The Journal of Shahid Sadoughi University of Medical Sciences
مجله علمي پژوهشي دانشگاه علوم پزشكي شهید صدوقی يزد
JSSU
Medical Sciences
http://jssu.ssu.ac.ir
20
journal20
2228-5741
2228-5733
fa
jalali
1398
7
1
gregorian
2019
10
1
27
6
online
1
fulltext
fa
تغییر بیان ژن های آپوپتوزی BAK، FAS، BAX، TNF-a، BCL-2 و Survivin در سلول های آدنوکارسینومای پستان انسان (MCF-7) به وسیله انتروتوکسین B استافیلوکوکی
Change in the Expression of BAK، FAS، BAX، TNF-A، BCL-2 and Survivin Apoptosis Genes of the Human Breast Adenocarcinoma Cells (MCF-7) by Staphylococcal Enterotoxin B
زیست شناسی
Biology
پژوهشي
Original article
<strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">مقدمه:</span></span></strong> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">سرطان سینه یکی از شایع­ترین بدخیمی­ها در میان زنان می­باشد. مبتلایان به این عارضه در نتیجه استفاده از راهکارهای درمانی دارای پاسخ به درمان ضعیفی هستند و عود مجدد در میان آنها به فراوانی دیده میشود. در این مطالعه اثر انتروتوکسین </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">B</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> استافیلوکوکی بر بیان ژن­های </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BAK</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">FAS</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BAX</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF-a</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BCL-2</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> و </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">Survivin</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> در سلول­های آدنوکارسینومای پستان انسان (</span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">MCF-7</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">) مورد بررسی قرار گرفت.</span></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> <span style="color:black;">انتروتوکسین </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">B</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> استافیلوکوکی</span></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> عضو توانمندی از خانواده سموم <em>استافیلوکوکوس اورئوس</em> می­باشد که به عنوان یک عامل ضد سرطان با توانایی انهدام سلول­های سرطانی شناخته می­شود.</span></span><br>
<strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">روش بررسی:</span></span></strong> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">این مطالعه به صورت تجربی در مرکز تحقیقات بیوتکنولوژی دانشگاه آزاد اسلامی</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">واحد شهرکرد انجام شد. سلول­های رده </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;"> MCF-7</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">با استفاده از معرف لیپوفکتامین 2000، بهوسیله پلاسمیدهای </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">pcDNA3.1(+)-<em>seb</em></span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> (نوترکیب) و </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">pcDNA3.1(+)</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> (غیرنوترکیب) ترانسفورم شدند و با کشت در محیط انتخابی </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">RPMI-1640</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> محتوی</span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">600 μg/mL </span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> آنتیبیوتیک </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">G418</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> انتخاب گردیدند. سپس میزان بیان ژن­های </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BAK</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">FAS</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BAX</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF-a</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BCL-2</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">Survivin</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> در سلول­های ترانسفکت شده از طریق آنالیز </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">Real Time-PCR</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> مورد بررسی قرار گرفت. آنالیز آماری با استفاده از آزمون تی- استیودنت از طریق نرمافزار </span></span></span><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">SPSS Inc., Chicago, IL; Version </span></span></span><span dir="LTR"><span style="font-family:times new roman,serif;">16</span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">و همچنین برنامه </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">Excel</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> مورد بررسی قرار گرفت.</span></span></span><br>
<strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">نتایج:</span></span></strong> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">نتایج حاصل از این تحقیق حاکی از آن است که، انتروتوکسین </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">B</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> استافیلوکوکی بهطور قابلتوجهی بیان ژن­های آپوپتوزی را در سلول­های رده </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">MCF-7</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> تغییر می­دهد. در حالیکه افزایش بیان معنی­داری برای ژن­های </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BAK</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">FAS</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BAX</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF-a</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، مشاهده گردید، بیان ژن­های </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BCL-2</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span></span><em><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">Survivin</span></span></span></span></em><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> بهطور معنی­داری کاهش را در مقایسه با گروه کنترل نشان داد.</span></span></span> <span style="font-family:b nazanin;"><span style="font-size:12.0pt;">(032/</span></span><em><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">P=</span></span></span></em><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">)</span></span><br>
<strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">نتیجهگیری:</span></span></strong> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">انتروتوکسین </span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">B</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> استافیلوکوکی دارای</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">اثر</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">مهاری</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">بر</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">رشد،</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">تکثیر</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">و</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">تهاجم</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">سلول­های</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">آدنوکارسینومای پستان از</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">طریق</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">تغییر بیان</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">ژن­های</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">دخیل</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">در</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">مسیر</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">آپوپتوز</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">می­باشد</span></span></span><span dir="LTR"><span style="color:black;"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">.</span></span></span></span><span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> لذا به</span></span></span> <span style="color:black;"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">نظر می­رسد زمینه تحقیقاتی مناسبی برای بهره برداری از این سم در کنترل و درمان آدنوکارسینومای پستان انسان وجود داشته باشد</span></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">.</span></span><br>
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<p class="MsoNormal" style="line-height:150%"><strong>Introduction:</strong> Breast cancer is one of the most prevalent malignancies among women. Patients whose suffering from this condition, as a result of the use of conventional therapies often have a poor response to treatment and the relapse among them is frequent. In this study, the effects of staphylococcal enterotoxin type B on <em>BAK</em><span dir="RTL">، </span><em>FAS</em><span dir="RTL">، </span><em>BAX</em><span dir="RTL">، </span><em>TNF-a</em><span dir="RTL">، </span><em>BCL-2</em><span dir="RTL"> و </span><em>Survivin</em> genes expression in human breast adenocarcinoma cells (MCF-7) was examined. Staphylococcal enterotoxin B is a powerful member of the Staphylococcus aureus toxins family, which is known as an anticancer agent with potential for killing cancer cells.<br>
<strong>Methods:</strong> The experimental study was carried out at the Biotechnology Research Center of Islamic Azad University, Shahrekord Branch. By using Lipofectamine 2000 reagent, MCF-7 cells transfected with the pcDNA3.1(+)-seb (recombinant) and pcDNA3.1(+) (non-recombinant) plasmids and were selected by culturing in a selective medium of RPMI- 1640 containing 600 μg / mL antibiotic G418. Then, the expression of <em>BAK, FAS, BAX, TNF-a, BCL-2,</em> and <em>Survivin</em> genes in transfected cells were analyzed by real time PCR. Student's t-test, SPSS Inc., Chicago, IL; Version 16 and also Excel program for statistical analysis were used.<br>
<strong>Results:</strong> The results of this study indicated that staphylococcal enterotoxin type B (SEB) remarkably changes the expression of apoptotic related genes in MCF-7 cell line. It was observed a significant increase in the expression of <em>BAK, FAS, BAX, </em>and<em> TNF-a</em> genes, the expression of <em>BCL-2 </em>and<em> Survivin</em> genes significantly decreased compared to the control group (<em>P</em>=0/032).<br>
<strong>Conclusion:</strong> Staphylococcal enterotoxin type B has an inhibitory effect on the growth, proliferation and invasion of breast adenocarcinoma cells through altering the expression of the genes involved in the apoptosis process. Therefore, it seems that there is a good research field for the use of this toxin in the control and treatment of human breast adenocarcinoma.<br>
</p>
سرطان پستان, انتروتوکسین B, آپوپتوز, MCF-7, BAK, FAS, BAX, TNF-a, BCL-2, Survivin
Breast cancer, Enterotoxin B, Apoptosis, MCF-7, BAK, FAS, BAX, TNF-a, BCL-2, Survivin
1677
1685
http://jssu.ssu.ac.ir/browse.php?a_code=A-10-3994-1&slc_lang=fa&sid=1
Sara
Afzali
سارا
افضلی
sara.afzali.dinani@gmail.com