﻮﻓﺭﻮﻣﺎﺑ
ﺮﻜﺸﺗ
ﻭ
ﺮﻳﺪﻘﺗ
ﺱﺭﺪﻣﺖﻴﺑﺮﺗ ﻩﺎﮕﺸﻧﺍﺩﺖﻧﺍﺮﮔﺯﺍ ﻩﺩﺎﻔﺘﺳﺍﺎﺑ ﻲﺗﺎﻘﻴﻘﺤﺗﺡﺮﻃ ﻦﻳﺍ ﺖﻧﻭﺎﻌﻣﺯﺍﺍﺭﺩﻮﺧﺮﻳﺪﻘﺗﻭﺮﻜﺸﺗﺪﻧﺭﺍﺩﻞﻳﺎﻤﺗﻦﻴﻔﻟﻮﻣ .ﺖﺳﺍﻩﺪﺷﻡﺎﺠﻧﺍ
ﻡﺯﻻﻱﺎﻫﻚﻤﻛﺡﺮﻃﻦﻳﺍﻡﺎﺠﻧﺍﺭﺩﻪﻛﺱﺭﺪﻣﺖﻴﺑﺮﺗﻩﺎﮕﺸﻧﺍﺩﻲﺸﻫﻭﮋﭘ
.ﺪﻧﺭﺍﺩﻡﻼﻋﺍ،ﺪﻧﺍﻪﺘﺷﺍﺩﺍﺭ
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