مطالعه اثر غلظت هاي مختلف عصاره زعفران بر الگوي بياني برخي از ژن هاي كنترل كننده مسير خودبازآفريني ( Self-Renewal) سلول در رده سلول سرطاني AGS (آدنوكارسينوماي معده)

The Survey of Saffron Extract Effects on Expressional Pattern of Some Self-renewal genes in AGS (gastric adenocarcinoma) Tumor Cell Line

ﻣﻘﺪﻣﻪ: اﺳﺘﻔﺎده از ﺗﺮﮐﯿﺒﺎت ﮔﯿﺎﻫﯽ ﺑﻪﻋﻨﻮان ﺳﺮﮐﻮبﮐﻨﻨﺪه ﺳﺮﻃﺎن، اﻣﺮوزه اﻫﻤﯿﺖ زﯾـﺎدي ﭘﯿـﺪا ﮐـﺮده اﺳـﺖ. ﻫـﺪف از ﻣﻄﺎﻟﻌـﻪ ﺣﺎﺿـﺮ ﺑﺮرﺳـﯽ ﺗـﺄﺛﯿﺮ ﻏﻠﻈـﺖ ﻫـﺎي ﻣﺨﺘﻠـﻒ ﻋﺼـﺎره زﻋﻔـﺮان rocus sativus ﺑـﺮ ﺑﯿـﺎن ژن ﻫـﺎي NANOG ،SOX2 ،KLF ،OCT4 و Nucleostemin در رده ﺳﻠﻮﻟﯽ Gastric Adenocarcinoma AGS بود. ﻣﻮاد و روشﻫﺎ: رده ﺳﻠﻮل ﺳﺮﻃﺎﻧﯽ آدﻧﻮﮐﺎرﺳﯿﻨﻮم ﻣﻌﺪه (AGS) از ﺑﺎﻧﮏ ﺳﻠﻮﻟﯽ اﻧﺴﺘﯿﺘﻮ ﭘﺎﺳﺘﻮر اﯾﺮان ﺗﻬﯿﻪ و در ﻣﺤﯿﻂ ﮐﺸﺖ ﮐﺸﺖ داده ﺷﺪ. ﭘﺲ از ﭼﻨﺪ ﭘﺎﺳﺎژ، اﻃﻤﯿﻨﺎن از ﺻﺤﺖ ﺳﻠﻮلﻫﺎ، ﺳﻠﻮلﻫﺎ در دو ﮔﺮوه ﺗﺴـﺖ و ﮐﻨﺘـﺮل ﺑـﺎ ﺷـﺮاﯾﻂ ﯾﮑﺴـﺎن ﮐﺸـﺖ ﺷﺪﻧﺪ. ﭘﺲ از رﺳﯿﺪن ﺗﺮاﮐﻢ ﺳﻠﻮﻟﯽ ﺑﻪ 60-70 درﺻﺪ، ﮔﺮوه آزﻣﺎﯾﺶ در ﻣﻌـﺮض ﻏﻠﻈـﺖ ﻫـﺎي 20، 40 و 100 ﻣﯿﮑﺮوﮔـﺮم در ﻣﯿﻠـﯽ ﻟﯿﺘـﺮ ﻋﺼﺎره زﻋﻔﺮان ﻗﺮار ﮔﺮﻓﺘﻪ و ﭘﺲ از ﮔﺬﺷﺖ ﺑﺎزهﻫﺎي زﻣﺎﻧﯽ 24، 48 و 72 ﺳﺎﻋﺖ، ﺳﻠﻮلﻫﺎ ﺟـﺪا ﺷـﺪه، ﭘـﺲ از ﺗﺨﻠـﯿﺺ Total RNA و ﺳﻨﺘﺰ cDNA، ﻣﯿﺰان ﺑﯿﺎن ژنﻫﺎي ﻣﻮرد ﻧﻈﺮ و ﻣﯿﺰان ﺗﻐﯿﯿﺮات ﺑﯿﺎﻧﯽ اﯾﻦ ژنﻫﺎ در دو ﮔﺮوه ﺗﺴﺖ و ﮐﻨﺘﺮل ﺗﻌﯿﯿﻦ ﮔﺮدﯾﺪ. ﯾﺎﻓﺘﻪﻫﺎ: ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﻋﺼﺎره زﻋﻔﺮان ﺑﺎﻋﺚ ﮐﺎﻫﺶ ﺑﯿﺎن ژنﻫﺎي NANOG ،SOX2 ،KLF4 ،OCT4 و Nucleosteminمي شود. ﻧﺘﯿﺠﻪﮔﯿﺮي: ﻧﺘﺎﯾﺞ اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﻧﺸﺎن داد ﮐﻪ ﻋﺼﺎره زﻋﻔﺮان ﻣﯽﺗﻮاﻧﺪ ﺑﻪ ﻋﻨﻮان ﯾﮏ ﻋﺎﻣﻞ ﻣﻬﺎرﮐﻨﻨﺪه ﺗﻘﺴﯿﻤﺎت ﺳـﻠﻮﻟﯽ، در ﭘﯿﺸـﮕﯿﺮي از ﺳﺮﻃﺎن ﮐﻪ ﺑﻪ واﺳﻄﻪ ﺗﻘﺴﯿﻤﺎت ﮐﻨﺘﺮل ﻧﺸﺪه ﺳﻠﻮﻟﯽ اﯾﺠﺎد ﻣﯽﺷﻮد، ﻣﻮرد ﺗﻮﺟﻪ ﻗﺮار ﮔﯿﺮد. ﻫﻤﭽﻨﯿﻦ اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﻣﯽﺗﻮاﻧـﺪ ﺑﺴـﺘﺮي ﺑـﺮاي ﻣﻄﺎﻟﻌﺎت ﮔﺴﺘﺮدهﺗﺮ در ﺧﺼﻮص ﺗﺄﺛﯿﺮ ﺿﺪ ﺳﺮﻃﺎﻧﯽ اﯾﻦ ﺗﺮﮐﯿﺐ ﮔﯿﺎﻫﯽ ﺑﺎﺷﺪ.

Introduction: The aim of the present study was to investigate the effect of different concentrations of saffron (Crocus sativus) on the expression of OCT4, KLF, SOX2, NANOG and Nucleostemin genes in AGS (gastric cancer) cell line. Materials and Methods: Gastric adenocarcinoma cell line (AGS) was prepared from the cell bank of Iran Pasteur Institute and were cultured in RPMI 1640 medium. The cells were then cultured in both test and control groups under similar conditions. At confluency of 60-70%, the test group was treated with saffron essence at concentrations of 20, 40 and 100 μg/ml for 24, 48 and 72 hours. Total RNA was extracted prior to cDNA preparation. The expression of target genes in both test and control groups were determined. Results: The results showed that the saffron extract decreased expression of OCT4, KLF, SOX2, NANOG and Nucleostemin genes. Conclusion: The results suggested that saffron can be considered as an anti-cancer agent due to its role in inhibition of cell division. This study could be the basis for further studies on the anti-cancer property of herbal products.