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Evaluation of Morpholino Antisense Oligos’ Role on BCR-ABL Gene Silencing in the K562 Cell Line

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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

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ﻪﻣﺪﻘﻣ

ﻪﻛﺪﺷﺎﺑﻰﻣﺎﻫﻲﻤﺳﻮﻟﺯﺍﻞﻜﺷﻦﻴﺘﺴﺨﻧﻦﻣﺰﻣﻱﺪﻴﻳﻮﻠﻴﻣ ﻲﻤﺳﻮﻟ ﻡﺎﻏﺩﺍﻥژﻞﻴﻟﺩﻪﺑﺮﺘﺸﻴﺑﻯﺭﺎﻤﻴﺑﻦﻳﺍﻞﻣﺎﻋ .(1-3) ﺖﺳﺍﻩﺪﺷﻲﻳﺎﺳﺎﻨﺷ

ﻪﺑﻭﻩﺪﺷﺩﺎﺠﻳﺍﺯﺎﺳﻥﻮﺧﻯﺩﺎﻴﻨﺑﻯﺎﻫﻝﻮﻠﺳﺭﺩﻪﻛﺖﺳﺍBCR-ABLﻪﺘﻓﺎﻳ

ﺎﻴﻔﻟﺩﻼﻴﻓﻡﻭﺯﻮﻣﻭﺮﻛ .(4 ،2) ﺩﻮﺷﻰﻣﻪﺘﺧﺎﻨﺷﺎﻴﻔﻟﺩﻼﻴﻓﻡﻭﺯﻮﻣﻭﺮﻛﻥﺍﻮﻨﻋ

ﻞﺑﺎﻘﺘﻣﻰﻳﺎﺟﻪﺑﺎﺟﺯﺍﻪﻛﺖﺳﺍﻩﺪﺷﻩﺎﺗﻮﻛ 22 ﻡﻭﺯﻮﻣﻭﺮﻛﻥﺎﻤﻫﻞﺻﺍﺭﺩ

ﺩﻮﺟﻭﻪﺑ (BCRﻞﻣﺎﺷ) 22 (ABLﻞﻣﺎﺷ) 9 ﻡﻭﺯﻮﻣﻭﺮﻛﺪﻨﻠﺑﻯﻭﺯﺎﺑ

ﺩﺭﻭﺁﻰﻣﺩﻮﺟﻭﻪﺑﺍﺭBCR-ABLﻪﺘﻓﺎﻳﻡﺎﻏﺩﺍﻥژ،ﻰﻳﺎﺟﻪﺑﺎﺟﻦﻳﺍ .ﺪﻳﺁﻰﻣ

ﻦﻴﺌﺗﻭﺮﭘﻮﻜﻧﺍﻪﺑﻦﻣﺰﻣ ﻱﺪﻴﻳﻮﻠﻴﻣﻲﻤﺳﻮﻟﻪﺑ ﻼﺘﺒﻣﻥﺍﺭﺎﻤﻴﺑﺮﺜﻛﺍ ﺭﺩﻪﻛ ﻝﺎﻤﺘﺣﺍﻭﺩﺯﺍﻰﻜﻳﺖﻟﺎﺣﻦﻳﺍﺭﺩ .(6 ،5) ﺩﻮﺷﻰﻣﻪﻤﺟﺮﺗ p210 BCR-ABL

ﻉﻮﻧﻭﺩ .(7) ﺩﺭﺍﺩﺩﻮﺟﻭBCRABLﻥژﻦﻴﺑb3a2b2a2 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

(3)

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

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 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

(4)

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

(5)

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

(6)

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 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

(7)

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 ﻑﺮﻇBCRBCR-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 ﻑﺮﻇBCRABL

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-ABLBCRﻥژﻭﺩﺮﻫﻥﺎﻣﺯﻢﻫ

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

ﺮﻳﺪﻘﺗ

ﺱﺭﺪﻣﺖﻴﺑﺮﺗ ﻩﺎﮕﺸﻧﺍﺩﺖﻧﺍﺮﮔﺯﺍ ﻩﺩﺎﻔﺘﺳﺍﺎﺑ ﻲﺗﺎﻘﻴﻘﺤﺗﺡﺮﻃ ﻦﻳﺍ ﺖﻧﻭﺎﻌﻣﺯﺍﺍﺭﺩﻮﺧﺮﻳﺪﻘﺗﻭﺮﻜﺸﺗﺪﻧﺭﺍﺩﻞﻳﺎﻤﺗﻦﻴﻔﻟﻮﻣ .ﺖﺳﺍﻩﺪﺷﻡﺎﺠﻧﺍ

ﻡﺯﻻﻱﺎﻫﻚﻤﻛﺡﺮﻃﻦﻳﺍﻡﺎﺠﻧﺍﺭﺩﻪﻛﺱﺭﺪﻣﺖﻴﺑﺮﺗﻩﺎﮕﺸﻧﺍﺩﻲﺸﻫﻭﮋﭘ

.ﺪﻧﺭﺍﺩﻡﻼﻋﺍ،ﺪﻧﺍﻪﺘﺷﺍﺩﺍﺭ

(8)

References

1. Goldman JM, Melo JV.Chronic myeloid leukemia--advances in biology and new approaches to treatment. N Engl J Med. 2003; 349(15): 1451-1464.

2. Faderl S, Talpaz M, Estrov Z, O'Brien S, Kurzrock R, Kantarjian HM.The biology of chronic myeloid leukemia. N Engl J Med. 1999; 341(3): 164-172

3. Clarkson B, Strife A, Wisniewski D, Lambek CL, Liu C.Chronic myelogenous leukemia as a paradigm of early cancer and possible curative strategies. Leukemia. 2003; 17(7): 1211-1162.

4. Goldman JM, Melo JV.Chronic myeloid leukemia--advances in biology and new approaches to treatment. N Engl J Med. 2003; 349(15): 1451-1464.

5. Ben-Neriah Y, Daley GQ, Mes-Masson AM, Witte ON, Baltimore D. The chronic myelogenous leukemia-specif-ic P210 protein is the product of the bcr/abl hybrid gene. Science. 1986; 233(4760): 212-214.

6. Calabretta B, Perrotti D.The biology of CML blast cri-sis. Blood. 2004; 103(11): 4010-4022.

7. Shepherd P, Suffolk R, Halsey J, Allan N. Analysis of molecular breakpoint and m-RNA transcripts in a pro-spective randomized trial of interferon in chronic myeloid leukaemia: no correlation with clinical features, cytoge-netic response, duration of chronic phase, or survival. Br J Haematol. 1995; 89(3): 546-554.

8. Melo JV. The diversity of BCR-ABL fusion proteins and their relationship to leukemia phenotype. Blood. 1996; 88: 2375-2384.

9. Li S, Ilaria Jr RL, Million RP, Daley GQ, Van Etten RA. The P190, P210, and P230 forms of the BCR/ABL onco-gene induce a similar chronic myeloid leukemia-like syn-drome in mice but have different lymphoid leukemogenic activity. J Exp Med. 1999; 189(9): 1399-1412.

10. Wang JY.Abl tyrosine kinase in signal transduction and cell-cycle regulation. Curr Opin Genet Dev. 1993; 3(1): 35-43.

11 Kurzrock R, Gutterman JU, Taplaz M. The molecu-lar genetics of Philadelphia positive leukemias. N Engl J Med. 1988; 319(15): 990-988.

12. Pigneux A, Mahon FX, Jazwiec B, Pigeonnier V, Ripoche J, Reiffers J. Effect of antisense oligonucle-otides on CD34+ cells from chronic myeloid leukemia. Leuk Lymphoma. 2000; 36(5-6): 569-578.

13. Vester B, Wengel J. LNA (locked nucleic acid): high-affinity targeting of complementary RNA and DNA. Bio-chemistry. 2004; 43(42): 13233-13241.

14. McTigue PM, Peterson RJ, Kahn JD.

Sequence-de-pendent thermodynamic parameters for locked nucleic acid (LNA)-DNA duplex formation. Biochemistry. 2004; 43(18): 5388-5405.

15. Summerton J, Weller D. Morpholino antisense oli-gomers: design, preparation, and properties. Antisense Nucleic Acid Drug Dev. 1997; 7(3): 187-195.

16. Summerton J. Morpholino antisence oligos: appli-cations in biopharmaceutical research. IPT. 2005; 17: 32-37.

17. Klee EW, Shim KJ, Pickart MA, Ekker SC, Ellis LB. AMOD: a morpholino oligonucleotide selection tool. Nucleic Acids Res. 2005;33(Web Server issue): W506-511.

18. Apperley J. Mechanisms of resistance to imatinib in chronic myeloid luakemia. Lancet oncol. 2007; 8: 1018-1029.

19. Wilda M, Fuchs U, Wossmann W, Borkhardt A. Kill-ing of leukemic cells with a BCR/ABL fusion gene by RNA interference (RNAi). Oncogene. 2002(37); 21: 5716-5724.

20. Scherr M, Battmer K, Winkler T, Heidenreich O, Ganser A, Eder M. Specific inhibition of bcr-abl gene expression by small interfering RNA. Blood. 2003(4); 101: 1566-1569.

21.Withey JM, Harvey AJ, Crompton MR.RNA interfer-ence targeting of Bcr-Abl increases chronic myeloid leukemia cell killing by 17-allylamino-17-demethoxy-geldanamycin. Leuk Res. 2006; 30(5): 553-560.

22. Koldehoff M, Steckel NK, Beelen DW, Elmaagacli AH. Synthetic small interfering RNAs reduce bcr-abl gene expression in leukaemic cells of de novo Philadel-phia(+) acute myeloid leukaemia. Clin Exp Med. 2006; 6(1): 45-57.

23. Summerton J. Morpholino antisence oligomers: the case for an RNase H- independent structural type. Bio-chim Biophys Acta. 1999; 1489(1): 141-158.

24. Summerton JE.Endo-Porter: a novel reagent for safe, effective delivery of substances into cells. Ann N Y Acad Sci. 2005; 1058: 62-75.

25. Collins S, Coleman H, Groudine M. Expression of bcr and bcr-abl fusion transcripts in normal and leuke-mic cells. Mol Cell Biol. 1987; 7(8): 2870-2876.

26. Melo JV, Chuah C. Resistance to imatinib mesylate in chronic myeloid leukaemia. Cancer Lett. 2007; 249(2): 121-132.

Referências

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ﻲﻟﻮﻠﺳ ﻞﺧﺍﺩ ﻞﻜﺷ ﻱﻭﺮﻛ ﻰﻳﺎﺸﻏ ﻚﺗ ﻱﺎﻫ ﻚﻣﺍﺪﻧﺍ ﺎﻫ ﻡﻭﺰﻴﺴﻛﺍﺮﭘ ﻡﺎﻤﺗﺭﺩ ﻭ ﺪﻨﺷﺎﺑ ﻲﻣ ﺮﺘﻣﻭﺮﻜﻴﻣ ﻢﻫﺩ ﻚﻳ ﻰﻟﺍ ﻚﻳ ﺮﻄﻗ ﻪﺑ ﻪﻛ ﺪﻨﺘﺴﻫ ﻪﭼ ﺮﮔﺍ .(1) ﺪﻧﺭﺍﺩ ﺩﻮﺟﻭ ،ﻥﺎﺴﻧﺍ ﺎﺗ ﺮﻤﺨﻣﺯﺍ ،ﻲﺗﻮﻳﺭﺎﻛﻮﻳ ﻱﺎﻫ

ﮏﯾ ناﻮﻨﻋ ﻪﺑ نﻮﻣزآ هوﺮﮔ ﺎﺑ ﺮﮕﻧﻮﻣزآ ﯽﻫاﺮﻤﻫ و رﻮﻀﺣ ﺮﯿﺛﺎﺗ ﺎﺗ ﺪﻨﺘﻓﺮﮔ نآ شﺰﯿﮕﻧا ﺮﺑ ﺮﺛﻮﻣ ﻞﻣﺎﻋ ﺎﺑ

ﻲﻣ ﻥﺎﺸﻧ ﺩﻮﺧ ﺯﺍ ﻱﺮﺘﺸﻴﺑ ﺕﺍﺮﻴﻴﻐﺗ ﻂﻳﺍﺮﺷ ﺭﺩ ﻩﺪﺸﻧ ﻦﻴﻴﻌﺗ ﺶﻴﭘ ﺯﺍ ﻱﺎﻬﺘﻟﺎﺣ ﺭﺩ ﺯﻭﺩ ﻊﻳﺯﻮﺗ ﺶﻳﺎﻤﻧ ﺖﻬﺟ ﻪﻣﺎﻧﺮﺑ ﻚﻳ ﻪﺑ ﺯﺎﻴﻧ ،ﺪﻫﺩ.

ﺭﺎﺘﺧﺎﺳ ﻦﻴﻟﻭﺍ ﺎﺒﻳﺮﻘﺗ ﻭ ﺪﻨﺘﺴﻫ ﻁﺎﺒﺗﺭﺍ ﺭﺩ ﻰﻟﻮﻠﺳ ﻥﻭﺮﺑ ﻪﻨﻴﻣﺯ ﻩﺩﺎﻣ ﺎﺑ ﺢﻄﺳ ﻰﻣ ﺮﻈﻧ ﻪﺑ ﻭ ﺪﻧﺭﺍﺩ ﺶﻨﻛ ﻢﻫﺮﺑ ﺎﻬﻧﺁ ﺎﺑ ﻯﺩﺎﻴﻨﺑ ﻯﺎﻫ ﻝﻮﻠﺳ ﻪﻛ ﺖﺳﺍ ﻰﻟﻮﻜﻟﻮﻣ. .(16) ﺪﺷﺎﺑ ﺖﻴﻤﻫﺍ ﺰﺋﺎﺣ ﺎﻬﻧﺁ ﺰﻳﺎﻤﺗ

ﺖﻳﻮﻘﺗ ﺍﺭ ﻱﺭﻮﻣﻮﺗ ﺪﺿ ﻱﺎﻫ ﺦﺳﺎﭘ ﻥﺁ ﻝﺎﺒﻧﺩ ﻪﺑ ﻭ ﻲﻨﻤﻳﺍ ﻱﺎﻫ ﺦﺳﺎﭘ ﺖﺳﺍ ﺕﺭﻮﺻ ﻪﺑ ﻦﻴﻟﺎﻔﻜﻧﺍ - ﺖﻣ ﺮﺛﺍ ﻲﻠﺒﻗ ﺕﺎﻌﻟﺎﻄﻣ ﺭﺩ ﻪﻜﻨﻳﺍ

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