284
ﻒﺠﻧ يﺮﻔﻌﺟ ﻦﺴﺣ ﺪﻤﺤﻣ يدﺎﺑآ
نارﺎﮑﻤﻫ و
ﺳرﺮﺑ ﯽ ﻟﯿ ﻠﭘﻮﭙ ﯿ ﺎﻫﺮﻤ ي وﺎﺣ ي ﻟﻮﻧﺎﻧ ﯿ موزﻮﭙ ﻠﭘ و ﯽ ﺗا ﯿ ﻦﻠ ا ﯾ ﻤﯿ ﻦ اﺮﺑ ي ﻪﺑ نژ لﺎﻘﺘﻧا
ﻟﻮﻠﺳ هدر ﯽ
تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ PC3
ﺮﻔﻌﺟ ﻦﺴﺣ ﺪﻤﺤﻣ ي
ﻒﺠﻧ دﺎﺑآ ي
1
،Ϯ
ﻌﺳ ﯿ هﺪ ﺮﮑﺴﻋ ﯾ نﺎ
2 و 3
ﻤﻇﺎﮐ ﺎﺿر ،Ϯ
ﯽ
ﻮﮑﺳا ﯽﯾ ﺑ ،1
ﯿ نﮋ ﻧ ﻪﮑﺋﻼﻣ ﯿ ﻮﮑ ﯾ ﯽ ﺪﻬﻣ ،4
ي
ﺎﺿر ﯽﯾ ﺳ ،1
ﯿﺪ ﻤﺣ ﯿ ﺪ ﺎﻗآ ﯽﯾ ﺘﺨﺑ ﯿ رﺎ ي
1 و 5
*
1 - ز هوﺮﮔ ﯾ ﺖﺴ وﺎﻨﻓ ر ي يروﺎﻨﻓﻮﻧﺎﻧ و ﮑﺷﺰﭘ ﯽ
،
ﮑﺷﺰﭘ هﺪﮑﺸﻧاد ﯽ
، ﮑﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ﯽ
،ﺪﻬﺸﻣ
ا ،ﺪﻬﺸﻣ ﯾ ناﺮ .
2 - هوﺮﮔ ﺑﯿ ژﻮﻟﻮﻨﮑﺗﻮ ي ﮑﺷﺰﭘ ﯽ
، هﺪﮑﺸﻧاد
ﭘﯿ ﮑﺷﺰﭘاﺮ ﯽ
، ﮑﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ﯽ
ﺖﺑﺮﺗ
ﺣ ﯿ رﺪ ﯾ
،ﻪ ﺣ ﺖﺑﺮﺗ ﯿ رﺪ ﯾ
،ﻪ اﯾ ناﺮ .
3 - ﻘﺤﺗ ﺰﮐﺮﻣ ﯿ تﺎﻘ مﻮﻠﻋ هﺎﮕﺸﻧاد ،بﺎﺼﻋا مﻮﻠﻋ
ﮑﺷﺰﭘ ﯽ ﺣ ﺖﺑﺮﺗ ﯿ رﺪ ﯾ
،ﻪ ﺣ ﺖﺑﺮﺗ ﯿ رﺪ ﯾ
،ﻪ اﯾ ناﺮ .
4 - ﻘﺤﺗ ﺰﮐﺮﻣ ﯿ تﺎﻘ روﺎﻨﻓﻮﻧﺎﻧ ي
، روﺎﻨﻓ هﺪﮑﺸﻫوﮋﭘ ي
وراد ﯽﯾ
، ﮑﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ﯽ
ا ،ﺪﻬﺸﻣ ،ﺪﻬﺸﻣ ﯾ ناﺮ .
5 - ﻪﺘﺴﻫ ﻘﺤﺗ ﯿ تﺎﻘ ﺑﯿ ﺗﺎﻣرﻮﻔﻧاﻮ ﯿ
،ﮏ ﻮﻠﻋ هﺎﮕﺸﻧاد م
ﮑﺷﺰﭘ ﯽ ا ،ﺪﻬﺸﻣ ،ﺪﻬﺸﻣ ﯾ
ناﺮ .
ﺮﻔﻌﺟ ﻦﺴﺣ ﺪﻤﺤﻣϮ
ي ﻒﺠﻧ دﺎﺑآ ي ﻌﺳ و ﯿ هﺪ
ﺮﮑﺴﻋ ﯾ نﺎ ﻪﺑ ﻮﻧ كﺮﺘﺸﻣ ترﻮﺻ ﯾ
هﺪﻨﺴ .ﺪﻨﺘﺴﻫ لوا
:لﻮﺌﺴﻣ هﺪﻨﺴﯾﻮﻧ*
،ﺪﻬﺸﻣ ﻣ ﯿ ناﺪ دازآ ي
، مﻮﻠﻋ هﺎﮕﺸﻧاد
ﮑﺷﺰﭘ ﯽ
، ﮑﺷﺰﭘ هﺪﮑﺸﻧاد ﯽ
، ز هوﺮﮔ ،مﻮﺳ ﻪﻘﺒﻃ ﯾ
ﺖﺴ
روﺎﻨﻓ ي ﯽﮑﺷﺰﭘ . ﻦﻔﻠﺗ : 38002300 - 051
E-mail: [email protected]
فﺪﻫ و ﻪﻨﯿﻣز ﻞﻣﺎﺣﻮﻧﺎﻧ:
ﺎﻫ ي ﻏ ﯿﺮ وﯾ ﺳوﺮ ﯽ ﻟ ﻞــﺜﻣ ﯿ موزﻮﭙ ﻠﭘ و ﺎــﻫ ﯿ ﺮﻤ ﺎــﻫ ي ﺗﺎــﮐ ﯿ ﻧﻮ ﯽ اراد ي ﻠﺑﺎﻗ ــﯿ ﺖ زرﻮﺘــﺳد ي ــﺴﺑ ﯿ رﺎ ي ﻪــﺑ
لﻮﻠﺳ ﻪﺑ نژ لﺎﻘﺘﻧا رﻮﻈﻨﻣ ا ﺎﺑ .ﺪﻨﺘﺴﻫ ﺎﻫ
ﯾ ﻦ ارﺎﮐ دﻮﺟو ﯽﯾ نآ لﺎﻘﺘﻧا و زا ﺮــﺘﻤﮐ ﺎﻫ ﯾ
سوﺮ زا فﺪــﻫ .ﺖــﺳا ﺎــﻫ ا
ــﯾ ﻦ ــﻌﻟﺎﻄﻣ ،ﻪ
ﮐﺮﺗ ﯿ ﺐ ﻟﯿ موزﻮﭙ ﻠﭘ و ﺎﻫ ﯿ ﺮﻤ ﺎﻫ ي ﺗﺎﮐ ﯿ ﻧﻮ ﯽ اﺮﺑ ي ﻬﺗ ﯿﻪ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ و هﺮﻬﺑ رادﺮﺑ ي ــﺻﻮﺼﺧ زا ﯿ تﺎ ﻟ بﻮــﻠﻄﻣ ﯿ زﻮــﭙ مو ﻠﭘ و ﯿ ﺮــﻤ
ﺑﻪ ﻢﻫ ترﻮﺻ ﻣ نﺎﻣز ﯽ .ﺪﺷﺎﺑ
ﯽﺳرﺮﺑ شور اﯾ:
ﻦ ﻪﻠﺧاﺪﻣ ﻪﻌﻟﺎﻄﻣ ا
ي - د رد ﯽﺑﺮﺠﺗ ﮑﺷﺰﭘ هﺪﮑﺸﻧا ﯽ
ﮑــﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ﯽ
دروﺮــﻓ زا ﺪﻬــﺸﻣ ﯾ
ﻦ ﺎــﺗ ﺑ ﻦــﻤﻬ
1396 ا رد .ﺖﻓﺮﮔ مﺎﺠﻧا ﯾ
ﻦ ،ﻪﻌﻟﺎﻄﻣ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﺎﻫ ﯽﯾ ﺎﭘ ﺮﺑ ﯾﻪ ﻠﭘ)PEI
ﯽ ﺗاﯿ ﻦﻠ اﯾ ﻤﯿ ﻦ ( ﺎــﺑ و ﻟﻮــﮑﻟﻮﻣ نزو ﯽ
25 و 10 ﮐ ﯿ ﻮــﻠ
ﻟ ﺖﺒﺴﻧ و نﻮﺘﻟاد ﯿ
موزﻮﭙ ﻠﭘ ﻪﺑ ﯿ ﺮﻤ 1 ﻪﺑ 1 ﻼﭘ زا هدﺎﻔﺘــﺳا ﺎﺑ ﻤــﺳ
ﯿ ﺪ وﺎــﺣ ي ﺮﮕﻧﺎــﺸﻧ ﺗوﺮــﭘ)GFP
ﯿﯿ ﻦ ــﺳرﻮﻠﻓ ﺘﻨ ﯽ ﻬﺗ (ﺰﺒــﺳ ــﯿ ﻪ
دﺮﮔ ﯾﺪ . ﯽﮔﮋﯾو يﺎﻫ ﻓﯿ ﺰﯾ ﮑ ﯽ و ﺷ ﯿﻤ ﯿﺎ ﯽﯾ ﻞﻣﺎﺣ ﺎﻫ ي هﺪﺷ ﺰﺘﻨﺳ ﺪﻨﻧﺎﻣ ﻤــﺳ ،هزاﺪﻧا ﯿ ﺖ ﻟﻮﻠــﺳ ﯽ ﻐﺘﻣ) ﯿﺮ ﺎــﻫ ي ﻠــﺻا ﯽ و ( طﺎــﺒﺗرا
نآ ﻠﺑﺎﻗ ﺎﺑ ﺎﻫ ﯿ ﺖ لﻮﻠﺳ رد نژ لﺎﻘﺘﻧا ﺎﻫ
ي ﻧﺎﺴﻧا تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ ﯽ
(PC3)
زرا درﻮﻣ ﯾﺑﺎ ﯽ .ﺖﻓﺮﮔ راﺮﻗ
ﻪﺘﻓﺎﯾ :ﺎﻫ ﻣﺎﻤﺗ ﯽ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ رد ﺎﻫ 4 /0 ﺎــﺑC/P=
ﺎﭘ ــﯾ ﻪ ود ﺮــﻫ ﯽﻟﻮــﮑﻟﻮﻣ نزو ــﻠﭘ)PEI
ﯽ ﺗاﯿ ﻦﻠ اﯾ ــﻤ ﯿ ﻦ ﻪــﺑ ردﺎــﻗ ( اﺰــﻓا ﯾ ﺶ
ﻞﻣﺎﺣ ﻪﺑ ﺖﺒﺴﻧ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺎﻫ
ي ﺎﭘ ﯾﻪ ﻠﭘ) ﯽ ﺗاﯿ ﻦﻠ اﯾ ﻤﯿ ﻦ ﻟ و ﯿ موزﻮﭙ دﻮﺑ ( ) 05 /0
P<
، 001 /0
P<
( ﻟﺎﺣرد ، ﯽ ﻤــﺳ ﻪــﮐ ﯿ ﺖ ﻟﻮﻠــﺳ ﯽ
ﺮﺘﻤﮐ ي ﻧﯿ ﺰ ﻟ ﻪﺑ ﺖﺒﺴﻧ ﯿ موزﻮﭙ زرو ﺖﺳد ي ﻣ نﺎﺸﻧ هﺪﺸﻧ ﯽ
.داد
ﻪﺠﯿﺘﻧ :يﺮﯿﮔ ﺎﺘﻧ ﯾ ﺞ اﯾ ﻦ ﻣ نﺎﺸﻧ ﻪﻌﻟﺎﻄﻣ ﯽ
ﺪﻫد ﭙﯿﻟ ﯽﻠﭘﻮ ﺲﮑﻠﭘ ﮐﺮﺗ زا ﻞﺻﺎﺣ ﯿ
ﺐ ــﻠﭘ)PEI
ﯽ ﺗاﯿ ﻦﻠ اﯾ ــﻤ ﯿ ﻦ ﻟ و ( ﯿ زﻮــﭙ مو ﻣ ﯽ ﺪــﻧاﻮﺗ
دﺎﻘﻣ رد ﯾﺮ ﻪﺑ ﻢﮐ ﺪﻣآرﺎﮐ ترﻮﺻ ي
ﺎﻤﻧ ﻞﻘﺘﻨﻣ لﻮﻠﺳ ﻪﺑ ار نژ ﯾﺪ
ﻟﺎﺣرد ﯽ ﻤﺳ ﻪﮐ ﯿ ﺖ ﻟﻮﻠﺳ ﻢﮐ ﯽ ﺒــﺳﺎﻨﻣ هزاﺪــﻧا و ﯽ
ﻘﻣ رد ــﯿ سﺎ
ﻮﻧﺎﻧ دراد .
ﮐ تﺎﻤﻠ ﮐ يﺪﯿﻠ : ﮏﯿﻨﮑﺗ ،ﯽﻧژ لﺎﻘﺘﻧا يﺎﻫ موزﻮﭙﯿﻟ
،ﺎﻫ لﻮﻠﺳ يﺎﻫ ،PC-3
ﯽﻧژ لﺎﻘﺘﻧا .
نژ ﻒﻠﺘﺨﻣ يﺎﻫشور عاﺪﺑا زا لﺎﺳ ﻦﯾﺪﻨﭼ ﺖﺷﺬﮔ دﻮﺟو ﺎﺑ داﻮﻣ ﺎﯾ ﺎﻫﺪﯿﺗﻮﺌﻠﮐﻮﻧﻮﮕﯿﻟوا ﺪﻣآرﺎﮐ لﺎﻘﺘﻧا ،نﺎﺴﻧا مﻮﻧژ هژوﺮﭘ و ﯽﻧﺎﻣرد
لﻮﻠﺳ ﻪﺑ ﯽﮑﯿﺘﻧژ ﺮﻓ رد ﺶﻟﺎﭼ ﻦﯾﺮﺘﻤﻬﻣ زﻮﻨﻫ ﺎﻫ
آ ﯽﻧﺎﻣرد نژ ﺪﻨﯾ
يرﺎﻤﯿﺑ نﺎﻃﺮﺳ ﻪﻠﻤﺟ زا يرﺎﯿﺴﺑ يﺎﻫ ﺖﺳا ﺎﻫ
. ﻞﻣﺎﺣ1
ﻘﺘﻧا يﺎﻫ ﻪﺑ ﯽﻧژ لﺎ
ﻞﻣﺎﺣ ﯽﻠﺻا هوﺮﮔ ود لﺎﻘﺘﻧا ياﺮﺑ ﯽﺳوﺮﯾوﺮﯿﻏ و ﯽﺳوﺮﯾو يﺎﻫ
DNA
،يﺪﯿﻤﺳﻼﭘ
siRNA
، وmRNA miRNA
ﯽﻣ ﻢﯿﺴﻘﺗ ﻢﺘﺴﯿﺳ .ﺪﻧﻮﺷ يﺎﻫ
(ﯽﻧژ لﺎﻘﺘﻧا) ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺖﯿﻠﺑﺎﻗ و ﺪﻧراد ﯽﯾﻻﺎﺑ ﯽﯾآرﺎﮐ ﯽﺳوﺮﯾو هداد نﺎﺸﻧ دﻮﺧ زا ﯽﺑﻮﺧ ﺎﺑ ﯽﻨﯿﻟﺎﺑ ﯽﯾﺎﻣزآرﺎﮐ ﻪﻌﻟﺎﻄﻣ ﻦﯾﺪﻨﭼ و ﺪﻧا
ﺣ زا هدﺎﻔﺘﺳا ﻞﻣﺎ
ﺖﺳا ﯽﺳرﺮﺑ لﺎﺣ رد ﯽﺳوﺮﯾو يﺎﻫ .
2 و
ﺎﺑ3
ﻦﯾا لﺎﺣ
يدراﻮﻣ ﻦﯿﯾﺎﭘ ﺖﯿﻓﺮﻇ ﻞﻣﺎﺷ ﻞﻣﺎﺣ
ﯽﺳوﺮﯾو يﺎﻫ لﺎﻘﺘﻧا ﺖﯿﻠﺑﺎﻗ)
ﻪﻣﺪﻘﻣ
ﻪﻟﺎﻘﻣ ﺻا ﻞﯿ
292 ﺎﺗ284 يﺎﻫﻪﺤﻔﺻ ،5 هرﺎﻤﺷ، 78 هرود ،1399دادﺮﻣ ،ناﺮﻬﺗ ﯽﮑﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد ،ﯽﮑﺷﺰﭘ هﺪﮑﺸﻧاد ﻪﻠﺠﻣ
هﺪﯿﮑﭼ
:ﺖﻓﺎﯾرد
02 / 12 / 1398 ﺶﯾاﺮﯾو : 09 / 12 / 1398 :شﺮﯾﺬﭘ 23 / 05 / 1399 :ﻦﯾﻼﻧآ 30 / 05 /
1399
Downloaded from tumj.tums.ac.ir at 13:01 IRST on Friday December 18th 2020
ﺳرﺮﺑ ﯽ ﻟﯿ ﻠﭘﻮﭙ ﯿ ﺎﻫﺮﻤ اﺮﺑ ي تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ لﻮﻠﺳ ﻪﺑ نژ لﺎﻘﺘﻧا 285
نژ لﺎﻌﻓ لﺎﻤﺘﺣا ،(ﺖﺳا يروﺮﺿ ﯽﻨﯿﻟﺎﺑ يﺎﻫدﺮﺑرﺎﮐ ياﺮﺑ گرﺰﺑ يﺎﻫ
ﺮﭘ ندﺮﮐ و نژﻮﮑﻧآ ﻮﺗ ﻪﺑ ﺎﻫ ﺦﺳﺎﭘ ﺎﻘﻟا ،مﻮﻧژ ﻪﺑ ﯽﻓدﺎﺼﺗ دورو ﻞﯿﻟد يﺎﻫ
ﻖﯾرﺰﺗ ﻪﮐ) يﻮﻗ ﯽﻨﻤﯾا ار رﺮﮑﻣ يﺎﻫ
ﯽﻣ دوﺪﺤﻣ رد تﻼﮑﺸﻣ و (ﺪﻨﮐ
نآ هﻮﺒﻧا ﺪﯿﻟﻮﺗ ﺎﻫ
ﻞﻣﺎﺣ عﻮﻧ ﻦﯾا زا هدﺎﻔﺘﺳا رد ﻞﮑﺸﻣ ﻦﯾﺮﺘﻤﻬﻣ ﺎﻫ
ﯽﻣ ﺪﺷﺎﺑ . ﻪﺑ 4
لﺎﺳ رد ﯽﻧاﻮﺟ رﺎﻤﯿﺑ لﺎﺜﻣ ناﻮﻨﻋ 1999
ﻪﺑ ﻖﯾرﺰﺗ ﻞﯿﻟد
لﺎﺳ رد ﺮﮕﯾد رﺎﻤﯿﺑ ود و ﺪﯾدﺮﮔ تﻮﻓ ﯽﺳوﺮﯾوﻮﻧدآ ﻞﻣﺎﺣ 2002
ﺎﺑ
يرﺎﻤﯿﺑ ﺪﯾﺪﺷ ﯽﻨﻤﯾا ﺺﻘﻧ
(SCID)
، ﺮﺗر ﻞﻣﺎﺣ ﻖﯾرﺰﺗ زا ﺲﭘ ﯽﺳوﺮﯾوو
ﻼﻋ ﯾ ﺪﻧداد نﺎﺸﻧ ار ﯽﻤﺳﻮﻟ زا ﯽﻤ .
5 و
6
ﻢﺘﺴﯿﺳ ﯽﻧژ لﺎﻘﺘﻧا يﺎﻫ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ
(LPP)
زا ﯽﺒﯿﮐﺮﺗ
لﻮﮑﻟﻮﻣ ﯽﻠﭘ يﺎﻫ تﻻﺎﺼﺗا ﺎﺑ ﯽﻧﻮﯿﺗﺎﮐ ﺮﻤﯿﻠﭘ و موزﻮﭙﯿﻟ ،يﺪﯿﺗﻮﺌﻠﮐﻮﻧ
ﯽﺴﻧﻻاﻮﮐﺮﯿﻏ ﺪﻨﺘﺴﻫ
ﻪﮐ ﻢﻫ ﻞﻣﺎﺷ ﺲﮑﻠﭘﻮﭙﯿﻟ بﻮﻠﻄﻣ تﺎﯿﺻﻮﺼﺧ
ﻦﯿﯾﺎﭘ ﺖﯿﻤﺳ و لﻮﺒﻗ ﻞﺑﺎﻗ ﯽﻟﻮﻠﺳ ﺖﺷادﺮﺑ ،ﻻﺎﺑ يراﺪﯾﺎﭘ و
ﻢﻫ
ﯽﮔﮋﯾو ﯽﻠﭘ ﺐﺳﺎﻨﻣ يﺎﻫ ﺰﯾﺎﺳ ،ﻻﺎﺑ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺖﯿﻠﺑﺎﻗ ﻪﻠﻤﺟ زا ﺲﮑﻠﭘ
ار لﻮﻠﺳ ﻪﺑ ﯽﮑﯿﺘﻧژ هدﺎﻣ لﺎﻘﺘﻧا ياﺮﺑ موزوﺪﻧآ زا راﺮﻓ ﺖﯿﻠﺑﺎﻗ و ﮏﭼﻮﮐ ﯽﻣ اراد ﺷﺎﺑ ﻨ ﺪ.
ﯽﻠﭘﻮﭙﯿﻟ7
ﺲﮑﻠﭘ نوﺰﻓا ﻪﮐ ﯽﯾﺎﯾاﺰﻣ ﺮﺑ نﺎﯿﺑ
ﻪﺑ ﺪﺷ اراد ﻞﯿﻟد
هدﺮﺘﺴﮔ ﺢﻄﺳ ندﻮﺑ
، هوﺮﮔ لﺎﺼﺗا ﺖﯿﻠﺑﺎﻗ ر يرﺎﯿﺴﺑ يدﺮﮑﻠﻤﻋ يﺎﻫ
ﻪﺑ ا
ﻪﺑ و دراد يﺮﻤﯿﻠﭘ ﺎﯾ ﯽﻣوزﻮﭙﯿﻟ ﺪﺣاو يزرﻮﺘﺳد ﻞﺑﺎﻗ ﺪﺣاو ﮏﯾ ناﻮﻨﻋ
،ﺮﻤﯿﻠﭘ عﻮﻧ و موزﻮﭙﯿﻟ ﺐﯿﮐﺮﺗ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﻦﯾاﺮﺑﺎﻨﺑ .ﺖﺳا هﺪﺷ حﺮﻄﻣ لﺪﻣ ﯽﻠﭘﻮﭙﯿﻟ زا ﯽﻔﻠﺘﺨﻣ يﺎﻫ ﻪﺘﺧﺎﺳ ﯽﮑﯿﺘﻧژ هدﺎﻣ لﺎﻘﺘﻧا ياﺮﺑ ﺲﮑﻠﭘ
ﺖﺳا هﺪﺷ . ﻪﺑ ﯽﻠﭘﻮﭙﯿﻟ لﺎﺜﻣ ناﻮﻨﻋ هﺪﺷ ﻞﺼﺘﻣ نوزﺎﺘﻣاﺰﮔد يوﺎﺣ ﺲﮑﻠﭘ
ﯽﻠﭘ ﻪﺑ لﻮﻠﺳ ﻪﺑ ﯽﻧژ لﺎﻘﺘﻧا ياﺮﺑ ﻦﯿﻤﯾا ﻦﻠﯿﺗا يﺎﻣﻮﺘﺳﻼﺑورﻮﻧ يﺎﻫ
ﯽﺷﻮﻣ
A 2 Neuro
ﺖﺳا هﺪﺷ هدﺎﻔﺘﺳا .
8
ﯽﻠﭘﻮﭙﯿﻟ رﺎﺘﺧﺎﺳ رد هﺪﺷ ﻪﻌﻟﺎﻄﻣ ﺮﻤﯿﻠﭘ ﻦﯾﺮﺘﺠﯾار ﺲﮑﻠﭘ
ﯽﻠﭘ ،ﺎﻫ ﻦﻠﯿﺗا
ﻦﯿﻤﯾا
(PEI)
ﯽﻟﻮﮑﻟﻮﻣ نزو ﻪﺑ ﺮﻤﯿﻠﭘ ﻦﯾا ﯽﻧژ لﺎﻘﺘﻧا ﺖﯿﻠﺑﺎﻗ ﻪﮐ ﺖﺳا
.دراد ﯽﮕﺘﺴﺑ نآ ﺖﯿﻠﺑﺎﻗ ﻻﺎﺑ ﯽﻟﻮﮑﻟﻮﻣ نزو ﺎﺑPEI
ﯽﺑﻮﺧ ﻦﺸﮑﻔﺴﻧاﺮﺗ
ﯽﻣ نﺎﺸﻧ ﺰﯿﻧ يرﺎﯿﺴﺑ ﺖﯿﻤﺳ ﺎﻣا ﺪﻧراد ﺎﺑ يرﺎﯿﺴﺑ تﺎﻌﻟﺎﻄﻣ ﻦﯾاﺮﺑﺎﻨﺑ ،ﺪﻫد
هاﺮﻤﻫ ﯽﻟﻮﻠﺳ ﺖﯿﻤﺳ ﺶﻫﺎﮐ ياﺮﺑ ﺮﻤﯿﻠﭘ ﻦﯾا ﻪﺑ ﻒﻠﺘﺨﻣ ﻞﻣاﻮﻋ لﺎﺼﺗا ﻆﻔﺣ ﺎﺑ .ﺖﺳا ﻪﺘﻓﺮﮔ ترﻮﺻ ﯽﻧﺎﺳر نژ ﺖﯿﻠﺑﺎﻗ ﯽﺧﺮﺑ ﺮﮕﯾد فﺮﻃ زا
زا هدﺎﻔﺘﺳا ﻪﺑ تﺎﻌﻟﺎﻄﻣ ﻦﯿﯾﺎﭘ ﯽﻟﻮﮑﻟﻮﻣ نزو ﺎﺑ ﯽﯾﺎﻫPEI
ﺗ ﺎﯾ و ﺮﺗ ﺐﯿﮐﺮ
نآ موزﻮﭙﯿﻟ ﺎﺑ ﺎﻫ ﻪﺘﺧادﺮﭘ ﺎﻫ
.ﺪﻧا ﻦﯾا زا 9
ﺮﯾدﺎﻘﻣ ﺶﻫﺎﮐ ﺎﺑ ور رد PEI
ﯽﻠﭘﻮﭙﯿﻟ رﺎﺘﺧﺎﺳ ﯽﻣ رﺎﻈﺘﻧا ﺰﯿﻧ يﺮﺘﻤﮐ ﺖﯿﻤﺳ ،ﺲﮑﻠﭘ
دور .
10 و
فﺪﻫ11
زا
ا ﻦﯾ
،ﻪﻌﻟﺎﻄﻣ ﺐﯿﮐﺮﺗ ﻠﭘ ﯽ ﺗا ﻦﻠﯿ ا ﻦﯿﻤﯾ ﺎﻫ ي ﺎﺑ ود ﻟﻮﮑﻟﻮﻣ نزو ﯽ
توﺎﻔﺘﻣ
) 25 و 10 ﮐ ﻮﻠﯿ ﻟ و (نﻮﺘﻟاد موزﻮﭙﯿ
وﺎﺣ ي لوﺮﺘﺴﻠﮐ ﻪﺑ ﻬﺗ رﻮﻈﻨﻣ ﻪﯿ
ﻟ ﯽﻠﭘﻮﭙﯿ ﻘﻣ رد هزاﺪﻧا ﺎﺑ ﺲﮑﻠﭘ سﺎﯿ
ﻮﻧﺎﻧ .دﻮﺑ
ﻪﻠﺧاﺪﻣ ﻪﻌﻟﺎﻄﻣ ﻦﯾا يا
- ﯽﺑﺮﺠﺗ مﻮﻠﻋ هﺎﮕﺸﻧاد ﯽﮑﺷﺰﭘ هﺪﮑﺸﻧاد رد
ﻦﻤﻬﺑ ﺎﺗ ﻦﯾدروﺮﻓ زا ﺪﻬﺸﻣ ﯽﮑﺷﺰﭘ 1396
ﺖﻓﺮﮔ مﺎﺠﻧا درﻮﻣ داﻮﻣ .
ﯽﻠﭘ ،ﻞﻣﺎﺷ ﻪﻌﻟﺎﻄﻣ ﻦﯾا رد هدﺎﻔﺘﺳا ﻪﺧﺎﺷ ﻦﯿﻤﯾا ﻦﻠﯿﺗا
ﯽﻟﻮﮑﻟﻮﻣ نزو) راد
25 ﯿﮐ ﯽﻠﭘ ،(نﻮﺘﻟادﻮﻠ ﻪﺧﺎﺷ ﻦﯿﻤﯾا ﻦﻠﯿﺗا
ﯽﻟﻮﮑﻟﻮﻣ نزو) راد 10
،نﻮﺘﻟادﻮﻠﯿﮐ
(Polysciences, Inc., Pennsylvania, USA)
، يد ﻞﯿﺘﻣ
ﺪﯿﺴﮐﻮﻔﻟﻮﺳ
(DMSO)
فﺮﻌﻣ ، 3 - ) 4 و 5 - يد (ﻞﯿﻟوزاﺮﺘﺗ ﻞﯿﺘﻣ -
2 و 5 –
يد ﺎﯾ مﻮﯿﻟوزاﺮﺘﺗ ﻞﯿﻨﻓ ﺎﻤﮕﯿﺳ ﺖﮐﺮﺷ) MTT
،(ﺎﮑﯾﺮﻣآ ،ﭻﯾرﺪﻟآ
ﯽﻧﻮﯿﺗﺎﮐ ﺪﯿﭙﯿﻟﻮﻔﺴﻓ و لوﺮﺘﺴﻠﮐ 1
و 2 - يد ﻞﯿﺌﻟوا - 3 - مﻮﯿﻧﻮﻣآ ﻞﯿﺘﻤﯾﺮﺗ
نﺎﭘوﺮﭘ
(DOTAP)
، ﻂﯿﺤﻣ ﺖﺸﮐ مﺮﺳ ،DMEM
ﻦﯿﻨﺟ يوﺎﮔ ،FBS
ﯽﻨﭘ
،ﻦﯿﻠﯿﺳ
،ﻦﯿﺳﺎﻣﻮﺘﭘﺮﺘﺳا
،ﻦﯿﻣﺎﺗﻮﻠﮔ ﻦﯿﺴﭙﯾﺮﺗ
(Gibco TM,
Germany)
ﺪﯿﻤﺳﻼﭘ ،
pEGFPN1
)
Promega Corporation,
Wisconsin, USA
لﻮﻠﺳ هدر و ( وPC3
HEK-293
زا ﻮﺘﯿﺘﺴﻧا رﻮﺘﺳﺎﭘ
ناﺮﯾا ﯽﻣ ﺪﺷﺎﺑ .
شور مووزﻮﭙﯿﻟ ﺪﻟﻮﺗ ياﺮﺑ هدﺎﻔﺘﺳا درﻮﻣ لﻼﺣ ﺮﯿﺨﺒﺗ شور ﺎﻫ
(Solvent evaporation)
ﺪﯿﭙﯿﻟﻮﻔﺴﻓ .دﻮﺑ
DOTAP
ﺎﺑ لوﺮﺘﺴﻠﮐ و
ﯽﻟﻮﻣ ﺖﺒﺴﻧ 1
ﻪﺑ 1 :مرﻮﻓوﺮﻠﮐ لﻼﺣ رد لﻮﻧﺎﺘﻣ
ﺖﺒﺴﻧ ﺎﺑ) 2 ﻪﺑ 1 ﻞﺣ (
.ﺪﯾدﺮﮔ ﺲﭘ راّود هﺪﻨﻨﮐ ﺮﯿﺨﺒﺗ هﺎﮕﺘﺳد زا هدﺎﻔﺘﺳا ﺎﺑ ﯽﻟآ لﻼﺣ ﺮﯿﺨﺒﺗ زا
ﯿﻟ كزﺎﻧ ﻪﯾﻻ (يرﺎﺗور) ﯽﻫﺪﺑآ هﺰﯿﻧﻮﯾد بآ زا هدﺎﻔﺘﺳا ﺎﺑ هﺪﺷ ﻞﯿﮑﺸﺗ ﺪﯿﭙ
موزﻮﭙﯿﻟ .ﺪﺷ تﺪﻣ ﻪﺑ ﻪﺘﻓﺮﮔ ﻞﮑﺷ يﺎﻫ
ود ﻂﯿﺤﻣ يﺎﻣد رد ﺖﻋﺎﺳ
هزاﺪﻧا ﺶﻫﺎﮐ ياﺮﺑ و ﺪﺷ يراﺪﻬﮕﻧ
، موزﻮﭙﯿﻟ زا هﺪﺷ ﻪﺘﺧﺎﺳ يﺎﻫ
هزاﺪﻧا ﺎﺑ ﻪﺗارﺪﯿﻫﻮﺑﺮﮐ يﺎﻫﺮﺘﻠﯿﻓ ي
1 ، 8/
0 ، 4/
0 و 1µm
/0 هداد رﻮﺒﻋ
.ﺪﺷ ﯽﻠﭘ زا يوﺎﺴﻣ ﻢﺠﺣ ﻈﻠﻏ) ﻦﯿﻤﯾا ﻦﻠﯿﺗا
ﺖ
mg/ml
1 موزﻮﭙﯿﻟ و (
ﺖﻈﻠﻏ)
mg/ml
6 ﺎﺗ ﺪﻧﺪﺷ ﺐﯿﮐﺮﺗ ،ﻂﯿﺤﻣ يﺎﻣد رد ﺮﮕﯾﺪﮑﯾ ﺎﺑ (
ﺮﻤﯿﻠﭘﻮﭙﯿﻟ
PEI/Liposome
ﺑ ﻪ .ﺪﯾآ ﺖﺳد زا زﺎﯿﻧ درﻮﻣ ﺮﯾدﺎﻘﻣ
ﺮﻤﯿﻠﭘﻮﭙﯿﻟ ﺖﺒﺴﻧ ﺐﺴﺣ ﺮﺑ هﺪﺷ ﺰﺘﻨﺳ يﺎﻫ ﺮﻤﯿﻠﭘ ﯽﻧزو
ﺪﯿﻤﺳﻼﭘ ﻪﺑPEI
(C/P)
ﺮﯾدﺎﻘﻣ ياﺮﺑ 4/
0
،8 /0 و 2/
1 مﺮﺳ نوﺪﺑ ﯽﻟﻮﻠﺳ ﺖﺸﮐ ﻂﯿﺤﻣ رد
(pH 7.4)
ﻞﺣ ﺪﺷ رد هﺪﺷ ﻞﺣ ﺪﯿﻤﺳﻼﭘ زا زﺎﯿﻧ درﻮﻣ ﺮﯾدﺎﻘﻣ ﺎﺑ و
ﻞﯿﮑﺸﺗ ياﺮﺑ و ﺪﯾدﺮﮔ طﻮﻠﺨﻣ مﺮﺳ نوﺪﺑ ﺖﺸﮐ ﻂﯿﺤﻣ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ
تﺪﻣ ﻪﺑ 20 .ﺪﺷ ﻪﺑﻮﮑﻧا ﻂﯿﺤﻣ يﺎﻣد رد ﻪﻘﯿﻗد
ياﺮﺑ تارذ هزاﺪﻧا ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ ،ﺎﻫ ﺑ رﻮﻧ ﯽﮔﺪﻨﮐاﺮﭘ زا هدﺎﻔﺘﺳا ﺎ
ﯽﮑﯿﻣﺎﻨﯾد
Dynamic light scattering (DLS)
د رﺰﯿﻟ و ﺖﻋﺮﺳ ﺮﻠﭘا ﺞﻨﺳ
شور ﯽﺳرﺮﺑ
Downloaded from tumj.tums.ac.ir at 13:01 IRST on Friday December 18th 2020
286
ﻒﺠﻧ يﺮﻔﻌﺟ ﻦﺴﺣ ﺪﻤﺤﻣ يدﺎﺑآ
نارﺎﮑﻤﻫ و
Laser doppler velocimeter (LDV)
رد
Zetasizer - From Malvern
Panalytical, UK
ﻂﯿﺤﻣ يﺎﻣد رد و هزاﺪﻧا
ﺮﻈﻧ درﻮﻣ راﺪﻘﻣ .ﺪﺷ يﺮﯿﮔ
زا ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ يﺎﻫ رد هﺪﺷ ﺰﺘﻨﺳ 150 µl
ﺮﯿﻄﻘﺗ رﺎﺑود ﺮﻄﻘﻣ بآ
رد هﺪﺷ ﺮﺘﻠﯿﻓ ﯽﮑﯾژﻮﻟﻮﯾﺰﯿﻓ pH
(pH 7.4)
ﺎﺑ و ﺪﺷ ﻖﯿﻗر 150 µl
ﻮﻠﺤﻣ ل ﺖﻈﻠﻏ) pDNA
mg/ml
1 ياﺮﺑ و ﺪﯾدﺮﮔ طﻮﻠﺨﻣ (ﺪﯿﻤﺳﻼﭘ
ﯽﻠﭘ ﻞﯿﮑﺸﺗ تﺪﻣ ﻪﺑ ﺲﮑﻠﭘ
20 اﺮﻗ قﺎﺗا يﺎﻣد رد ﻪﻘﯿﻗد ر
.ﺪﺷ هداد
ﺲﭙﺳ 700 µl
و ﺪﺷ ﻪﻓﺎﺿا بآ تارذ هزاﺪﻧا
ﯽﺳرﺮﺑ ﺮﯾدﺎﻘﻣ .ﺪﺷ
هزاﺪﻧا رﺎﺑ ﻪﺳ زا ﺲﭘ هﺪﺷ شراﺰﮔ ﻪﺑ ،يﺮﯿﮔ
ﻦﯿﮕﻧﺎﯿﻣ ناﻮﻨﻋ
±
ارا SD
ﯾ ﻪ
.ﺖﺳا هﺪﯾدﺮﮔ
ﺪﯿﻤﺳﻼﭘ
pEGFPN1
ﺪﮐ) ﺗوﺮﭘ هﺪﻨﻨﮐ ﯿ ﻦﯿ ﺮﺗﻮﻣوﺮﭘ ﺖﺤﺗ ،GFP
يﺮﺘﮐﺎﺑ نورد (ﯽﺳوﺮﯾوﻮﻟﺎﮕﻣﻮﺘﯾﺎﺳ ﯽﻠﮐ ﺎﯿﺸﯾﺮﺷا
ﺪﯾدﺮﮔ ﻞﻘﺘﻨﻣDH5α
ﻦﯿﺴﮐﻮﺗوﺪﻧآ زا يرﺎﻋ ﺪﯿﻤﺳﻼﭘ جاﺮﺨﺘﺳا ﺖﯿﮐ زا هدﺎﻔﺘﺳا ﺎﺑ و
(QIAGEN, Germany)
.ﺪﯾدﺮﮔ جاﺮﺨﺘﺳا هﺪﻧزﺎﺳ ﺖﮐﺮﺷ رﻮﺘﺳد ﻖﺒﻃ
جﻮﻣ لﻮﻃ رد ﻪﻠﺻﺎﺣ يﺎﻫﺪﯿﻤﺳﻼﭘ صﻮﻠﺧ ﻪﺟرد و ﺖﻈﻠﻏ وA260
ﺖﺒﺴﻧ
A260/280
زا هدﺎﻔﺘﺳا ﺎﺑ ﺮﺘﻣﻮﺘﻓوﺮﺘﮑﭙﺳا
(Spectrophotometer,
Pharmaspec, Shimadzu, Japan)
ﺎﺑ يﺎﻫﺪﯿﻤﺳﻼﭘ .ﺪﯾدﺮﮔ ﯽﺳرﺮﺑ
ﺎﺑ ﺪﯿﯾﺎﺗ زا ﺲﭘ ﻞﯾﻮﮐﺮﭘﻮﺳ ﺖﻟﺎﺣ ﻦﯾﺮﺘﺸﯿﺑ رد و ﻻﺎﺑ صﻮﻠﺧ .ﺪﻧﺪﺷ بﺎﺨﺘﻧا يﺪﻌﺑ تﺎﺸﯾﺎﻣزآ ياﺮﺑ زرﻮﻓوﺮﺘﮑﻟا
لﻮﻠﺳ يﺎﻫ )PC3
(ﯽﻧﺎﺴﻧا تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ ﺎﺼﺨﺸﻣ ﺎﺑ
ت ﯽﻟﻮﻠﺳ
ATCC:CRL-1435
ﯽﻟﻮﻠﺳ هدر و
HEK-293
لﻮﻠﺳ) ﯽﻨﯿﻨﺟ ﻪﯿﻠﮐ يﺎﻫ
ﯽﻟﻮﻠﺳ تﺎﺼﺨﺸﻣ ﺎﺑ (نﺎﺴﻧا
ATCC:CRL-1573
ﺖﺸﮐ ﻂﯿﺤﻣ رد
يوﺎﺣ)DMEM
،ﺰﮐﻮﻠﮔ1g/l
2mmol
ﺎﺑ (ﻦﯿﻣﺎﺗﻮﻠﮔ 10
% يوﺎﮔ مﺮﺳ
(FBS)
لﻮﻠﺤﻣ ، ﯽﺘﻧآ ﮏﯿﺗﻮﯿﺑ يﺎﻫ ﯽﻨﭘ ﻦﯿﻠﯿﺳ 100 ﺪﺣاو) / ﯽﻠﯿﻣ و (ﺮﺘﯿﻟ
) ﻦﯿﺴﯾﺎﻣﻮﺘﭘﺮﺘﺳا
g/ml
100µ
( رد يﺎﻣد
°C
37 يوﺎﺣ رﻮﺗﺎﺑﻮﮑﻧا رد 5
%
ﺖﺸﮐCO2
هداد ﺪﻧﺪﺷ لﻮﻠﺳ.
داﺪﻌﺗ ﻪﺑ ﺎﻫ 7000
ﮏﻫﺎﭼ ﺮﻫ رد لﻮﻠﺳ
ﺖﯿﻠﭘ رد 96 و ﺪﻧﺪﺷ هداد ﺖﺸﮐ ﻪﻧﺎﺧ ﺶﯿﭘ
ﺖﺴﺗ مﺎﺠﻧا زا ﺖﯿﻤﺳ يﺎﻫ
ﻦﺸﮑﻔﺴﻧاﺮﺗ و
، تﺪﻣ ﻪﺑ 24 رﻮﺗﺎﺑﻮﮑﻧا رد ﺖﻋﺎﺳ ﯽﻟﻮﻠﺳ ﺖﺸﮐ
راﺮﻗ
.ﺪﻧﺪﺷ هداد
ﻪﺑ ﯽﻠﭘﻮﭙﯿﻟ ﺖﯿﻤﺳ ﯽﺳرﺮﺑ رﻮﻈﻨﻣ ﺲﮑﻠﭘ
ﺰﺘﻨﺳ يﺎﻫ هﺪﻧز ناﺰﯿﻣ و هﺪﺷ
لﻮﻠﺳ نﺪﻧﺎﻣ ﺶﯾﺎﻣزآ زا ﺎﻫ
فﺮﻌﻣ زا هدﺎﻔﺘﺳا ﺎﺑ MTT
3 - ) 4 و 5 - يد
(ﻞﯿﻟوزاﺮﺘﺗ ﻞﯿﺘﻣ -
2 و 5 مﻮﯿﻟوزاﺮﺘﺗ ﻞﯿﻨﻓ يد– ،ﺪﺷ هدﺎﻔﺘﺳا
ﮏﯾ MTT
ﮓﻧر شور لﺎﺘﺴﯾﺮﮐ ﻞﯿﮑﺸﺗ ﻪﯾﺎﭘ ﺮﺑ ﯽﺠﻨﺳ
لﻮﻠﺳ رد نازﺎﻣرﻮﻓ يﺎﻫ يﺎﻫ
ﺖﻗد و يﺮﯾﺬﭘراﺮﮑﺗ ﺖﯿﻠﺑﺎﻗ زا و ﺖﺳا هﺪﻧز يﺮﯿﮕﻤﺸﭼ
رادرﻮﺧﺮﺑ
.ﺖﺳا زا ﺮﻤﯿﻠﭘﻮﭙﯿﻟ ﺖﺒﺴﻧ ﺐﺴﺣ ﺮﺑ هﺪﺷ ﺰﺘﻨﺳ يﺎﻫ ﺮﻤﯿﻠﭘ ﯽﻧزو
ﻪﺑPEI
ﺪﯿﻤﺳﻼﭘ
(C/P)
ﺮﯾدﺎﻘﻣ ياﺮﺑ 4
/0
، 8 /0 و 2/
1 ) (w/w
ﻪﺑ رﻮﻈﻨﻣ
ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﯽﻠﭘﻮﭙﯿﻟ .ﺪﺷ هدﺎﻔﺘﺳا ﻒﻠﺘﺨﻣ يﺎﻫ ﺲﮑﻠﭘ
يوﺎﺣ ﺎﻫ
4µg
/0 ﺖﯿﻠﭘ ﮏﻫﺎﭼ ﺮﻫ يازا ﻪﺑ ﺪﯿﻤﺳﻼﭘ 96
رد ﻪﻧﺎﺧ يﺎﻫC/P
نﺎﯿﺑ
هﺪﺷ ﻮﭙﯿﻟ .ﺪﻧدﻮﺑ ﯽﻠﭘ ﺲﮑﻠﭘ ﯽﺘﻧآ نوﺪﺑ ﺖﺸﮐ ﻂﯿﺤﻣ رد ﺎﻫ رد ﮏﯿﺗﻮﯿﺑ
ﻢﺠﺣ 20µl
زا ﺲﭘ و ﻪﯿﻬﺗ 20
ﻪﺑ ﻂﯿﺤﻣ يﺎﻣد رد نﻮﯿﺳﺎﺑﻮﮑﻧا ﻪﻘﯿﻗد
زا ﺲﭘ .ﺪﯾدﺮﮔ ﻪﻓﺎﺿا ﮏﻫﺎﭼ ﺮﻫ رﺎﻬﭼ
ﺖﻋﺎﺳ ﻂﯿﺤﻣ ﺖﺸﮐ لﻮﻠﺳ ﺎﻫ
ﺧ راﺪﻘﻣ و ﺪﺷ جرﺎ 100µl
ﯽﺘﻧآ و مﺮﺳ يوﺎﺣ مﺮﮔ ﻂﯿﺤﻣ ﻪﺑ ﮏﯿﺗﻮﯿﺑ
تﺪﻣ ﻪﺑ ﺖﯿﻠﭘ .ﺪﯾدﺮﮔ ﻪﻓﺎﺿا ﮏﻫﺎﭼ ﺮﻫ 24
ﻮﮑﻧا نورد ﺖﻋﺎﺳ رﻮﺗﺎﺑ
ﻪﺑ .ﺪﺷ هداد راﺮﻗ لﻮﻠﺳ ﺖﺸﮐ ﮏﯿﻟﻮﺑﺎﺘﻣ ﺖﯿﻟﺎﻌﻓ ﯽﺳرﺮﺑ رﻮﻈﻨﻣ
(MTT)
راﺪﻘﻣ 20µl
فﺮﻌﻣ زا ﺖﻈﻠﻏ ﺎﺑMTT
mg/ml
5 ﻪﻓﺎﺿا ﮏﻫﺎﭼ ﺮﻫ ﻪﺑ
ﺪﺷ تﺪﻣ ﻪﺑ و ود
رد ﺖﻋﺎﺳ
°C
37 لﻮﻠﺤﻣ نآ زا ﺲﭘ .ﺪﯾدﺮﮔ ﻪﺑﻮﮑﻧا
ﻰﯾور ار ﻪﺑ رﻮﻃ ﻞﻣﺎﮐ راﺪﻘﻣ و هدﻮﻤﻧ جرﺎﺧ 100µl
ﻪﺑDMSO
رﻮﻈﻨﻣ
لﺎﺘﺴﯾﺮﮐ ﻞﺣ يﺎﻫ
ﻪﺑ ،هﺪﺷ ﻞﯿﮑﺸﺗ نازﺎﻣرﻮﻓ ،ﺪﺷ ﻪﻓﺎﺿا ﮏﻫﺎﭼ ﺮﻫ
زا ﺲﭘ ﺞﻨﭘ ﻪﻘﯿﻗد
، بﺬﺟ رد لﻮﻃ جﻮﻣ 540nm
زا هدﺎﻔﺘﺳا ﺎﺑ هﺎﮕﺘﺳد
ﺖﯿﻠﭘ رﺪﯾر
(Bio Tek epoch plate reader, USA)
ﯽﺳرﺮﺑ ﺪﯾدﺮﮔ .
ﻪﺑ ﯽﺒﺴﻧ ﻢﺴﯿﻟﻮﺑﺎﺘﻣ ﺖﯿﻟﺎﻌﻓ لﻮﻠﺳ بﺬﺟ ﺪﺻرد ترﻮﺻ
هﺪﺷ رﺎﻤﯿﺗ يﺎﻫ
لﻮﻠﺳ ﻪﺑ ﺖﺒﺴﻧ ﻞﻣﺎﺣﻮﻧﺎﻧ ﺎﺑ نﺎﯿﺑ لﺮﺘﻨﮐ يﺎﻫ
.ﺪﺷ
ﺶﯾﺎﻣزآ يﺎﻫ ﺎﺑ ﻪﺑﺎﺸﻣ ﻂﯾاﺮﺷ رد (ﯽﻧژ هدﺎﻣ لﺎﻘﺘﻧا) ﻦﺸﮑﻔﺴﻧاﺮﺗ
راﺪﻘﻣ نﺎﻤﻫ ﻮﭙﯿﻟ
ﯽﻠﭘ ﺲﮑﻠﭘ ﺖﯿﻤﺳ ﺶﯾﺎﻣزآ ياﺮﺑ هدﺎﻔﺘﺳا درﻮﻣ يﺎﻫ
تروﺎﺠﻣ زا ﺲﭘ ﯽﻟﻮﻠﺳ ﺖﯿﻤﺳ ﺶﯾﺎﻣزآ ﺪﻨﻧﺎﻤﻫ .ﺖﻓﺮﮔ مﺎﺠﻧا ﯽﻟﻮﻠﺳ 4 ﯽﻠﭘ ﻪﺘﻋﺎﺳ ﺲﮑﻠﭘ لﻮﻠﺳ ﺎﺑ ﺎﻫ
، نآ ﻂﯿﺤﻣ ﺾﯾﻮﻌﺗ ﻞﻣﺎﮐ ﻂﯿﺤﻣ ﺎﺑ ﺎﻫ
زا ﺲﭘ .ﺪﯾدﺮﮔ 24
ﺖﻋﺎﺳ لﻮﻠﺳ ﻂﯿﺤﻣ لﻮﻠﺳ و ﺪﺷ جرﺎﺧ ﺎﻫ
ﺎﺑ ﺎﻫ
زا هدﺎﻔﺘﺳا 100 µl
ﺮﻓﺎﺑ هﺪﻨﻨﮐﺰﯿﻟ
(Promega GmbH, Germany)
ﺗوﺮﭘ يﻮﺘﺤﻣ ﺎﺗ ﺪﻧﺪﺷ ﺐﯾﺮﺨﺗ ﯿ
نآ ﯽﻨﯿ رد ﺖﻨﺳرﻮﻠﻓ بﺬﺟ .دﻮﺷ دازآ ﺎﻫ
جﻮﻣ لﻮﻃ 460 nm
/ 510 ﺖﻨﺳرﻮﻠﻓ هﺎﮕﺘﺳد زا هدﺎﻔﺘﺳا ﺎﺑ
ﺮﺘﻣﻮﺘﻓوﺮﺘﮑﭙﺳا
(Spectrophotometer, PerkinElmer®, USA)
درﻮﻣ
.ﺖﻓﺮﮔ راﺮﻗ ﯽﺳرﺮﺑ لﻮﻠﺳ زا
ﺪﻧدﻮﺑ هﺪﺷ ﺖﮑﻔﺴﻧاﺮﺗ ﻪﮐ ﯽﮕﻧر ﺰﺒﺳ يﺎﻫ
ﺖﻨﺳرﻮﻠﻓ پﻮﮑﺳوﺮﮑﯿﻣ زا هدﺎﻔﺘﺳا ﺎﺑ ﺰﯿﻧ
(Nikon, Japan)
.ﺪﺷ يرادﺮﺒﺴﮑﻋ
هداد ﺮﺗ يﺎﻫ ا ﻪﺑ تارذ ﺰﯾﺎﺳ و ﯽﻟﻮﻠﺳ ﺖﯿﻤﺳ ،ﻦﺸﮑﻔﺴﻧ ترﻮﺻ
ﻦﯿﮕﻧﺎﯿﻣ ﻪﺑ راﺮﮑﺗ ﻪﺳ هاﺮﻤﻫ
±SD
ﻂﺳﻮﺗ يرﺎﻣآ ﻪﺴﯾﺎﻘﻣ و ﺪﻧا هﺪﺷ نﺎﯿﺑ
ﻪﻓﺮﻃ ﮏﯾ ﺲﻧﺎﯾراو
(one-way ANOVA)
.ﺪﺷ مﺎﺠﻧا ﺺﺨﺸﻣ ﺖﻬﺟ
هوﺮﮔ ﻦﯿﺑ فﻼﺘﺧا ندﻮﻤﻧ نﻮﻣزآ زا ﺎﻫ
Tukey Kramer
.ﺪﺷ هدﺎﻔﺘﺳا
ﻨﻌﻣ يرﺎﻣآ ﺮﻈﻧ زا يﺮﯾدﺎﻘﻣ ﺎ
ﻪﺟرد ،ﻞﻗاﺪﺣ ﻪﮐ ﺪﺷ ﻪﺘﻓﺮﮔ ﺮﻈﻧ رد راد
Downloaded from tumj.tums.ac.ir at 13:01 IRST on Friday December 18th 2020
ﺳرﺮﺑ ﯽ ﻟﯿ ﻠﭘﻮﭙ ﯿ ﺎﻫﺮﻤ اﺮﺑ ي تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ لﻮﻠﺳ ﻪﺑ نژ لﺎﻘﺘﻧا 287
نﺎﻨﯿﻤﻃا 95
% ﻪﺘﺷاد ار ﻪﺑ .ﺪﻨﺷﺎﺑ ﻦﯾا رد ،ﺮﮕﯾد ترﺎﺒﻋ ﺶﻫوﮋﭘ
، ﻦﯾﺮﺘﺸﯿﺑ
نﺎﻨﯿﻤﻃا مﺪﻋ ناﺰﯿﻣ
(Probability)
ﺮﯾﺬﭘ هﺪﺷ ﻪﺘﻓ 5
% ) 05 /0
P<
(
ﯽﻣ .ﺪﺷﺎﺑ ﻞﻣﺎﺣﻮﻧﺎﻧ ﯽﮔﮋﯾو ﻪﮐ ﯽﯾﺎﻫ ﻘﻣ هزاﺪﻧا ﺪﻨﻧﺎﻣ بﻮﻠﻄﻣ ﯽﮑﯾﺰﯿﻓ يﺎﻫ
سﺎﯿ
ﻮﻧﺎﻧ و ﺎﺗﯾ ﯿﺪ لﺎﺼﺗا و هﺪﺷ ﻪﻌﻟﺎﻄﻣ دراو ﺪﻨﺷﺎﺑ ﻪﺘﺷاد ار PEI
ﻗ ﺖﯿﻠﺑﺎ
ﻞﻣﺎﺣﻮﻧﺎﻧ ﺖﯿﻤﺳ و ﻦﺸﮑﻔﺴﻧاﺮﺗ راﺮﻗ ﯽﺳرﺮﺑ درﻮﻣ ﺎﻫ
.ﺖﻓﺮﮔ
ﻮﻧﺎﻧ ﺎﻫرﺎﺘﺧﺎﺳ زا ﺮﺘﮔرﺰﺑ ي 200 nm
ﻣ زا هدﻮﻟآ ﯽﻟﻮﻠﺳ ﺖﯿﻠﭘ و ﻟﺎﻄ
ﻪﻌ
.ﺪﺷ فﺬﺣ
ﻦﯿﻤﯾا ﻦﻠﯿﺗاﯽﻠﭘ ﺮﻤﯿﻠﭘ ﺐﯿﮐﺮﺗ ﺎﺑ ﯽﯾﺎﻫﺲﮑﻠﭘﯽﻠﭘﻮﭙﯿﻟ ،ﻪﻌﻟﺎﻄﻣ ﻦﯾا رد تارذ هزاﺪﻧا .ﺪﯾدﺮﮔ ﻞﺻﺎﺣ ﺪﯿﻤﺳﻼﭘ ﺎﺑ موزﻮﭙﯿﻟ و 1nm
/ 104 ﺑ ﻪ ﺖﺳد
ﯽﻠﭘﻮﭙﯿﻟ هزاﺪﻧا .ﺪﻣآ ﻪﺑ موزﻮﭙﯿﻟ و ﺮﻤﯿﻠﭘ ﺎﺑ ﻪﺴﯾﺎﻘﻣ رد ﺲﮑﻠﭘ
ﺗ ﺎﻬﻨ ﯽﯾ
98nm
نﺎﺸﻧ ار ﺶﯾاﺰﻓا ﯽﻣ
زا ﺮﺘﻤﮐ نآ ﺮﻄﻗ ﺎﻣا ﺪﻫد 200nm
ﯽﻣ .ﺪﺷﺎﺑ
ﻪﺑ ﻪﻨﯿﻬﺑ رﻮﻈﻨﻣ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺪﻨﯾاﺮﻓ يزﺎﺳ
يداﺪﻌﺗ ﺎﻣﺪﻘﻣ تﺎﺸﯾﺎﻣزآ ﺑ ﯽﺗ
ﺮ
لﻮﻠﺳ يور يﺎﻫ
HEK-293
ﻪﺑ ﻪﮐ ﺖﻟﻮﻬﺳ ﻪﺑ ﻪﮐ ﯽﻟﻮﻠﺳ هدر ناﻮﻨﻋ
ﻞﻣﺎﺣ ﯽﻣ ار ﯽﻧژ يﺎﻫ ﺮﯾﺬﭘ
ﻧ ﻪﺑ و ﺪ ﯽﺳرﺮﺑ ياﺮﺑ يدراﺪﻧﺎﺘﺳا ناﻮﻨﻋ
ﯽﻣ هدﺎﻔﺘﺳا ﻦﺸﮑﻔﺴﻧاﺮﺗ ﯽﯾارﺎﮐ ﻮﺷ
ﻧ ﺻ ،ﺪ ﺖﻓﺮﮔ ترﻮ .
12
ﺗوﺮﭘ نﺎﯿﺑ ﯿ ﮓﻧر ﺰﺒﺳ ﻦﯿ لﻮﻠﺳ ردGFP
ﺎﺑ هﺪﺷ ﺖﮑﻔﺴﻧاﺮﺗ يﺎﻫ
ﺪﺷ ﯽﺳرﺮﺑ ﺮﺘﻤﯾرﻮﻠﻓ هﺎﮕﺘﺳد ﺶﯾﺎﻣزآ ﻪﻠﻤﺟ زا .
ﻪﻨﯿﻬﺑ يﺎﻫ يزﺎﺳ
ﯽﻣ نژ نﺎﯿﺑ ﯽﺳرﺮﺑ ياﺮﺑ ﺐﺳﺎﻨﻣ نﺎﻣز ﯽﺳرﺮﺑ ﻪﺑ ناﻮﺗ ﯾ رﻮﻀﺣ ،GFP
ﺎ
رد مﺮﺳ رﻮﻀﺣ مﺪﻋ لﻮﻃ
رﺎﻬﭼ ﯽﻠﭘﻮﭙﯿﻟ تروﺎﺠﻣ ﺖﻋﺎﺳ ﺎﺑ ﺲﮑﻠﭘ
ﻮﭙﯿﻟ ﻪﯿﻬﺗ هﻮﺤﻧ و لﻮﻠﺳ ﯽﻠﭘ
ﻪﮐ ﯽﻧﺎﻣز ﻦﯾﺮﺘﻬﺑ .دﻮﻤﻧ هرﺎﺷا ﺲﮑﻠﭘ
لﻮﻠﺳ يﺎﻫ ﯽﻣ نﺎﺸﻧ ار هﺪﺷ ﻞﻘﺘﻨﻣ نژ نﺎﯿﺑHEK
ﺪﻧداد 48 ﻮﺑ ﺖﻋﺎﺳ .د
ﻪﭼﺮﮔ ﻪﺑ ﻣ ﺮﻈﻧ ﯽ ﺳر ﯿ ﺪ لﻮﻠﺳ تروﺎﺠﻣ رد مﺮﺳ رﻮﻀﺣ رد ﻪﮐ ﯽﻧﺎﻣز ﺎﻫ
ﯽﻠﭘﻮﭙﯿﻟ ﺪﻨﺘﺷاد راﺮﻗ ﺲﮑﻠﭘ
، ﯽﻣ نﺎﺸﻧ يﺮﺘﻬﺑ ﺎﻘﺑ ﻨﻌﻣ) ﺪﻨﻫد
ﺎ ﻧ راد ﻮﺒ ﺎﻣا (د
ﻪﺑ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺖﯿﻠﺑﺎﻗ يﺮﯿﮕﻤﺸﭼ زﺮﻃ
ﺖﻓﺎﯾ ﺶﻫﺎﮐ .
ﻦﯾازا ور رد
ﺶﻫوﮋﭘ يﺎﻫ ﯽﺗآ لﻮﻠﺳ يور ﺮﺑ يﺎﻫ
ﯽﻠﭘﻮﭙﯿﻟ ،PC3
ﺲﮑﻠﭘ مﺪﻋ رد ﺎﻫ
مﺮﺳ رﻮﻀﺣ
، تﺪﻣ ﻪﺑ رﺎﻬﭼ .ﺪﻧﺪﺷ روﺎﺠﻣ فﺪﻫ لﻮﻠﺳ ﺎﺑ ﺖﻋﺎﺳ
ﯽﻠﭘﻮﭙﯿﻟ ﻪﯿﻬﺗ درﻮﻣ رد درﻮﻣ ﺖﻟﺎﺣ ود ﺲﮑﻠﭘ
.ﺖﻓﺮﮔ راﺮﻗ ﯽﺳرﺮﺑ
ﻪﮑﻨﯾا لوا ﺖﻟﺎﺣ :
"
ﻠﭘﯿ ﺮﻤ ﻤﺳﻼﭘ ﺎﺑ ﯿ ﺪ ﻠﭘ ، ﯽ ﺲﮑﻠﭘ ﮑﺸﺗ ﯿ ﻞ ﺲﭙﺳ و ﺪﻫﺪﺑ
ﮐﺮﺗ رد ﯿ ﺐ ﻟ ﺎﺑ ﯿ موزﻮﭙ ﻟ ، ﯿ ﻠﭘﻮﭙ ﯽ ا ﺲﮑﻠﭘ ﯾ دﺎﺠ ﺪﻨﮐ ﻦﯾا ﺎﯾ و ﺖﻟﺎﺣ رد ﻪﮐ
ﺲﭙﺳ و هﺪﺷ ﺐﯿﮐﺮﺗ ﺮﮕﯾﺪﮑﯾ ﺎﺑ موزﻮﭙﯿﻟ و ﺮﻤﯿﻠﭘ اﺪﺘﺑا مود تروﺎﺠﻣ رد
ﻤﺳﻼﭘ ﯿ ﺪ ، ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﮑﺸﺗ ﯿ ﻞ ﺪﻨﻫد ) رادﻮﻤﻧ 1 .(
هداد يﺎﻫ
ﻨﻌﻣ توﺎﻔﺗ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺎ
ﻪﺟﻮﺗ ﺎﺑ و ﺪﻧاﺪﻧ نﺎﺸﻧ شور ود ﻦﯾا ﻦﯿﺑ يراد
ﺠﺗ و ﻪﺑﺎﺸﻣ تﻻﺎﻘﻣ ﻪﺑ ﺮﺑ تﺎﺸﯾﺎﻣزآ ﻪﻣادا ياﺮﺑ مود شور ،ﻪﺘﺷﺬﮔ ﻪﺑﺮ
لﻮﻠﺳ يور يﺎﻫ
ﻪﻨﯿﻬﺑ تﺎﺸﯾﺎﻣزآ زا ﺲﭘ ،ﺪﯾدﺮﮔ بﺎﺨﺘﻧا PC3
،يزﺎﺳ
ﺎﺑ ﯽﯾﺎﻫ C/P
ﺮﯾدﺎﻘﻣ 4 /0
، 8/
0 و 2/
1 ) (w/w
ﻦﺸﮑﻔﺴﻧاﺮﺗ ياﺮﺑ ﺰﯿﻧ
لﻮﻠﺳ يﺎﻫ
3
.ﺪﺷ ﻪﺘﻓﺮﮔ ﺮﻈﻧ ردPC
8
ﯽﻠﭘﻮﭙﯿﻟ ،داد نﺎﺸﻧ ﺞﯾﺎﺘﻧ ﻪﯾﺎﭘ ﺎﺑ ﺲﮑﻠﭘ
نزو ﺎﺑPEI
25 ﻮﻠﯿﮐ ،نﻮﺘﻟاد
رد ﺖﺒﺴﻧ 4 /0
= نوﺰﻓا CP
ﻦﯾا ﺮﺑ ﯾﺎﭘ ﻪﺑ ﺖﺒﺴﻧ يﺮﺘﻬﺑ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﻪﮐ ﻪ
) ﯽﻣوزﻮﭙﯿﻟﻮﻧﺎﻧ 05
/0
P<
ﯽﻣ نﺎﺸﻧ ( ﻪﮐ ﺖﻨﺳرﻮﻠﻓ ناﺰﯿﻣ ﻪﮑﻠﺑ ﺪﻫد
يﺮﻤﯿﻠﭘ ﻪﯾﺎﭘ ﻪﺑ ﺖﺒﺴﻧ ،ﺖﺳا نژ لﺎﻘﺘﻧا زا ﯽﺑﺎﺗزﺎﺑ ﻦﯿﻤﻫ ردPEI
CP
ﺶﯾاﺰﻓا 5/
2 ﺮﮕﯾد رد .ﺖﺳا ﻪﺘﺷاد يﺮﺑاﺮﺑ ﺎﻣ ﯽﺗوﺎﻔﺗ ﺎﻫ CP
ﯽﻠﭘﻮﭙﯿﻟ ﻦﯿﺑ ﺲﮑﻠﭘ
ﯽﻠﭘﻮﭙﯿﻟ و ﺪﺸﻧ هﺪﻫﺎﺸﻣ موزﻮﭙﯿﻟﻮﻧﺎﻧ و ﺖﺒﺴﻧ ﺲﮑﻠﭘ
ﻘﺗ .داد نﺎﺸﻧ ﻦﺸﮑﻔﺴﻧاﺮﺗ ياﺮﺑ يﺮﺘﻤﮐ ﯽﯾﺎﻧاﻮﺗ يﺮﻤﯿﻠﭘ ﻪﯾﺎﭘ ﻪﺑ ﺒﯾﺮ
ﺎ
مﺎﻤﺗ رد ﻪﺑ ﺎﻫCP
ﻦﯿﯾﺎﭘ ﺰﺟ ﻦﯾﺮﺗ ﯽﻟﻮﻠﺳ ﺎﻘﺑ ﺖﯿﻠﺑﺎﻗ ﺶﻫﺎﮐ CP
لﻮﻠﺳ رد موزﻮﭙﯿﻟﻮﻧﺎﻧ ﺎﺑ هﺪﺷ ﺖﮑﻔﺴﻧاﺮﺗ يﺎﻫ ﺮﻤﯿﻠﭘﻮﻧﺎﻧ ﻪﺑ ﺖﺒﺴﻧ
ي و
ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﻣ هﺪﻫﺎﺸ ) ﺪﺷ رادﻮﻤﻧ 2B
.(
رﺮﺑ و ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺞﯾﺎﺘﻧ ﯽﺳ
رد ﯽﻟﻮﻠﺳ ﺖﯿﻤﺳ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ ﻪﯾﺎﭘ ﺎﺑ نزو ﺎﺑPEI
10 ﺘﻟاد ﻮﻠﯿﮐ رد ،نﻮ
ﺖﺒﺴﻧ 4 /0
= ﻢﻫ CP
موزﻮﭙﯿﻟﻮﻧﺎﻧ ﻪﯾﺎﭘ ﻪﺑ ﺖﺒﺴﻧ يﺮﺘﻬﺑ ﻦﺸﮑﻔﺴﻧاﺮﺗ و
ﯽﻣ نﺎﺸﻧ ﺮﻤﯿﻠﭘﻮﻧﺎﻧ ﺪﻫد
) 001 /0
P<
( ﻢﻫ و ﻪﺑ نژ لﺎﻘﺘﻧا ناﺰﯿﻣ
ﯽﻠﭘ ﻪﺑ ﺖﺒﺴﻧ ،لﻮﻠﺳ ﺲﮑﻠﭘ
يﺎﻫ
25 kDa
ردPEI
8 /0
= ﻪﺑ ﻪﮐCP
ناﻮﻨﻋ
رادﻮﻤﻧ 1 : ﺳرﺮﺑ ﯽ ارﺎﮐ ﯽﯾ لﻮﻠﺳ رد ﻦﺸﮑﻔﺴﻧاﺮﺗ
HEK-293
ﻪﺘﻓﺎﯾ ﺎﻫ
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288
ﻒﺠﻧ يﺮﻔﻌﺟ ﻦﺴﺣ ﺪﻤﺤﻣ يدﺎﺑآ
نارﺎﮑﻤﻫ و
رادﻮﻤﻧ 2 : ﺳرﺮﺑ ﯽ ﻤﺳ ﯿ ﺖ ﻟﻮﻠﺳ ﯽ و(A)
ارﺎﮐ ﯽﯾ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﻟﻮﻠﺳ هدر رد(B)
ﯽ PC3
رادﻮﻤﻧ 3 : ﮐ ﯽﺳرﺮﺑ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﯽﯾارﺎ ﯽﻟﻮﻠﺳ ﺖﯿﻤﺳ و(A)
ﯽﻟﻮﻠﺳ هدر رد(B)
PC3
ﺶﯾﺎﻣزآ ياﺮﺑ دراﺪﻧﺎﺘﺳا يﺎﻫ
ﯽﻣ ﻪﺘﻓﺮﮔ ﺮﻈﻧ رد ﻦﺸﮑﻔﺴﻧاﺮﺗ ،دﻮﺷ
ﺶﯾاﺰﻓا 2 /1 ) ﺖﺳا ﻪﺘﺷاد يﺮﺑاﺮﺑ رادﻮﻤﻧ
3A
ﺮﮕﯾد رد .(
ﯽﺗوﺎﻔﺗ ﺎﻫCP
ﻦﯿﺑ ﺎﻣ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﺸﻧ هﺪﻫﺎﺸﻣ موزﻮﭙﯿﻟﻮﻧﺎﻧ و ﻦﯾاﺎﺑ .ﺪ
ﺖﯿﻤﺳ لﺎﺣ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ ﯽﺘﺣ و ﺖﺳا ﻪﺘﻓﺎﯾ ﺶﻫﺎﮐ ﯽﻣوزﻮﭙﯿﻟﻮﻧﺎﻧ ﻪﯾﺎﭘ ﻪﺑ ﺖﺒﺴﻧ ﺎﻫ
رد 4/
0 داد نﺎﺸﻧ ار يﺮﺘﻬﺑ ﯽﻟﻮﻠﺳ ﺎﻘﺑ ﻢﻫ يﺮﻤﯿﻠﭘ ﻪﯾﺎﭘ ﻪﺑ ﺖﺒﺴﻧC/P=
) رادﻮﻤﻧ 3B
.(
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ﺳرﺮﺑ ﯽ ﻟﯿ ﻠﭘﻮﭙ ﯿ ﺎﻫﺮﻤ اﺮﺑ ي تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ لﻮﻠﺳ ﻪﺑ نژ لﺎﻘﺘﻧا 289
ﻞﮑﺷ 1 : لﻮﻠﺳ زا يﺮﯾﻮﺼﺗ ﯽﻠﭘﻮﭙﯿﻟ ﺎﺑ هﺪﺷ ﺖﮑﻔﺴﻧاﺮﺗ يﺎﻫ
ﺲﮑﻠﭘ ﻪﯾﺎﭘ يﺎﻫ 10PEI
و 25 ﻮﻠﯿﮐ اد نﻮﺘﻟ
ﻪﻌﻟﺎﻄﻣ رد يا
،ﺪﻧداد نﺎﺸﻧ ﺶﻧارﺎﮑﻤﻫ و EWE
رد
ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ زا هﺪﺷ ﻞﯿﮑﺸﺗ يﺎﻫ ﻞﮑﺷ تارذﻮﻧﺎﻧ ،موزﻮﭙﯿﻟ و PEI
ﺖﺧاﻮﻨﮑﯾ ﻪﯾﺰﺠﺗ ﻪﺑ ،نﺎﺷ هﺪﻧزﺎﺳ ياﺰﺟا ﺎﺑ ﻪﺴﯾﺎﻘﻣ رد و ﺪﻨﺘﺷاد يﺮﺗ
(Aggregation)
ﺮﮕﯾد فﺮﻃ زا .ﺪﻧداد نﺎﺸﻧ يﺮﺘﻬﺑ ﺖﻣوﺎﻘﻣ
ﻠﭘﻮﭙﯿﻟ ﯽ ﺲﮑﻠﭘ يزرﻮﺘﺳد ﻪﺑ ﺎﻫ ﯽﻠﭘ ﻪﺑ لﺎﺼﺗا ﻞﺜﻣ يﺮﺘﻤﮐ يﺎﻫ
ﻦﻠﯿﺗا
لﻮﮑﯿﻠﮔ
(PEG)
دﻮﺒﻬﺑ ياﺮﺑ ﯽﮔﮋﯾو
يﺎﻫ ﮑﯾﺰﯿﻓ ﯽ ، ﮏﯾژﻮﻟﻮﯿﺑ و ﯽﯾﺎﯿﻤﯿﺷ
ﺪﻨﺘﺷاد زﺎﯿﻧ .
ﻦﯾا 10
زا هوﺮﮔ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ يﺎﻫ ﻪﺑ ﺖﺳد هﺪﻣآ ﻪﺑ رﻮﻈﻨﻣ
لﺎﻘﺘﻧا
siRNA
اﺮﺑ ي نژ ندﺮﮐ شﻮﻣﺎﺧ ﺎﻫ
ي هدﺎﻔﺘﺳا رﻮﻣﻮﺗ ﺪﻧﺪﺷ
ﻟﺎﺣرد ﯽ ﻪﮐ رد ا ﻦﯾ ﻟﺎﻄﻣ ﻟ زا ﻪﻌ ﺲﮑﻠﭘﻮﭙﯿ ﻪﺑ لﺎﻘﺘﻧا رﻮﻈﻨﻣ ﺪﯿﻤﺳﻼﭘ
يوﺎﺣ
نژ ﺮﮔﺎﺳﺎﻨﺷ ﻧﺎﻃﺮﺳ لﻮﻠﺳ ﻪﺑ ﯽ
هدﺎﻔﺘﺳا توﺎﻔﺗ دﻮﺟو ﺎﺑ ﻪﺘﺒﻟا ،ﺪﺷ ﺎﻫ
ﯽﯾ
ﻬﺑ رد ﻪﻨﯿ زﺎﺳ ي ﺎﻣزآ ﺶﯾ ﻪﺑ ﻣ ﺮﻈﻧ ﯽ ﺮﻫ ﺪﺳر ﺳا ود
ﺪﯿ ﺌﻠﮐﻮﻧ ﮏﯿ زا ﻧﺎﮑﻣ ﻢﺴﯿ اﺮﺑ ﻪﺑﺎﺸﻣ ي ﻪﺑ دورو لﻮﻠﺳ
هدﺎﻔﺘﺳا ﯽﻣ ﺪﻨﻨﮐ ﻪﭼﺮﮔ .
Haghiralsadat
و ﺶﻧارﺎﮑﻤﻫ نﺎﺸﻧ
لﺎﺼﺗا ﺪﻧداد ﻞﻣﺎﺣ ﻪﺑ PEG
يﺎﻫ
ﯽﻣوزﻮﭙﯿﻟ ﻪﺑ ﺖﯿﻟﺎﻌﻓ و نﻮﺧ شدﺮﮔ نﺎﻣز ،ﺐﯿﮐﺮﺗ يراﺪﯾﺎﭘ دﻮﺒﻬﺑ ﺐﺒﺳ
ﯽﻣ دﺎﻬﻨﺸﯿﭘ ﯽﺘﺴﯾز ﻦﯾا ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﺎﻣا دﻮﺷ
دﻮﺧ ﻪﮐ ﺐﺒﺳ PEG
ﺦﺳﺎﭘ ﯽﻣ ﯽﻨﻤﯾا يﺎﻫ ددﺮﮔ
ا رد ﻦﯾ ﺎﻄﻣ ﻪﻌﻟ زا ﻞﻣﺎﺣﻮﻧﺎﻧ رﺎﺘﺧﺎﺳ ردPEG
دﺮﮕﻧ هدﺎﻔﺘﺳا ﺪﯾ
.
13 و
ﻪﻌﻟﺎﻄﻣ رد 14
يا ﻧ ﺶﻧارﺎﮑﻤﻫ و Lee
،ﺪﻧداد نﺎﺸ
ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﯽﻣ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﯽﯾارﺎﮐ ﺶﯾاﺮﻓا ﺐﺒﺳ ﺎﻫ ﺪﻧﻮﺷ
و يراﺪﯾﺎﭘ
نآ ﯽﻧﻻﻮﻃ هﺮﯿﺧذ رد ﺎﻫ ﺖﺳا ﺮﺘﺸﯿﺑ تﺪﻣ
. رد ا ﻪﻌﻟﺎﻄﻣ نﺎﺸﯾ زا
PEI
ﺎﻫ ﯽﯾ ﻟﻮﮑﻟﻮﻣ نزو ﺎﺑ ﯽ
8 /0 و 25 ﮐ ﻮﻠﯿ نﻮﺘﻟاد و
Poly-L-lysine
نزو ﺎﺑ
ﻟﻮﮑﻟﻮﻣ ﯽ 30 - 15 ﮐ ﺐﯿﮐﺮﺗ رد نﻮﺘﻟادﻮﻠﯿ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ هدﺎﻔﺘﺳا درﻮﻣ
ﺖﻓﺮﮔ راﺮﻗ .
ﺖﻬﺑﺎﺸﻣ ا ﻦﯾ ﺮﺿﺎﺣ ﻪﻌﻟﺎﻄﻣ ﺎﺑ ﻪﻌﻟﺎﻄﻣ رد
زا هدﺎﻔﺘﺳا ﺎﺑPEI
ﻟﻮﮑﻟﻮﻣ نزو ﯽ
25 ﮐ نﻮﺘﻟادﻮﻠﯿ ﺖﺳا . ﮐﺮﺗ ﺖﺒﺴﻧ ﻪﺘﺒﻟا ﯽﺒﯿ
ﻟ ﺎﺑ نآ موزﻮﭙﯿ و
ﻣ توﺎﻔﺘﻣ فﺪﻫ لﻮﻠﺳ عﻮﻧ ﯽ
ﺪﺷﺎﺑ ﻨﭽﻤﻫ و ﯿ ﻦ ا رد ﯾ ﻦ زا ﻪﻌﻟﺎﻄﻣ ﺎﺑPEI
ﺚﺤﺑ
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290
ﻒﺠﻧ يﺮﻔﻌﺟ ﻦﺴﺣ ﺪﻤﺤﻣ يدﺎﺑآ
نارﺎﮑﻤﻫ و
ﻟﻮﮑﻟﻮﻣ نزو ﯽ
ﻂﺳﻮﺘﻣ 10 ﮐ ﯿ نﻮﺘﻟادﻮﻠ ﻤﺳ ﻪﮐ ﯿ ﺖ ﺮﺘﻤﮐ ي ﻪﺑ ﺖﺒﺴﻧ 25
ﮐﯿ ﺘﻟادﻮﻠ نﻮ ﻧ دراد ﯿﺰ ﺖﺳا هﺪﺷ هدﺎﻔﺘﺳا .
ﻪﮐ ﺖﺷاد ﻪﺟﻮﺗ ﺪﯾﺎﺑ ﺎﻣا 15
ﻢﮐاﺮﺘﻣ رد ﺮﻤﯿﻠﭘ ﻒﯿﻌﺿ ﯽﺘﺴﯾز ﺖﯿﻟﺎﻌﻓ ﯽﻤﻧ ﺪﯿﺳا ﮏﯿﺌﻠﮐﻮﻧ يزﺎﺳ
ﺪﻧاﻮﺗ
ﻂﺳﻮﺗ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ دﻮﺷ ناﺮﺒﺟ .
نﺎﺸﻧ ﻪﺘﺷﺬﮔ تﺎﻌﻟﺎﻄﻣ ﻪﮐ رﻮﻄﻧﺎﻤﻫ16
ﺎﺑ لﺎﻘﺘﻧا رد نژ يﻻﺎﺑ نﺎﯿﺑ ،ﺪﻧداد ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ ﻪﺑ ﻻﺎﻤﺘﺣا ﺎﻫ ﺮﺛا ﻞﯿﻟد
ﺧ هﺪﻨﯾاﺰﻓا ﯽﻣوزوﺪﻧآ زا راﺮﻓ ﺖﯿﺻﻮﺼﺧ و ﯽﻧﻮﺗوﺮﭘ ﺞﻨﻔﺳا ﺖﯿﺻﺎ
ﯽﻠﭘ و ﯽﻧﻮﯿﺗﺎﮐ ﺪﯿﭙﯿﻟ ﻂﺳﻮﺗ ﻪﮐ ﺖﺳا ﯽﻣ ﻢﻫاﺮﻓ ﻦﯿﻤﯾا ﻦﻠﯿﺗا
دﻮﺷ .
17
ﻦﯾاﺎﺑ ﻪﭼﺮﮔ ،لﺎﺣ ) ﻻﺎﺑ ﯽﻟﻮﮑﻟﻮﻣ نزو ﺎﺑPEI
25 ﻪﺑ (نﻮﺘﻟادﻮﻠﯿﮐ ﯽﺑﻮﺧ
ﯽﻣ لﺎﻌﻓ ار ﯽﻧﻮﺗوﺮﭘ ﺞﻨﻔﺳا هﺪﯾﺪﭘ ﺖﺴﯾز ﺎﻣا ﺪﻨﮐ
و ﺖﺴﯿﻧ رﺎﮔزﺎﺳ
ﯽﻟﻮﻠﺳ ﺖﯿﻤﺳ ﻪﺑ
ار ﯽﯾﻻﺎﺑ ﺖﺒﺴﻧ ﯽﻣ نﺎﺸﻧ
ﺪﻫد . ﻪﺑ18
ﯽﻣ ﺮﻈﻧ ﺖﯿﻤﺳ ﺪﺳر
ﯽﻟﻮﮑﻟﻮﻣ نزو ﻪﺑ ﯽﻟﻮﻠﺳ دراد ﯽﮕﺘﺴﺑ فﺪﻫ لﻮﻠﺳ عﻮﻧ و PEI
.
19
ﻞﻣﺎﺣﻮﻧﺎﻧ ﺖﯿﻤﺳ شور ﺎﺑ هﺪﺷ ﺰﺘﻨﺳ يﺎﻫ
ﺖﺒﺴﻧ رد و MTT
يﺎﻫ
ﺪﯿﻤﺳﻼﭘ ﻪﺑ ﻞﻣﺎﺣ زا ﻒﻠﺘﺨﻣ
(C/P)
ﺎﺑ ﺮﯾدﺎﻘﻣ 4 /0
، 8/
0 و 2/
1 ) (w/w
هدر رد تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ لﻮﻠﺳ ﺮﮔ راﺮﻗ ﯽﺑﺎﯾزرا درﻮﻣ PC3
ﺖﻓ
) رادﻮﻤﻧ 2 وA
3 (.B
ﻟﯿ موزﻮﭙ ﺎﻫ ﻤﺳ ﯿ ﺖ ﻟﻮﻠﺳ ﯽ ﻻﺎﺑ ﯽﯾ ﻣ نﺎﺸﻧ ﯽ ﺪﻨﻫد ﻪﮐ
ﻻﺎﺑ رﺎﺑ ﺎﺑ ﻂﺒﺗﺮﻣ ﻻﺎﻤﺘﺣا ي
ﻣ ﺎﻬﻧآ ﯽ ﺪﺷﺎﺑ ﺮﻣا ﻦﯾا ﻪﮐ ﺐﺒﺳ
ﺶﻨﮑﻤﻫﺮﺑ ﻟﻮﻠﺳ يﺎﺸﻏ ﺎﺑ يﻮﻗ يﺎﻫ ﯽ
ﯽﻣ لﻮﻠﺳ ﻪﺑ ﺐﯿﺳآ و دﻮﺷ
.
20
ﯽﻣﺎﻤﺗ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﺑ ﺎﻫ ﺮ ودﺮﻫ ﻪﯾﺎﭘ لﻮﻠﺳ ﺎﻘﺑ رد يدﻮﺒﻬﺑ PEI
ار
ﺸﻧ ﻪﯾﺎﭘ موزﻮﭙﯿﻟ ﻪﺑ ﺖﺒﺴﻧ ﯽﻣ نﺎ
.ﺪﻨﻫد ﻟﯿ ﻠﭘﻮﭙ ﯽ ﺲﮑﻠﭘ ﺎﻫ ي ﺎﭘ ﺎﺑ ﯾﻪ
PEI 25
ﺎﻘﻣ رد ﯾ ﻪﺴ ﺎﺑ
PEI 10
ﺑﯿ ﺮﺘﺸ ﻟﻮﻠﺳ ﺎﻘﺑ ﺶﻫﺎﮐ ﺚﻋﺎﺑ ﯽ
ﻣ ﯽ ) ﺪﻧﻮﺷ رادﻮﻤﻧ
3 (B
ا ﻪﮐ ﯾ ﻦ ﺮﻣا ﻪﺑ ﻟد ﯿ ﻞ ﻤﺳ ﯿ ﺖ ﺗاذ ﯽ ﻟﻮﮑﻟﻮﻣ نزو ﺎﺑPEI
ﯽ ﻪﺑ.ﺖﺳا ﻻﺎﺑ
د نﺎﺑز ﯾ ﺮﮕ ﻤﺳ ﯿ ﺖ ﻟﻮﻠﺳ ﯽ ﻻﺎﺑ ي ﻻﺎﺑ رﺎﺑ ﺎﺑ PEI
ي اﯾ ﻦ ﻠﭘﯿ ﺮﻤ نزو و
ﻟﻮﮑﻟﻮﻣ ﯽ ﻘﺘﺴﻣ طﺎﺒﺗرا نآ ﯿ
ﻢ د درا . لﻮﻠﺳ رد ﻦﺸﮑﻔﺴﻧاﺮﺗ21
ﻫ نﺎﻃﺮﺳ يﺎ
تﺎﺘﺳوﺮﭘ و ﺪﺷ مﺎﺠﻧا ﯽﻟﻮﻠﺳ ﺖﯿﻤﺳ ﺶﯾﺎﻣزآ ﺎﺑ ﻪﺑﺎﺸﻣ ﻂﯾاﺮﺷ ردPC3
ﺗوﺮﭘ نﺎﯿﺑ ﯿ ﮓﻧر ﺰﺒﺳ ﻦﯿ لﻮﻠﺳ ردGFP
هﺎﮕﺘﺳد ﺎﺑ هﺪﺷ ﺖﮑﻔﺴﻧاﺮﺗ يﺎﻫ
.ﺪﺷ ﯽﺳرﺮﺑ ﺮﺘﻤﯾرﻮﻠﻓ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ ﺮﻫ ﺎﺑ هﺪﺷ ﺰﺘﻨﺳ يﺎﻫ ﻪﯾﺎﭘ ود
PEI
رد 4 /0 ﻪﺑ ﺖﺒﺴﻧ ار ﻦﺸﮑﻔﺴﻧاﺮﺗ ﻦﯾﺮﺘﺸﯿﺑC/P=
موزﻮﭙﯿﻟ و PEI
ﻪﯾﺎﭘ
هداد نﺎﺸﻧ ا
ﺪﻧ ﻞﮑﺷ) 1 ( ﺖﯿﻤﺳ ﺶﻫﺎﮐ ﺎﺑ ﻻﺎﻤﺘﺣا ﺮﻣا ﻦﯾا ،
ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﺎﻫ ﻪﺑ ﺖﺒﺴﻧ ﻞﻣﺎﺣ ﻪﯾﺎﭘ يﺎﻫ ﺖﺳد هﺪﺸﻧ يزرو ﻦﯾا رد
ﺖﻈﻠﻏ
، ﯽﻣ ﻂﺒﺗﺮﻣ .ﺪﺷﺎﺑ
ﯽﻠﭘﻮﭙﯿﻟ رد ﺲﮑﻠﭘ
ﻪﯾﺎﭘ ﺎﺑ يﺎﻫ 10 PEI
نوﺰﻓا نﻮﺘﻟادﻮﻠﯿﮐ ﻦﯾا ﺮﺑ
ﯽﻠﭘ ﻪﺑ ﺖﺒﺴﻧ ار ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺶﯾاﺰﻓا ﻪﮐ ﺲﮑﻠﭘ
10PEI
ﺶﯾاﺰﻓا داد نﺎﺸﻧ (موزﻮﭙﯿﻟ نوﺪﺑ ﺺﻟﺎﺧ) 2/
1 ﺖﺒﺴﻧ يﺮﺑاﺮﺑ
ﯽﻠﭘ ﻪﺑ ﺲﮑﻠﭘ 25 ﻪﺑ ﻪﮐ) PEI
ﻪﺘﻓﺮﮔ ﺮﻈﻧ رد دراﺪﻧﺎﺘﺳا ﺪﻠﮔ ناﻮﻨﻋ
ﯽﻣ ﺪﺷ هﺪﻫﺎﺸﻣ ﻢﻫ ،(دﻮﺷ .
ﻪﯿﺿﺮﻓ ،ﺞﯾﺎﺘﻧ ﻦﯾا ﻪﻌﻟﺎﻄﻣ
ﯽﻧﻮﻨﮐ ﺎﺑ ﻪﮐ ار
ﯽﻣ ﺶﯾاﺰﻓا ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺖﯿﻠﺑﺎﻗ ،ﺖﯿﻤﺳ ﺶﻫﺎﮐ ﯽﻣ ﺪﯿﯾﺎﺗ ﺪﺑﺎﯾ
.ﺪﻨﮐ زا
ﯽﻣ ﻪﺑﺎﺸﻣ تﺎﻌﻟﺎﻄﻣ ناﻮﺗ
ﻪﺑ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﻪﯾﺎﭘ ﺎﺑ ﯽﯾﺎﻫ 10 PEI
يوﺎﺣ موزﻮﭙﯿﻟ و نﻮﺘﻟادﻮﻠﯿﮐ
DOTAP
ﯽﻠﭘ ،لوﺮﺘﺴﻠﮐ ، لﻮﮑﯿﻠﮔ ﻦﻠﯿﺗا
(PEG)
و ﻪﺑ DSPE
لﺎﻘﺘﻧا رﻮﻈﻨﻣ ﺮﯿﻏ RNA
ﯽﺳﺪﻨﻬﻣ هﺪﻨﻨﮐﺪﮐ هﺪﺷ
Bioengineered noncoding RNA agents (BERAs)
شﻮﻣﺎﺧ ياﺮﺑ ،
فﺪﻫ نژ ندﺮﮐ نوﺮﺑ لﺪﻣ رد ،GFP
ﻮﻤﻧ هرﺎﺷا ﯽﺷﻮﻣ و ﯽﻨﺗ ﻦﯾا .د
ﻪﮐ داد نﺎﺸﻧ ﻪﻌﻟﺎﻄﻣ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ مﺮﺳ يراﺪﯾﺎﭘ ﺶﯾاﺰﻓا ﺐﺒﺳ ﺎﻫ
siRNA
لﻮﻠﺳ ﻪﺑ ﺮﮕﺷﻮﻣﺎﺧ نژ ﺪﻣآرﺎﮐ لﺎﻘﺘﻧا ،ﺎﻫ و يرﻮﻣﻮﺗ يﺎﻫ
نآ نﺎﯿﺑ ﺶﻫﺎﮐ ﯽﻣ
ﺪﻧﻮﺷ . يﺮﮕﯾد ﻪﻌﻟﺎﻄﻣ رد 17
ﯽﻟﻮﮑﻟﻮﻣ نزو ﺎﺑ PEI
) ﻦﯿﯾﺎﭘ 600 وﺮﺘﺳا ﺮﮑﻨﯿﻟ ﻪﺳ ﻪﺑ ﻞﺼﺘﻣ (نﻮﺘﻟاد ﯾ
ﺎﺑ ﺐﯿﮐﺮﺗ رد يﺪﯿ
لﺎﻘﺘﻧا ياﺮﺑ موزﻮﭙﯿﻟ هدر ﻪﺑ نژ
ﯽﻟﻮﻠﺳ يﺎﻫ
HeLa, HEK293
و
MCF-7
ﻞﻣﺎﺣﻮﻧﺎﻧ هزاﺪﻧا .ﺖﻓﺮﮔ راﺮﻗ هدﺎﻔﺘﺳا درﻮﻣ ﺎﻫ
300 nm
- 200 ﻦﯿﻤﺨﺗ
لﻮﻠﺳ ﻪﺑ ار نژ ﺪﻧدﻮﺑ ردﺎﻗ ﻪﮐ ﺪﺷ هدز ﺪﻨﻨﮐ ﻞﻘﺘﻨﻣ ﺎﻫ
.
22
) ﺐﺳﺎﻨﻣ هزاﺪﻧا ياراد ﻪﻠﺻﺎﺣ تارذﻮﻧﺎﻧ 104nm
ﻪﺑ ( ﯽﻣ ﺮﻈﻧ ﺪﯿﺳر
.ﺪﻨﺷﺎﺑ ﺐﺳﺎﻨﻣ نژ لﺎﻘﺘﻧا ياﺮﺑ ﯽﻠﭘﻮﭙﯿﻟ
ﺲﮑﻠﭘ ﺎﻫ ﺖﺒﺴﻧ رد 4 /0
= ﺎﺑC/P
ﻪﺑ ﺖﺒﺴﻧ ار ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺶﯾاﺰﻓا يﺮﻤﯿﻠﭘ ﻪﯾﺎﭘ ود ﺮﻫ 25 PEI
ﻪﺑ) نﻮﺘﻟادﻮﻠﯿﮐ ﯽﻣ ﻪﮐ ﺪﻧداد نﺎﺸﻧ (دراﺪﻧﺎﺘﺳا ناﻮﻨﻋ
ﺖﻠﻋ ﻪﺑ ﺪﻧاﻮﺗ ﺶﻫﺎﮐ
ﻪﺑ ﺖﺒﺴﻧ ﺖﯿﻤﺳ هﺪﻧزﺎﺳ داﻮﻣ
هﺪﺸﻧ يزرو ﺖﺳد .ﺪﺷﺎﺑ
:يراﺰﮕﺳﺎﭙﺳ اﯾ
ﻦ ﻘﺤﺗ حﺮﻃ ﻞﺻﺎﺣ ﻪﻟﺎﻘﻣ ﯿ
ﺗﺎﻘ ﯽ ناﻮﻨﻋ ﺖﺤﺗ
"
ﺳرﺮﺑ ﯽ ارﺎﮐ ﯽﯾ ﻤﺳ و ﯿ ﺖ ﯾ ﮏ ﻞﻣﺎﺣ ﯽﻠﭘﻮﭙﯿﻟ ﺲﮑﻠﭘ ﻦﺸﮑﻔﺴﻧاﺮﺗ ﺖﻬﺟ
تﺎﺘﺳوﺮﭘ نﺎﻃﺮﺳ هدر ﺎﻘﻣ و PC3
ﯾ ﻪﺴ ﺎﺑ ﯾ ﮏ رﺎﺠﺗ ﻞﻣﺎﺣ ي
لﻮﻤﻌﻣ
"
ﮑﺷﺰﭘ مﻮﻠﻋ هﺎﮕﺸﻧاد بﻮﺼﻣ ﯽ
لﺎﺳ رد ﺪﻬﺸﻣ 1397
ﺪﮐ ﺎﺑ 960095
ﻣ ﯽ ﺎﻤﺣ ﺎﺑ ﻪﮐ ﺪﺷﺎﺑ ﯾ
ﺖ ﻤﮐ ﯿ ﻪﺘ ﻘﺤﺗ ﯿ تﺎﻘ ﻮﺠﺸﻧاد ﯽﯾ مﻮﻠﻋ هﺎﮕﺸﻧاد
ﮑﺷﺰﭘ ﯽ .ﺖﺳا هﺪﺷ اﺮﺟا ﺪﻬﺸﻣ
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2019;126(2):S111.
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Investigating lipopolymers for gene delivery to prostate cancer cells
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Investigating lipopolymers based on polyethylenimine and nanoliposome for gene delivery to prostate cancer (PC3) cell line
Mohammad Hasan Jafari Najaf Abadi Ph.D.1Ϯ
Saeedeh Askarian Ph.D.2,3Ϯ Reza Kazemi Oskuee Ph.D.1 Bizhan Malaekeh-Nikouei Ph.D.4
Mehdi Rezaee Ph.D.1
Seyed Hamid Aghaee-Bakhtiari Ph.D.1,5*
1- Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
2- Department of Medical Biotechnology, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran.
3- Neuroscience Research Center, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran.
4- Nanotechnology Research Center, Institute of Pharmaceutical Technology, Mashhad University of Medical Sciences, Mashhad, Iran.
5- Bioinformatics Research Group, Mashhad University of Medical Sciences, Mashhad, Iran.
ϮMohammad Hasan Jafari Najaf Abadi and Saeedeh Askarian Contributed as Joint First Author
*Corresponding author: Department of Medical Biotechnology, Third Floor, Faculty of Medicine, Mashhad University of Medical Sciences, Azadi Sq., Mashhad, Iran.
Tel: +98-51-38002300 E-mail: [email protected]
Background: Non-viral Nano carriers such as liposomes and cationic polymers based on engineered properties are regarded in gene delivery field. Although these carriers do not have weaknesses of viral vectors, but they are less efficient than viruses and they still need to be improved as favorable gene delivery carriers. Amongst non-viral carriers, cationic liposomes have been proposed for clinical applications, but limitations such as low nucleic acid transfer and endosome escape and conduction of plasmid to the nucleus have challenged their use in clinical trials. Therefore, the combination of liposomes and cationic polymers for nucleic acid transfer has been considered because this approach makes it possible to use the desirable properties of liposomes and polymers so that it is even suggested for the gene treatment of some diseases such as Parkinson's. In this study, a combination of liposomes and cationic polymers were used for the preparation of lipopolyplexes. This approach allows simultaneous utilizing of the desirable properties of liposomes and polymers.
Methods: This interventional-experimental study was conducted in the medical faculty of Mashhad University of Medical Sciences from April 2017 to February 2018. In this study, PEI-based lipopolyplex with a molecular weight of 25 and 10 kDa and a liposome-to-polymer ratio of 1:1 were combined with plasmid containing the GFP (Green Fluorescent Protein) marker. The physicochemical properties of the synthesized carriers such as size, cytotoxicity and gene transferability in human prostate cancer (PC3) cells were evaluated.
Results: The prepared lipopolyplex were 104 nm in size and all the lipopolyplexes were able to enhance transfection in the C/P=0.4 compared with its basic carriers (PEI and liposomes) alone, while showing less cytotoxicity than not manipulated liposomes. The results of this study suggest synthesized nanoparticles as nanocomposites for gene delivery purposes to different cells and in in-body studies.
Conclusion: The results of this study show that the lipopolyplex constructed from combination of PEI and liposomes can efficiently transfer the gene to the cell, while showing low cytotoxicity and appropriate size at the nano-scale. Therefore, this lipopolymer can be suggested for gene delivery purposes to different cells and in vivo targets.
Keywords: gene transfer techniques, liposomes, PC-3 cells, transfection.
Abstract Received: 21 Feb. 2020 Revised: 25 Feb. 2020 Accepted: 13 Aug. 2020 Available online: 20 Aug. 2020
Original Article Tehran University Medical Journal, August 2020; Vol. 78, No. 5: 284-292
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