Journal of Ilam University of Medical Sciences
مجله دانشگاه علوم پزشکی ایلام
J. Ilam Uni. Med. Sci.
Medical Sciences
http://sjimu.medilam.ac.ir
96
journal96
1563-4728
2588-3135
doi
fa
jalali
1403
2
1
gregorian
2024
5
1
32
2
online
1
fulltext
fa
بررسی آثار ژن کدکنندۀ buforin II بر بیان lncRNAهای PVT1، EGOT و LINC00312 در ردۀ سلولی سرطان کلیه
Investigating the effects of buforin II coding gene on the expression of lncRNAs PVT1, EGOT and LINC00312 in kidney cancer cell line
ژنتیک
Molecular Genetics
پژوهشي
Research
<div style="text-align: justify;"><span style="font-size:8pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">کارسینوم سلول کلیوی (</span></span>RCC<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">) شایع</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">ترین نوع سرطان کلیه در بزرگسالان است. هدف از مطالعۀ حاضر بررسی آثار ژن کدکنندۀ </span></span>buforin II<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> بر بیان </span></span>lncRNA<span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">های </span></span>PVT1<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">، </span></span>EGOT<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> و </span></span>LINC00312<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> در سلول</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">های سرطان کلیۀ ردۀ </span></span>ACHN<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> است.</span></span><span lang="AR-SA" style="font-size:9.0pt"><span style="font-family:"B Zar""></span></span></span></span></span></span><br>
<span style="font-size:8pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><b><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:.1pt">مواد و روش­ها:</span></span></span></span></b> <span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">پلاسمید نوترکیب </span></span>pcDNA3.1<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">(+) حاوی ژن </span></span>buforin II<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> و پلاسمید خالی </span></span>pcDNA3.1<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">(+) به روش شوک حرارتی وارد باکتری </span></span>E.coli<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> سویۀ </span></span>TOP10<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> شدند و سپس تخلیص گردیدند. پس از آن، هر دو پلاسمید به روش لیپوفکشن وارد سلول</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">های </span></span>ACHN<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> شدند و غربالگری توسط آنتی</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">بیوتیک نئومایسین انجام گرفت. درنهایت، واکنش </span></span>real time RT-PCR<span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> بهمنظور بررسی تغییر بیان </span></span>lncRNA<span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">های </span></span>EGOT<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">، </span></span>PVT1<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> و </span></span>LINC00312<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> انجام گردید.</span></span><span lang="AR-SA" style="font-size:9.0pt"><span style="font-family:"B Zar""></span></span></span></span></span></span><br>
<span style="font-size:8pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><b><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:.1pt">یافته­ های پژوهش: </span></span></span></span></b><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">پس از لیپوفکشن، سلول</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">های ترانسفکتشده در محیط کشت حاوی آنتیبیوتیک نئومایسین رشد کردند. واکنش </span></span>real time RT-PCR<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> نشان داد که بیان ژن </span></span>buforin II<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> در سلولهای سرطان کلیۀ </span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">ردۀ </span></span>ACHN<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> سبب افزایش بیان معنی</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">دار </span></span>lncRNA<span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">های </span></span>EGOT (P= 0.0033)<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> و </span></span>LINC00312 (P= 0.0272)<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> و کاهش بیان معنی</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">دار </span></span>PVT1 (P=0.0278)<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> شد.</span></span><b><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:.1pt"></span></span></span></span></b></span></span></span></span><br>
<span style="font-size:8pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><b><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.1pt">بحث و نتیجهگیری</span></span></span></span></b><span lang="AR-SA" style="font-size:9.0pt"><span style="font-family:"B Zar"">: </span></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">با توجه به اینکه حضور ژن </span></span>buforin II<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> باعث تغییر بیان معنی</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">دار </span></span>lncRNA<span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">های </span></span>EGOT<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">، </span></span>PVT1<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> و </span></span>LINC00312<span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar""> می</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">شود، احتمال می</span></span><span lang="FA" style="font-size:9.0pt"></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Zar"">رود که بتواند مسیرهای سلولی ازجمله آپتوزیز را فعال کند.</span></span></span></span></span></span><br>
<br>
</div>
<div style="text-align: justify;"><span style="font-size:9pt"><span style="font-family:"Times New Roman",serif"><b><span style="color:black"><span style="letter-spacing:-.2pt">Introduction:</span></span></b> Renal cell carcinoma (RCC) is the most common type of kidney cancer in adults. The aim of this study is to investigate the effects of the buforin II gene on the expression of lncRNAs PVT1, EGOT and LINC00312 in ACHN kidney cancer cells.</span></span><br>
<span style="font-size:9pt"><span style="font-family:"Times New Roman",serif"><b><span style="color:black"><span style="letter-spacing:-.2pt">Material & Methods: </span></span></b> Recombinant plasmid pcDNA3.1(+) containing buforin II gene and empty plasmid pcDNA3.1(+) were introduced into E. coli strain TOP10 by heat shock method and then purified. Then both mentioned plasmids were introduced into ACHN cells by lipofection method and screening was done by neomycin antibiotic. Finally, real time RT-PCR reaction was performed in order to investigate the expression change of EGOT, PVT1 and LINC00312 lncRNAs.<span style="font-size:12.0pt"><span style="color:#374151"></span></span></span></span><br>
<span style="font-size:9pt"><span style="font-family:"Times New Roman",serif"><b><span style="color:black"><span style="letter-spacing:-.2pt">Results:</span></span></b> After lipofection, the transfected cells grew in the culture medium containing neomycin antibiotic. The real time RT-PCR reaction showed that the expression of buforin II gene in ACHN kidney cancer cells caused a significant increase in the expression of lncRNAs EGOT (P=0.0033) and LINC00312 (P=0.0272) and a significant decrease in the expression of PVT1 (P=0.0278).</span></span><br>
<span style="font-size:9pt"><span style="font-family:"Times New Roman",serif"><b><span style="color:black"><span style="letter-spacing:-.2pt">Discussion & Conclusion:</span></span></b> Considering that the presence of buforin II gene causes a significant change in the expression of lncRNAs EGOT, PVT1 and LINC00312, it is possible that it can activate cell pathways including apoptosis.</span></span></div>
کارسینوم سلول کلیه, buforin II, ردۀ سلولی ACHN
Renal cell carcinoma, buforin II, ACHN cell line
32
43
http://sjimu.medilam.ac.ir/browse.php?a_code=A-10-7657-1&slc_lang=fa&sid=1
Maryam
Amiri-Farsani
مریم
امیری فارسانی
maryam.amirifarsani@yahoo.com
9600319475328460048048
9600319475328460048048
No
Dept of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
گروه زیستشناسی، دانشکدۀ علوم پایه، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران
Abbas
Doosti
عباس
دوستی
msafarpour84@yahoo.com
9600319475328460048047
9600319475328460048047
Yes
Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
مرکز تحقیقات بیوتکنولوژی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران