Mobalegh Naseri, Mahshid

Simorgh Research Repository

      ﭘﺰﺷਔﯽ ໊ඟﻣﺎن دا஺ه ﻋ࢖ﻮم  داروﺳﺎزيداﻧﺸﻜﺪه  ﺳﻢ ﺷﻨﺎﺳﻲ ﭘﺰﺷﻜﻲﭘﺎﻳﺎن ﻧﺎﻣﻪ ﻣﻘﻄﻊ ﻛﺎرﺷﻨﺎﺳﻲ ارﺷﺪ   ﻋﻨﻮان:رده  ﺑﺮﺣﺎوي ﻛﻮرﻛﻮﻣﻴﻦ detegrat-ydobiffA 2REHZ  ﺑﺮرﺳﻲ و ﻣﻘﺎﻳﺴﻪ اﺛﺮ ﻟﻴﭙﻮزوﻣﻬﺎي  3-RBKS و  7-FCMﻫﺎي ﺳﻠﻮﻟﻲ ﺳﺮﻃﺎن ﭘﺴﺘﺎن      ﺪ ﻣﺒﻠﻎ ﻧﺎﺻﺮيﻴﻣﻬﺸ ﺗﻮﺳﻂ:  ﻋﻠﻲ ﻣﺎﻧﺪﮔﺎري ﺑﺎ ﻣﻜﺎن دﻛﺘﺮ راﻫﻨﻤﺎ: ﺗﻴﺪﺎاﺳ  دﻛﺘﺮ ﻃﻮﺑﻲ اﺳﻼﻣﻲ ﻧﮋاد  دﻛﺘﺮ ﻣﺠﻴﺪ زﻳﻨﻠﻲ  ﺑﻬﺰاد ﺑﻬﻨﺎم دﻛﺘﺮ ﻣﺸﺎور: ﺗﻴﺪﺎاﺳ دﻛﺘﺮ ﻋﺒﺎس ﭘﺮداﺧﺘﻲ 7931- 89 ﺳﺎل ﺗﺤﺼﻴﻠﻲ:     ﭼﻜﻴﺪه از ﺑﺴﻴﺎري در. اﺳﺖ ﺷﺪه ﺷﻨﺎﺧﺘﻪ ﺳﺮﻃﺎن درﻣﺎن ﻣﺆﺛﺮ در ﮔﻴﺎﻫﻲ داروي ﻳﻚ ﻋﻨﻮان ﺑﻪ ﻛﻮرﻛﻮﻣﻴﻦ :و اﻫﺪاف ﻣﻘﺪﻣﻪ ﺳﺮﻃﺎن درﻣﺎن ﻫﺪﻓﻤﻨﺪ درﻛﻪ  ﺷﻮد ﻣﻲ ﺑﻴﺎن ﺑﻴﺶ از ﺣﺪ )2 REH( 2 ﻧﻮع اﻧﺴﺎﻧﻲ اﭘﻴﺪرﻣﻲ رﺷﺪ ﻓﺎﻛﺘﻮر ﮔﻴﺮﻧﺪه ﻧﻮع ﻫﺎ، ﺳﺮﻃﺎن ﺑﺮ  )ydobiffA( اﻓﻲ ﺑﺎدي ﻛﺎﻧﮋوﮔﻪ ﺷﺪه ﺑﺎ ﻫﺎي ﻟﻴﭙﻮزوم در ﺷﺪه ﮔﺬاريﺑﺎر ﻛﻮرﻛﻮﻣﻴﻦ اﺛﺮ ﻣﻄﺎﻟﻌﻪ اﻳﻦ در .اﺳﺖ ﻣﻬﻢ ﺑﺴﻴﺎر  ﺑﺮرﺳﻲ ﺷﺪه اﺳﺖ. دارﻧﺪ 2REHﻛﻪ ﺑﻴﺎن ﻣﺘﻔﺎوﺗﻲ از ﮔﻴﺮﻧﺪه ي  7-FCM ،3RBKS ﭘﺴﺘﺎن ﺳﺮﻃﺎن ﻫﺎي ﺳﻠﻮلﻫﻴﺪرﻳﺸﻦ و ﺑﻪ روش ﻟﻴﭙﻴﺪ ﻓﻴﻠﻢ  2:3  )w/w(از ﻟﻴﺴﻴﺘﻴﻦ ﺳﻮﻳﺎ و ﻓﺴﻔﻮﻟﻴﭙﻴﺪ ﺑﺎ ﻧﺴﺒﺖﻟﻴﭙﻮزوم ﻫﺎ و اﻳﻤﻮﻧﻮﻟﻴﭙﻮزوم ﻫﺎ  :ﻫﺎ روشﺗﺤﺖ  1 lm/gmﺗﻬﻴﻪ ﺷﺪﻧﺪ، ﺳﭙﺲ ﺗﻮﺳﻂ ﻟﻴﻨﻜﺮ ﻣﺎﻟﻤﺎﻳﺪ ﺑﻪ اﻓﻲ ﺑﺎدي ﻣﺘﺼﻞ ﺷﺪه و ﭘﺲ از ﻫﻴﺪراﺗﻪ ﺷﺪن ﺑﺎ دارو ﺑﺎ ﻏﻠﻈﺖ ﮔﺮاد و ﻫﻤﺰدن ﻣﺪاوم در ﺣﻤﺎم ﻓﺮاﺻﻮت ﻗﺮار داده ﺷﺪﻧﺪ، ﺳﭙﺲ ﺑﻪ روش ژل ﻛﺮوﻣﺎﺗﻮﮔﺮاﻓﻲ ﺧﺎﻟﺺ  درﺟﻪ ي ﺳﺎﻧﺘﻲ 55دﻣﺎي ﺑﺮرﺳﻲ ﺷﺪﻧﺪ. ﻣﻴﺰان  )MFA( ﻲاﺗﻤاز ﻣﻴﻜﺮوﺳﻜﻮپ ﻫﺎ ﺑﺎ اﺳﺘﻔﺎده  ﻧﻮﻟﻴﭙﻮزوماﻳﻤﻮو  ﻫﺎ ﻟﻴﭙﻮزوم اﻧﺪازه ذره ايﺳﺎزي ﺷﺪﻧﺪ. ﺳﻠﻮﻟﻲ ذرات ﺳﻤﻴﺖ  ﮔﻴﺮي ﺷﺪﻧﺪ.اﻧﺪازه ﺑﺎ اﺳﺘﻔﺎده از دﺳﺘﮕﺎه اﺳﭙﻜﺘﺮوﻓﺘﻮﻣﺘﺮي داروي ﺑﺎرﮔﺬاري ﺷﺪه در ذرات ﺳﺎﺧﺘﻪ ﺷﺪه  روش ﺑﺎ ﺲﺳﭙ و ﺑﺮرﺳﻲ ﺷﺪﻧﺪ TTM روش از اﺳﺘﻔﺎده ﺑﺎ 7-FCM ،3RBKSﺑﺮ روي رده ﻫﺎي ﺳﺮﻃﺎﻧﻲ  ﺷﺪه ﺳﺎﺧﺘﻪ   .ﮔﺮﻓﺖ ﻗﺮار ﺑﺮرﺳﻲ ﻣﻮرد اﺣﺘﻤﺎﻟﻲ آن ﻣﺴﻴﺮ و ﭘﺘﻮزﻮﭘآ روﻧﺪ ﻓﻠﻮﺳﺎﻳﺘﻮﻣﺘﺮي، ﭘﺬﻳﺮي ﭘﺨﺶ ﺿﺮﻳﺐ و ﻧﺎﻧﻮﻣﺘﺮ 441 ﻛﻮرﻛﻮﻣﻴﻦ ﺣﺎوي ﻫﺎي ﻟﻴﭙﻮزوم ﻗﻄﺮاﻧﺠﺎم ﺷﺪه ﺑﺮ ﻃﺒﻖ اﻧﺪازه ﮔﻴﺮي ﻫﺎي  ﻳﺎﻓﺘﻪ ﻫﺎ: ﻣﻴﻜﺮوﮔﺮاف  ﻫﻤﭽﻨﻴﻦ. اﺳﺖ ﺑﻮده 0/991 ﭘﺬﻳﺮي ﭘﺨﺶ ﺿﺮﻳﺐ و ﻧﺎﻧﻮﻣﺘﺮ 751 اﻳﻤﻮﻟﻴﭙﻮزوم ﻗﻄﺮ. اﺳﺖ ﺑﻮده 0/8410 ﻫﺎ آنﻃﺒﻖ ﻧﺘﺎﻳﺞ ﺑﻪ  .ﺑﺎﺷﻨﺪ ﻣﻲ ﺷﻜﻞ ﻛﺮوي و ﺻﺎف ﺳﻄﺤﻲ داراي ﻫﺎ ﻟﻴﭙﻮزوم ﻛﻪ دﻫﺪ ﻣﻲ ﻧﺸﺎن اﺗﻤﻲ ﻣﻴﻜﺮوﺳﻜﻮپ ﺑﺎ ﺷﺪه ﺗﻬﻴﻪﺑﻴﺸﺘﺮي ﻧﺴﺒﺖ ﺑﻪ ﻛﻮرﻛﻮﻣﻴﻦ ﺧﺎﻟﺺ ﺑﺮ ﺳﻠﻮل ﻫﺎي ﺳﺮﻃﺎن  اتﻫﺎ اﺛﺮدر اﻳﻤﻮﻧﻮﻟﻴﭙﻮزوم  ﺑﺎرﮔﺰاري ﺷﺪهدﺳﺖ آﻣﺪه ﻛﻮرﻛﻮﻣﻴﻦ ﻧﺎﺘﺴﭘﻲ MCF-7 و SKBR3 ﻧ نﺎﺸﺖﺳا هداد ،ﺎﻣا  ﺮﺛا ناﺰﻴﻣوراد يور ﺮﺑ لﻮﻠﺳ يﺎﻫ زا توﺎﻔﺘﻣ ﺢﺷﺮﺗ ناﺰﻴﻣ ﺎﺑ HER2 ًﺎﺒﻳﺮﻘﺗ نﺎﺴﻜﻳ  هدﻮﺑﺖﺳا.   :يﺮﻴﮔ ﻪﺠﻴﺘﻧ ﺮﺑ سﺎﺳا ﺞﻳﺎﺘﻧ ﺑ ﻪﺳد ﻖﻴﻘﺤﺗ زا هﺪﻣآ ﺖ رﻮﻛﺬﻣ ﻪﻧﺎﻣﺎﺳ ﺪﻳﺪﺟ ﻲﻳوراد دﺮﻜﻠﻤﻋ دﻮﺒﻬﺑ ﺐﺟﻮﻣ هﺪﺷ ﻲﺣاﺮﻃﻲﻣ ﻦﻴﻣﻮﻛرﻮﻛ ﺷ يﺎﻘﻟا ﻖﻳﺮﻃ زا ار دﻮﺧ ﺮﺛا و دﻮﻮﭘآﻣ زا و زﻮﺘﭘ زﺎﭙﺳﺎﻛ ﺮﻴﺴ3/7  لﺎﻤﻋاﻲﻣ ﺪﻨﻛ.  :هژاو ﺪﻴﻠﻛ ﻦﻴﻣﻮﻛرﻮﻛ، HER2، ،موزﻮﭙﻴﻟﻮﻧﻮﻤﻳا نﺎﻃﺮﺳ ،نﺎﺘﺴﭘ نﺎﻣرد ﺪﻨﻤﻓﺪﻫ  Abstract Introduction and Objectives: Curcumin is known as an effective herbal medicine for the treatment of cancer. In many cancers, Human Epidermal Growth Factor Receptor type ٢ (HER ٢ )is over-expressed, which is very important in targeted cancer treatment. In this study, the effect of curcumin loaded on affibody conjugated liposomes on SKBR٣, MCF-٧ breast cancer cell lines that expressing different expressions of HER٢ receptors was investigated. Methods: Liposomes and immunoliposomes were prepared from soybeans lecetin and phospholipids at a ratio of ٣: ٢ and in a lipid film hydration method. They were then bonded to the affibody by Malemaid linker and hydrated with the drug at a concentration of ١ mg / ml on Temperature of ٥٥ ° C and stirered in ultrasonic bath, then purified by gel chromatography. Particle size of liposomes and immunoliposomes were evaluated using atomic microscopy (AFM). The amount of drug loaded in the particles was measured using spectrophotometric. The cell cytotoxicity of the particles on the SKBR٣, MCF-٧ cancer strains was investigated using MTT method, and then by flow cytometry, apoptosis and its possible pathway were investigated. Results: According to the measurements, the diameter of the liposomes containing curcumin was ١٤٤٫٢ nm, and their diffusion coefficient was ٠٫١٤٨٨. Imoliposomes diameter was ١٥٥.٧nm and the diffusion coefficient was ٠٫١٩٩. The micrograph prepared with an atomic microscope shows that the liposomes have a smooth and spherical surface. According to the results, curcumin loaded on immunoliposome has been shown to be more effective than pure curcumin on SKBR٣ and MCF-٧ breast cancer cells, but the effect of the drug on cells with different levels of HER٢ secretion is almost the same. Conclusion: Based on the results of this research, the newly designed drug system improves the function of curcumin and its effect is induced by induction of apoptosis from the caspase ٣/٧ pathway. Keywords: Curcumine, HER2, Immunoliposome, Breast cancer, targeted therapy     Kerman University of Medical Sciences Faculty of Pharmacology and Toxicology In Partial Fulfillment of the Requirements for the Degree Medical Toxicology M.Sc.  Title: Investigation and comparison of ZHER2 Affibody-targeted curcumine liposomes on breast tumor cells MCF-7 and SKBR-3  By: Mahshid Moballegh Nasery  Supervisors: Dr. Ali Mandegary Bamakan Dr. Touba Eslaminejad Dr. Majid Zeinali  Advisors: Dr. Behzad Behnam Dr. Abbas Pardakhti Year: 2018-19  

Investigation and comparison of ZHER2 Affibody-targeted curcumine liposomes on breast tumor cells MCF-7 and SKBR-3

http://eprints.kmu.ac.ir/29490/1/6883.pdf

Investigation and comparison of ZHER2 Affibody-targeted curcumine liposomes on breast tumor cells MCF-7 and SKBR-3

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