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Iranian Biomedical Journal، جلد ۲۰، شماره ۳، صفحات ۱۴۵-۱۵۱
عنوان فارسی Suppressive Effect of Constructed shRNAs against Apollon Induces Apoptosis and Growth Inhibition of Hella Cell Line
چکیده فارسی مقاله
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عنوان انگلیسی Suppressive Effect of Constructed shRNAs against Apollon Induces Apoptosis and Growth Inhibition in the HeLa Cell Line
چکیده انگلیسی مقاله Background: Cervical cancer is the second most common female cancer worldwide. Inhibitors of apoptosis proteins (IAPs) block apoptosis; therefore, therapeutic strategies targeting IAPs have attracted the interest of researchers in recent years. Apollon, a member of IAPs, inhibits apoptosis and cell death. RNA interference is a pathway in which small interfering RNA (siRNA) or shRNA (short hairpin RNA) inactivates the expression of target genes. The purpose of this study was to determine the effect of constructed shRNAs on apoptosis and growth inhibition through the suppression of apollon mRNA in HeLa cell line. Methods: Three shRNAs with binding ability to three different target sites of the first region of apollon gene were designed and cloned in pRNAin-H1.2/Neo vector. shRNA plasmids were then transfected in HeLa cells using electroporation. Down-regulation effects of apollon and the viability of HeLa cells were analyzed by RT-PCR, lactate dehydrogenase assay, and MTT assay, respectively. Also, the induction and morphological markers of apoptosis were evaluated by caspase assay and immunocytochemistry method. Results: The expression of shRNA in HeLa cells caused a significant decrease in the level of apollon mRNA1. In addition, shRNA1 effectively increased the mRNA level of Smac (as the antagonist of apollon), reduced the viability of HeLa cells and exhibited immunocytochemical apoptotic markers in this cell line. Conclusion: Apollon gene silencing can induce apoptosis and growth impairment in HeLa cells. In this regard, apollon can be considered a candidate therapeutic target in HeLa cells as a positive human papillomavirus cancer cell line.
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نویسندگان مقاله سعیده میلانی | saeideh milani


مژگان بنده پور | mojgan bandehpour


زهره شریفی | zohreh sharifi


بهرام کاظمی | bahram kazemi


نشانی اینترنتی http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-591&slc_lang=en&sid=en
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کد مقاله (doi)
زبان مقاله منتشر شده fa
موضوعات مقاله منتشر شده Molecular Microbiology
نوع مقاله منتشر شده مقاله کامل
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