ارزيابي تأثير بيان افزايشي هومئودومين پروتئين TGIF2LX بر روي تغييرات بيان miRNA‌هاي انكوژن در رده‌ي سلولي سرطان روده بزرگ SW1116

The Effect of Overexpression of Homeodomain Protein TGIF2LX on the Expression of Oncogenic miRNAs in Colorectal Cancer Cell Line SW1116

ﭘﺮوﺗﺌﯿﻦ اﻟﻘﺎ ﺷﻮﻧﺪه ﺗﻮﺳﻂ ﻓﺎﮐﺘﻮر رﺷﺪ ﺗﺮاﻧﺴﻔﻮرم ﮐﻨﻨﺪه ﺑﺘﺎ ﻣﺮﺗﺒﻂ ﺑﺎ TGIF2LX) X( ﺑﻪ ﻋﻨﻮان ﯾﮏ ﻫﻮﻣﺌﻮدوﻣﯿﻦ ﭘﺮوﺗﺌﯿﻦ و ﮐﻮرﭘﺮﺳﻮر ﻓﺎﮐﺘﻮر رﺷﺪ ﺗﺮاﻧﺴﻔﻮرم ﮐﻨﻨﺪه ﺑﺘﺎ ) -TGF (، ﺗﮑﺜﯿﺮ ﺑﺮﺧﯽ ﺳﻠﻮل ﻫﺎي ﺳﺮﻃﺎﻧﯽ از ﺟﻤﻠﻪ ﺳﺮﻃﺎن روده ﺑﺰرگ را ﺑﻪ واﺳﻄﻪ ي ﺑﺮﺧﯽ ﻣﺴﯿﺮﻫﺎي ﭘﯿﺎم رﺳﺎﻧﯽ ﺗﻨﻈﯿﻢ ﻣﯽ ﮐﻨﺪ. NA ﻫﺎي ﻏﯿﺮﮐﺪﮐﻨﻨﺪه ﮐﻮﭼﮏ )(micro RNA; miRNA ﺑﻪ ﻋﻨﻮان ﺗﻨﻈﯿﻢ ﮐﻨﻨﺪه ﻫﺎي ﻣﻮﻟﮑﻮﻟﯽ ﺳﺮﻃﺎن روده ﺑﺰرگ ﺷﻨﺎﺧﺘﻪ ﻣﯽ ﺷﻮﻧﺪ ﮐﻪ در ﻓﺮآﯾﻨﺪﻫﺎي رﺷﺪ، ﺗﮑﺜﯿﺮ، ﺗﻤﺎﯾﺰ و آﭘﻮﭘﺘﻮز ﺳﻠﻮﻟﯽ ﻧﻘﺶ دارﻧﺪ. ﻫﺪف از اﯾﻦ ﻣﻄﺎﻟﻌﻪ ارزﯾﺎﺑﯽ اﻫﻤﯿﺖ ﺑﯿﻮﻟﻮژﯾﮑﯽ ﭘﺮوﺗﺌﯿﻦTGIF2LX و ﺗﺄﺛﯿﺮ آن ﺑﺮ ﻣﯿﺰان ﺑﯿﺎن miRNA ﻫﺎي اﻧﮑﻮژن miR-20a ، miR-34a و 21-miR در ﺳﻠﻮل ﻫﺎي رده ﺳﺮﻃﺎﻧﯽ روده ﺑﺰرگ SW1116 ﺑﻮد. ﻣﻮاد و روش ﻫﺎ: رده ﺳﻠﻮﻟﯽ SW1116 اﻧﺴﺎن و رده ﺳﻠﻮﻟﯽ ﺗﺮاﻧﺴﻔﮑﺖ ﺷﺪه ﺑﺎ cDNA ﮐﺪﮐﻨﻨﺪه ژن TGIF2LX و ﺑﯿﺎن ﮐﻨﻨﺪه اﻓﺰاﯾﺸﯽ اﯾﻦ ﻓﺎﮐﺘﻮر، در ﻣﺤﯿﻂ ﮐﺸﺖ RPMI 1640 در ﺷﺮاﯾﻂ ﻣﻨﺎﺳﺐ ﮐﺸﺖ ﺷﺪﻧﺪ. ﺑﺮاي ارزﯾﺎﺑﯽ ﻣﯿﺰان ﺣﯿﺎت ﺳﻠﻮل ﻫﺎ در in vitro از آزﻣﻮن MTT اﺳﺘﻔﺎده ﺷﺪ. ﭘﺲ از اﺳﺘﺨﺮاجRNA از ﻫﺮ دو ﮔﺮوه ﺳﻠﻮﻟﯽ و ﺳﻨﺘﺰ cDNA ، ﺗﺠﺰﯾﻪ و ﺗﺤﻠﯿﻞ ﺑﯿﺎن miRNA ﻫﺎ ﺑﺎ اﺳﺘﻔﺎده از ﺗﮑﻨﯿﮏ qRT-PCR اﻧﺠﺎم ﺷﺪ. ﻧﺘﺎﯾﺞ: ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﺑﯿﺎن اﻓﺰاﯾﺸﯽTGIF2LX ﻣﯽ ﺗﻮاﻧﺪ ﺗﮑﺜﯿﺮ رده ﺳﻠﻮﻟﯽ SW1116 را ﮐﺎﻫﺶ دﻫﺪ. ﺗﺠﺰﯾﻪ و ﺗﺤﻠﯿﻞ ﺑﯿﺎن ژن ﻧﺸﺎن داد ﺑﯿﺎن اﻓﺰاﯾﺸﯽ TGIF2LX ﻣﯽ ﺗﻮاﻧﺪ ﺳﻄﺢ ﺑﯿﺎن 21-miR را ﺑﻪ ﻃﻮر ﻣﻌﻨﺎداري ﮐﺎﻫﺶ دﻫﺪ )P=0/026 (. ﺑﺎ اﯾﻦ ﺣﺎل ﺳﻄﺢ ﺑﯿﺎن P=0/52) miR-34a ( و miR-20a )P=0/48 ( در ﺳﻠﻮل ﻫﺎي SW1116 ﺗﺮاﻧﺴﻔﮑﺖ ﺷﺪه ﺑﺎ TGIF2 LX در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﺳﻠﻮ ل ﻫﺎي دﺳﺘﮑﺎري ﻧﺸﺪه ﺗﻐﯿﯿﺮ ﻣﻌﻨﺎداري ﻧﺸﺎن ﻧﺪاد. ﻧﺘﯿﺠﻪ ﮔﯿﺮي: ﯾﺎﻓﺘﻪ ﻫﺎي ﻣﺎ ﺷﻮاﻫﺪي از ﻣﮑﺎﻧﯿﺴﻢ ﻫﺎي ﻣﻮﻟﮑﻮﻟﯽ را اراﯾﻪ داد ﮐﻪ ﻫﻮﻣﺌﻮدوﻣﯿﻦ ﭘﺮوﺗﺌﯿﻦ TGIF2LX ﻣﯽ ﺗﻮاﻧﺪ ﺑﺎ ﮐﺎﻫﺶ ﺑﯿﺎن 21-miR ﺑﻪ ﻋﻨﻮان ﺳﺮﮐﻮب ﮐﻨﻨﺪه ﺗﻮﻣﻮر در ﺳﻠﻮل ﻫﺎي ﺳﺮﻃﺎن روده ﺑﺰرگ ﻋﻤﻞ ﮐﻨﺪ. ﺑﻨﺎﺑﺮاﯾﻦ، ﺑﻪ ﻃﻮر ﺑﺎﻟﻘﻮه ﻣﯽ ﺗﻮان اﯾﻦ ﭘﺮوﺗﺌﯿﻦ را ﺑﻪ ﻋﻨﻮان ﯾﮏ ﮔﺰﯾﻨﻪ اﻣﯿﺪوارﮐﻨﻨﺪه ﺑﺮاي اﺳﺘﺮاﺗﮋي ﻫﺎي درﻣﺎﻧﯽ ﻣﺒﺘﻨﯽ ﺑﺮ ژن در اﯾﻦ ﺳﺮﻃﺎن ﻣﺪﻧﻈﺮ ﻗﺮار داد.

Transforming growth factor-beta-induced factor X-linked (TGIF2LX) as a homeodomain protein and TGF-β corepressor, could regulate proliferation of some cancer cells including colorectal cancer by some signaling pathways. Small non-coding RNAs (microRNA; miRNA) are known as molecular regulators of colorectal cancer that are involved in the processes of cell growth, proliferation, differentiation and apoptosis. The aim of this study was to evaluate the biological significance of TGIF2LX protein and its effect on the expression of oncogenic miRNAs miR-34a, miR-20a and miR-21 in colorectal cancer cells SW1116. Methods: Human SW1116 cell line and cell line transfected with cDNA encoding TGIF2LX gene were cultured in RPMI 1640 medium under appropriate conditions. MTT assay was used to assess cell viability in vitro. After RNA extraction from all cell groups and cDNA synthesis, miRNA expression analysis was performed using qReal-time PCR technique. Results: The results showed that the increased expression of TGIF2LX could reduce the proliferation of SW1116 cell line. Gene expression analysis showed that increased expression of TGIF2LX could significantly reduce the expression level of miR-21 (P<0.038). However, the expression levels of miR-34a and miR-20a in SW1116 cells transfected with TGIF2LX did not show a significant change compared to non-transfected cells (P>0.05). Conclusion: Our findings provide evidence of molecular mechanisms that the homeodomain protein TGIF2LX can act as a tumor suppressor in colorectal cancer cells by reducing miR-21 expression. Therefore, this protein can potentially be considered as a promising option for gene-based therapeutic strategies in this cancer.