Tehran University Medical Journal
مجله دانشکده پزشکی، دانشگاه علوم پزشکی تهران
Tehran Univ Med J
Medical Sciences
http://tumj.tums.ac.ir
1
admin
1683-1764
1735-7322
10.18869/acadpub.tumj
000
000
000
fa
jalali
1397
7
1
gregorian
2018
10
1
76
7
online
1
fulltext
other
بررسی ارتباط بین تغییرات بیان LncRNA THRIL کنترلکننده مسیر TNF-alpha در رده سلولی گلیوبلاستومای تحت تیمار با تموزولامید
Relationship between LncRNA THRIL expression controlling TNF-alpha pathway in glioblastoma cell line under temozolomide treatment
مقاله اصیل
Original Article
<div style="text-align: justify;"><span style="font-family:yekanyw;"><strong><span style="font-size:10.0pt;">زمینه و هدف:</span></strong> <span style="color:black;"><span style="font-size:10.0pt;">گلیوما</span></span><span style="color:black;"><span style="font-size:10.0pt;">،</span></span><span style="color:black;"><span style="font-size:10.0pt;"> شایعترین و کشندهترین تومور بدخیم مغزی است. </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">LncRNA THRIL</span></span></span><span style="color:black;"><span style="font-size:10.0pt;">، از </span></span><span style="color:black;"><span style="font-size:10.0pt;">نوع آنتی سنس و </span></span><span style="color:black;"><span style="font-size:10.0pt;">واسطه مهم مسیر سیگنالی </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">NF-KB</span></span></span><span style="color:black;"><span style="font-size:10.0pt;">، که در بافتها از جمله سیستم عصبی </span></span><span style="color:black;"><span style="font-size:10.0pt;">مرکزی</span></span><span style="color:black;"><span style="font-size:10.0pt;"> بیان میگردد. </span></span><span style="color:black;"><span style="font-size:10.0pt;">بخش عمده فرآورده بیان ژن را </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">LncRNA</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> تشکیل میدهد. </span></span><span style="color:black;"><span style="font-size:10.0pt;">هدف مطالعه</span></span><span style="color:black;"><span style="font-size:10.0pt;">،</span></span><span style="color:black;"><span style="font-size:10.0pt;"> بررسی تغییرات میزان بیان </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">LncRNA THRIL</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> در آدنوکارسینومای گلیوبلاستوما</span></span><span style="color:black;"><span style="font-size:10.0pt;">،</span></span><span style="color:black;"><span style="font-size:10.0pt;"> رده سلولی </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">T98G</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> تحت تیمار با داروی شیمیدرمانی تموزولامید بود.</span></span><span style="font-size:10.0pt;"></span><br>
<strong><span style="font-size:10.0pt;">روش بررسی: </span></strong><span style="color:black;"><span style="font-size:10.0pt;">مطالعه کنونی از نوع مورد-شاهدی</span></span><span style="color:black;"><span style="font-size:10.0pt;"> از فروردین تا شهریور </span></span><span style="color:black;"><span style="font-size:10.0pt;">۱۳۹۶</span></span><span style="color:black;"><span style="font-size:10.0pt;"> در مرکز علوم تحقیقات دانشگاه آزاد اسلامی زنجان انجام شده است. رده سلولی </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">T98G</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> در غلظتهای متفاوت داروی تموزولامید (</span></span><span style="color:black;"><span style="font-size:10.0pt;">۲۵</span></span><span style="color:black;"><span style="font-size:10.0pt;">، </span></span><span style="color:black;"><span style="font-size:10.0pt;">۵۰</span></span><span style="color:black;"><span style="font-size:10.0pt;"> و </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">μM</span></span></span> <span style="color:black;"><span style="font-size:10.0pt;">۱۰۰</span></span><span style="color:black;"><span style="font-size:10.0pt;">) و در زمانهای مختلف (</span></span><span style="color:black;"><span style="font-size:10.0pt;">۲۴</span></span><span style="color:black;"><span style="font-size:10.0pt;">،</span></span> <span style="color:black;"><span style="font-size:10.0pt;">۴۸</span></span><span style="color:black;"><span style="font-size:10.0pt;"> و </span></span><span style="color:black;"><span style="font-size:10.0pt;">۷۲</span></span><span style="color:black;"><span style="font-size:10.0pt;"> ساعت) تیمار شده</span></span><span style="font-size:10.0pt;">، بهترتیب</span><span style="color:black;"><span style="font-size:10.0pt;"> استخراج </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">RNA</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> و سنتز </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">cDNA</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> انجام شد</span></span><span style="color:black;"><span style="font-size:10.0pt;">. سپس</span></span><span style="color:black;"><span style="font-size:10.0pt;"> بیان ژن </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">LncRNA THRIL</span></span></span> <span style="color:black;"><span style="font-size:10.0pt;">توسط </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">Real-time PCR</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> مورد ارزیابی قرار گرفت و</span></span> <span style="color:black;"><span style="font-size:10.0pt;">نتایج توسط روش کمیتسنجی نسبی و روش لیواک </span></span><span dir="LTR"><span style="font-size:8.0pt;">(Livak method)</span></span><span style="color:black;"><span style="font-size:10.0pt;"> آنالیز گردید.</span></span><span style="font-size:10.0pt;"></span><br>
<strong><span style="font-size:10.0pt;">یافتهها:</span></strong> <span style="color:black;"><span style="font-size:10.0pt;">بیان ژن </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">THRIL</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> در زمان ۲۴ ساعت</span></span><span style="color:black;"><span style="font-size:10.0pt;"> در</span></span><span style="color:black;"><span style="font-size:10.0pt;"> غلظتهای ۲۵ و </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">μM</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> ۱۰۰ (۰/۰۰۱</span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">P<</span></span></span><span style="color:black;"><span style="font-size:10.0pt;">) کاهش معنادار مشاهده گردید. زمان ۴۸ ساعت</span></span><span style="color:black;"><span style="font-size:10.0pt;"> در</span></span><span style="color:black;"><span style="font-size:10.0pt;"> غلظت </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">μM</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> ۵۰ (۰۰۱/۰</span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">P<</span></span></span><span style="color:black;"><span style="font-size:10.0pt;">) به استثنای غلظت ۲۵ و </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">μM</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> ۱۰۰ (۰/۰۰۱</span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">P<</span></span></span><span style="color:black;"><span style="font-size:10.0pt;">) افزایش معنادار مشاهده گردید. زمان ۷۲ ساعت</span></span><span style="color:black;"><span style="font-size:10.0pt;"> در </span></span><span style="color:black;"><span style="font-size:10.0pt;">غلظت </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">μM</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> ۵۰ (۰/۰۰۱</span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">P<</span></span></span><span style="color:black;"><span style="font-size:10.0pt;">) افزایش معنادار داشته است و در مقابل</span></span><span style="color:black;"><span style="font-size:10.0pt;">،</span></span><span style="color:black;"><span style="font-size:10.0pt;"> غلظتهای ۲۵ و </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">μM</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> ۱۰۰ (۰/۰۰۱</span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">P<</span></span></span><span style="color:black;"><span style="font-size:10.0pt;">) نشاندهنده کاهش بیان ژن </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">THRIL</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> بودهاند.</span></span><span style="font-size:10.0pt;"></span><br>
<strong><span style="font-size:10.0pt;">نتیجهگیری: </span></strong><span style="color:black;"><span style="font-size:10.0pt;">در نتیجه</span></span><span style="color:black;"><span style="font-size:10.0pt;">،</span></span><span style="color:black;"><span style="font-size:10.0pt;"> تغییرات بیان ژن </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">THRIL</span></span></span><span style="color:black;"><span style="font-size:10.0pt;"> پس از تیمار</span></span><span style="color:black;"><span style="font-size:10.0pt;">،</span></span><span style="color:black;"><span style="font-size:10.0pt;"> به زمان و غلظت دارو وابسته است و</span></span><span style="color:black;"><span style="font-size:10.0pt;"> در غلظت </span></span><span dir="LTR"><span style="color:black;"><span style="font-size:8.0pt;">μM</span></span></span> <span style="color:black;"><span style="font-size:10.0pt;">۵۰</span></span><span style="color:black;"><span style="font-size:10.0pt;"> و زمان </span></span><span style="color:black;"><span style="font-size:10.0pt;">۴۸</span></span><span style="color:black;"><span style="font-size:10.0pt;"> ساعت بالاترین تاثیر بر سلول با توجه به بیان </span></span><span style="color:black;"><span style="font-size:10.0pt;">ژن</span></span><span style="color:black;"><span style="font-size:10.0pt;"> داشته است</span></span><span style="color:black;"><span style="font-size:10.0pt;">.</span></span></span><span style="font-family:b lotus;"><span style="font-size:10.0pt;"></span></span></div>
<div style="text-align: justify;"><strong>Background</strong>: Glioma is one of the most common and deadliest primary malignant tumors in the brain. A large part of the gene expression products are non-coding protein RNA. LncRNA THRIL gene is an antisense LncRNA and one of the most important mediators of the NF-KB signaling pathway, that express in many tissues of the body, including the central nerve system (CNS). The aim of the present study was to investigate the alternation in the expression of LncRNA THRIL gene in cells of the adenocarcinoma glioblastoma T98G cell line, under treatment with temozolomide chemotherapy drug.<br>
<strong>Methods</strong>: This case-control study was conducted in Research Center of Islamic Azad University of Zanjan, Iran, from April to September 2017. Cells of T98G cell line was treated with various doses of temozolomide chemotherapy drug (25-50-100 μM) and at different times (72-48-48 hours), Respectively RNA extraction and cDNA synthesis were performed. Then LncRNA THRIL gene expression was studied by real-time PCR method and the results were analyzed by relative quantitative and livak methods.<br>
<strong>Results</strong>: The THRIL gene expression in 24 hours’ time with 25 and 100 μM doses (P< 0.001) had significant decreased expression and 50 μM dose had non-significant increase. It had significant increase in 48 hours time with 50 μM dose (P< 0.001), except 25 μM (P< 0.001) and 100 μM (P< 0.001) doses had significant decreased expression. It had significant increase during 72 hours time with 50 μM dose (P< 0.001), in contrast 25 μM (P> 0.001) and 100 μM (P< 0.001) concentrations of temozolomide chemotherapy drug demonstrated decreased expression of the LncRNA THRIL gene (P< 0.001).<br>
<strong>Conclusion</strong>: As a result, the THRIL gene expression alterations after cancer cells treatment by temozolomide chemotherapy drug depends on the time and dose of the drug and the 48 hours treatment time with 50 μM dose (P< 0.001) had the highest effect on cancer cells of the T98G cell line, due to the expression of the THRIL gene.</div>
LncRNA THRIL, واکنش زنجیرهای پلیمراز, TNF-, T98G
LncRNA THRIL, polymerase chain reaction, TNF-α, T98G
469
476
http://tumj.tums.ac.ir/browse.php?a_code=A-10-2748-2&slc_lang=other&sid=1
Leila
Hassani
لیلا
حسنی