fa تنظیم افزایشی miR-155 باعث القای توقف چرخه سلولی و آپوپتوز در لوسمی میلوئیدی مزمن می‌گردد هماتولوژي Hematology مقاله اصیل پژوهشی Original Article <div dir="ltr" style="text-align: right;"><span style="font-size:10pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span b="" lang="AR-SA" mitra="" style="font-family:">زمینه و هدف:</span></b> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">بر اساس شواهد روزافزون، بیان غیر‌طبیعی </span></span><span dir="LTR" style="font-size:9.0pt">microRNA</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> ها با تومورزایی، گسترش تومور و عود در لوسمی‌ها، از جمله لوسمی میلوئید مزمن (</span></span><span dir="LTR" style="font-size:9.0pt">CML</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) مرتبط هستند. در این مطالعه اثر بیان بیش از حد </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> در سلول‌های </span></span><span dir="LTR" style="font-size:9.0pt">K562</span><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> بر توقف چرخه سلولی و القا آپوپتوز بررسی گردید.</span></span><span lang="AR-SA" style="font-size:11.0pt"><span style="font-family:"B Mitra""></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span b="" lang="AR-SA" mitra="" style="font-family:">روش تحقیق:</span></b><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> در این مطالعه بنیادی کاربردی، وکتور</span></span><span dir="LTR" style="font-size:9.0pt">pLenti-III-pre miR-155-GFP</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> برای افزایش بیان </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> در رده سلولی </span></span><span dir="LTR" style="font-size:9.0pt">K562</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> از طریق نوکلئوفکشن در مقایسه با وکتور </span></span><span dir="LTR" style="font-size:9.0pt">pLenti-III - Backbone-GFP</span><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> به‌عنوان گروه کنترل (</span></span><span dir="LTR" style="font-size:9.0pt">Backbone</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) مورداستفاده گرفت. از فلوسایتومتری برای تأیید بیان وکتور حاوی </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> در سلول‌های آسیب‌دیده استفاده شد. پس از ترانسفکشن، میزان بیان </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، </span></span><span dir="LTR" style="font-size:9.0pt">BAX</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، </span></span><span dir="LTR" style="font-size:9.0pt">BCL2</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، </span></span><span dir="LTR" style="font-size:9.0pt">CASP3</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> و </span></span><span dir="LTR" style="font-size:9.0pt">TP53</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> با روش </span></span><span dir="LTR" style="font-size:9.0pt">RT-qPCR</span><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> اندازه‌گیری شد. در بررسی چرخه سلولی از رنگ</span></span> <span lang="AR-SA" style="font-size:11.0pt"><span style="background:white"><span b="" mitra="" style="font-family:"><span style="color:#1f1f1f">پروپیدیوم </span></span></span></span><span lang="AR-SA" style="font-size:11.0pt"><span style="background:white"><span b="" mitra="" style="font-family:"><span style="color:black">یدید</span></span></span></span> <span dir="LTR" style="font-size:9.0pt"><span style="background:white"><span style="color:black">Propidium Iodide</span></span></span><span dir="LTR" style="font-size:12.0pt"> (</span><span dir="LTR" style="font-size:11.0pt">PI)</span><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> استفاده گردید و سلول</span></span><span lang="AR-SA" style="font-size:11.0pt">‌</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های رنگ</span></span><span lang="AR-SA" style="font-size:11.0pt">‌</span><span lang="AR-SA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">آمیزی شده توسط فلوسایتومتری قرائت شدند.</span></span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"><span style="color:black">اهمیت آماری نیز به عنوان مقدار </span></span></span><span dir="LTR" style="font-size:9.0pt"><span style="color:black">P</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"><span style="color:black"> کمتر از 05/0تعریف شد.</span></span></span><span lang="AR-SA" style="font-size:11.0pt"><span style="font-family:"B Mitra""></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span b="" lang="AR-SA" mitra="" style="font-family:">یافته‌ها:</span></b> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">پس از تأیید افزایش بیان </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، هنگامی که سلول‌های ترانسفکت‌شده با </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> با سلول‌های ترانسفکت‌شده با </span></span><a name="_Hlk154829281"><span dir="LTR" style="font-size:9.0pt">Backbone</span></a><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، شدند، توقف چرخه سلولی در گروه </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> مشاهده شد. در سلول‌های با افزایش بیان </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، تغییر در نسبت‌های </span></span><span dir="LTR" style="font-size:9.0pt">G0/S</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> و </span></span><span dir="LTR" style="font-size:9.0pt">G1/S</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> در مقایسه با </span></span><span dir="LTR" style="font-size:9.0pt">Backbone</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> به ترتیب به 5/7 و 5/4 نشان&shy;داده شد. 48 ساعت پس از ترانسفکشن، بیان ژن‌های </span></span><span dir="LTR" style="font-size:9.0pt">TP53</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، </span></span><span dir="LTR" style="font-size:9.0pt">BAX</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> و </span></span><span dir="LTR" style="font-size:9.0pt">CASP3</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> در گروه بیان کننده </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> افزایش و بیان </span></span><span dir="LTR" style="font-size:9.0pt">BCL2</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> کاهش یافت.</span></span><span lang="AR-SA" style="font-size:11.0pt"><span style="font-family:"B Mitra""></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span b="" lang="AR-SA" mitra="" style="font-family:">نتیجه‌گیری:</span></b> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">افزایش سطح </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> باعث توقف چرخه سلولی و افزایش سطح بیان ژن پرو آپوپتوز می‌شود که نشان می‌دهد </span></span><span dir="LTR" style="font-size:9.0pt">miR-155</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> رشد تومور را در </span></span><span dir="LTR" style="font-size:9.0pt">CML</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> مهار می‌کند.</span></span><span lang="AR-SA" style="font-size:11.0pt"><span style="font-family:"B Mitra""></span></span></span></span></span></span></div> <span style="font-size:10pt"><span new="" roman="" style="font-family:" times=""><b><span style="font-size:11.0pt">Background and Aims:</span></b><span style="font-size:11.0pt"> According to the increasing evidence, abnormal expression of microRNAs is associated with tumorigenesis, tumor spread, and relapse in leukemias, including chronic myeloid leukemia (CML). The present study aimed to investigate the effect of miR-155 overexpression in K562 cells on cell cycle arrest and apoptosis induction.</span><span dir="RTL" lang="AR-SA" style="font-size:11.0pt"><span style="font-family:"B Yagut""></span></span></span></span><br> <span style="font-size:10pt"><span new="" roman="" style="font-family:" times=""><b><span style="font-size:11.0pt">Materials and Methods:</span></b><span style="font-size:11.0pt"> The pLenti-III-pre miR-155-GFP vector was used to increase miR-155 expression in the K562 cell line through nucleofection compared to the pLenti-III-Backbone-GFP vector as a control group (Backbone). The flow cytometry method was employed to confirm the expression of the vector containing miR-155 in damaged cells. After transfection, the expression levels of miR-155, BAX, BCL2, CASP3, and TP53 were measured by the RT-qPCR method. The propidium iodide (PI) dye was used to study the cell cycle, and the stained cells were read by flow cytometry. The statistical significance was also defined as a P-value less than 0.05.</span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span new="" roman="" style="font-family:" times=""><b><span style="font-size:11.0pt">Results:</span></b><span style="font-size:11.0pt"> After confirming the increase in miR-155 expression when cells transfected with miR-155 were compared with cells transfected with Backbone, cell cycle arrest was observed in the miR-155 group. In the cells with increased expression of miR-155, the change in G0/S and G1/S ratios compared to Backbone was shown to be 7.5 and 4.5, respectively</span> <span style="font-size:11.0pt">(P<0.05). Moreover, 48 hours after transfection and in the group expressing miR-155, the expression of TP53, BAX, and CASP3 genes increased, and the expression of BCL2 decreased (P<0.05).</span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span new="" roman="" style="font-family:" times=""><b><span style="font-size:11.0pt">Conclusion:</span></b><span style="font-size:11.0pt"> The increase in the level of miR-155 caused cell cycle arrest and increased pro-apoptotic gene expression, which indicates that miR-155 inhibits tumor growth in CML.</span></span></span></span></span> آپوپتوز, BAX, BCL-2, CASP3, چرخه سلولی, لوسمی میلوژن مزمن, microRNA-155, TP53 Apoptosis, BAX, BCL-2, CASP3, Cell cycle, Chronic myelogenous leukemia, microRNA-155, TP53 79 90 http://journal.bums.ac.ir/browse.php?a_code=A-10-3567-1&slc_lang=fa&sid=1 Mahdi Edalati مهدی عدالتی edalati.m1982@gmail.com 8700319475328460048753 8700319475328460048753 No Department of Hematology and Blood Bank, Faculty of Paramedicine, Tehran University of Medical Sciences, Tehran, Iran گروه هماتولوژی و بانک خون، دانشکده پیراپزشکی، دانشگاه علوم پزشکی تهران، تهران، ایران Shaban Alizadeh شعبان علیزاده alizadeh1982@gmail.com 8700319475328460048754 8700319475328460048754 No Department of Hematology and Blood Bank, Faculty of Paramedicine, Tehran University of Medical Sciences, Tehran, Iran گروه هماتولوژی و بانک خون، دانشکده پیراپزشکی، دانشگاه علوم پزشکی تهران، تهران، ایران Asghar Abdoli اصغر عبدلی a_abdoli@pasteur.ac.ir 8700319475328460048755 8700319475328460048755 No Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran گروه هپاتیت و ایدز، انستیتو پاستور ایران، تهران، ایران Forouzan Karam فروزان کرم forouzankaram18@gmail.com 8700319475328460048756 8700319475328460048756 No Department of Hematology and Blood Bank, Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran گروه هماتولوژی و بانک خون، مرکز تحقیقات سلولی و مولکولی، دانشگاه علوم پزشکی بیرجند، بیرجند، ایران Aliakbar Shekarchi علی اکبر شکارچی shekarchiaa@gmail.com 8700319475328460048757 8700319475328460048757 No Department of Pathology, Tabriz University of Medical Sciences, Tabriz, Iran دپارتمان پاتولوژی، دانشگاه علوم پزشکی تبریز، تبریز، ایران Mahtab Sayadi مهتاب صیادی sayadi.mahtab@yahoo.com 8700319475328460048758 8700319475328460048758 Yes Department of Hematology and Blood Bank, Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran گروه هماتولوژی و بانک خون، مرکز تحقیقات سلولی و مولکولی، دانشگاه علوم پزشکی بیرجند، بیرجند، ایران