fa ارزیابی تغییرات بیان ژن های مسیر میتوکندریایی آپوپتوز و چرخه سلولی در سلول های AsPC-1 تیمار شده با فلورواوراسیل علوم سلولی و مولکولی Cell and Molecular Biology مقاله پژوهشی Original Article <span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:Calibri,sans-serif">&nbsp;<span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">سرطان پانکراس یکی از کشند‌ه&shy;ترین و تهاجمی‌&shy;ترین سرطان&shy;‌ها است؛ فلورواوراسیل باعث توقف در چرخه سلولی و القای آپوپتوز در سلول&shy;های سرطانی می&shy;شود. در مطالعه حاضر اثر<a name="_Hlk103714398"> فلورواوراسیل</a> بر مراحل مختلف چرخه سلولی و بیان ژن&shy;های درگیر در مسیر درونی آپوپتوز در رده سلولی </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">AsPC-1</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> (سرطان پانکراس انسانی)، بررسی شده است. در این پژوهش تجربی-آزمایشگاهی برای بررسی اثر سمیت فلورواوراسیل بر روی تکثیر سلول&shy;های </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">AsPC-1</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> از آزمون </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">MTT</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> استفاده شد؛ نوع مرگ سلولي القا شده و تغییرات در چرخه سلولي به روش فلوسايتومتري بررسي شد؛ تغییرات در سطح بیان ژن&shy;‌های (</span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">BAX</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">، </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">Bcl-2</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">، </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">APAF-1</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">، </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">Caspase-3</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">، </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">Caspase-9</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">، </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">p53</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> و </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">p21</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">) با تکنیک </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">Real-time PCR</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> ارزیابی شد. داده‌&shy;های کمی حاصل در سطح معنی‌&shy;داری (</span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">p< 0.05</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">) تحلیل گردید. یافته&shy;‌های حاصل از آزمون </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">MTT</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> نشان داد که فلورواوراسیل به</span></span></span><span lang="FA" style="font-size:10.0pt"><span style="letter-spacing:-.3pt">&shy;‌</span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">صورت وابسته به غلظت باعث کاهش تکثیر سلول&shy;های </span></span></span><a name="_Hlk103721695"><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">AsPC-1</span></span></span></a><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> می&shy;‌شود، نتایج آنالیز فلوسایتومتری نشان دهنده افزایش درصد سلول&shy;‌های آپوپتوتیک در سلول&shy;‌های تحت تیمار بود؛ فلورواوراسیل در سلول&shy;های </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">AsPC-1</span></span></span> <span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">سبب توقف چرخه سلولی در مرحله </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">S</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> و کاهش جمعیت سلولی در فاز </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">G1</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> شد. بررسی نتایج </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">Real-time PCR</span></span></span> <span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">در سلول&shy;‌های تیمار شده، نشان دهنده افزایش بیان ژن&shy;های مسیر میتوکندریایی آپوپتوز و همچنین ژن&shy;های موثر در تنظیم چرخه سلولی بود. فلورواوراسیل باعث کاهش تکثیر سلولی و القای آپوپتوز از طریق افزایش بیان ژن&shy;های دخیل در مسیر درونی آپوپتوز در سلول&shy;های </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">AsPC-1</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt"> می&shy;‌شود، همچنین فلورواوراسیل با تنظیم ژن&shy;های (</span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">p53</span></span></span> <span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">و </span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">p21</span></span></span><span lang="FA" style="font-size:10.0pt"><span b="" nazanin="" style="font-family:"><span style="letter-spacing:-.3pt">) سبب توقف چرخه سلولی در این سلول‌ها می&shy;‌شود.</span></span></span><span dir="LTR" lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt"></span></span></span></span></span></span></span></span></span><br /> <span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:Calibri,sans-serif"><b><span dir="LTR" lang="EN-CA" style="font-size:5.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt"></span></span></span></b></span></span></span></span></span></span><br /> &nbsp; <span style="font-size:11pt"><span style="line-height:normal"><span style="font-family:Calibri,sans-serif"><span lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">Pancreatic cancer is one of the most deadly and aggressive cancers; Fluorouracil induces apoptosis and cell cycle arrest in cancer cells. In the present study; the effect of Fluorouracil on different stages of the cell cycle and the expression of genes involved in the internal pathway of apoptosis in the AsPC-1 cell line (human pancreatic cancer) were investigated. In order to do so, MTT assay was used to evaluate the cytotoxic effect of Fluorouracil on AsPC-1 cell proliferation; The type of induced cell death and cell cycle changes were investigated by flow cytometry; changes in the expression level of genes (BAX, Bcl-2, APAF-1, Caspase-3, Caspase-9, p53, p21) were examined by Real-time PCR. Quantitative data were analyzed at the significant level of (p<0.05). </span></span></span><span lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">The MTT assay results showed that Fluorouracil decreased AsPC-1 cell proliferation in a concentration-dependent manner. The results of </span></span></span><span lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">flow cytometry</span></span></span><span lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt"> analysis showed that increased percentage of apoptotic cells in the treated cells; Fluorouracil induces S phase cell cycle arrest in AsPC-1 cells and reduced distribution in the G1 phase. The Real-time PCR results in treated cells showed an increase in the expression of genes in the mitochondrial apoptotic pathway as well as genes effective in regulating the cell cycle. </span></span></span><span lang="EN-CA" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.3pt">Fluorouracil reduces cell proliferation and induces apoptosis by increasing the expression of genes involved in the Intrinsic apoptotic pathway in AsPC-1 cells; Fluorouracil also caused cell cycle arrest in these cells by regulating the (p53, p21) genes.</span></span></span></span></span></span><br /> &nbsp; تکثیر سلولی, تکنیک Real-time PCR, سرطان پانکراس, فلوسایتومتری cell proliferation, flow cytometry, pancreatic cancer, real-time PCR 99 107 http://nbr.khu.ac.ir/browse.php?a_code=A-10-1472-2&slc_lang=fa&sid=1 Negar Khorasani نگار خراسانی negar.khorasani86@gmail.com 100319475328460010275 100319475328460010275 No Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran گروه زیست شناسی، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران Javad Baharara جواد بهارآرا baharara@mshdiau.ac.ir 100319475328460010276 100319475328460010276 Yes Department of Biology & Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran گروه زیست شناسی و مرکز تحقیقات بیولوژی کاربردی تکوین جانوری، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران Khadijeh Nejad Shahrokhabadi خدیجه نژاد شاهرخ آبادی shahrokhabady@yahoo.com 100319475328460010277 100319475328460010277 No Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran گروه زیست شناسی، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران