غربالگری ژرم لاین جهش Phe80Val ژن MLH1 در زنان مبتلا به سرطان تخمدان با تکنیک Tetra ARMS-PCR
محورهای موضوعی : نشانگر های زیستی پلاسما
1 - گروه ژنتیک، واحد تنکابن، دانشگاه آزاد اسلامی، تنکابن، ایران
کلید واژه: سرطان تخمدان, پروموتر, بیان ژن MLH1,
چکیده مقاله :
ﺳﺮﻃﺎن اﭘﯽﺗﻠﯿﺎل ﺗﺨﻤﺪان 90 درصد ﺑﺪﺧﯿﻤﯽ ﻫﺎي ﺗﺨﻤﺪان و 25 درصد ﺑﺪﺧﯿﻤﯽﻫﺎي ﺗﻨﺎﺳـﻠﯽ زﻧﺎن را ﺗﺸﮑﯿﻞ ﻣﯽدﻫﺪ و از آﻧﺠﺎ ﮐﻪ دﯾـرتشخیص داده می شود،ﺑﻪ ﻫﻤﯿﻦ دﻟﯿﻞ در ﺑﯿﻦ همه ی ﺑﺪﺧﯿﻤﯽ ﻫﺎي دﺳﺘﮕﺎه ﺗﻨﺎﺳﻠﯽ زﻧـﺎن از ﺑـﺎﻻﺗﺮﯾﻦ ﻣﻮارد ﻣﺮگ وﻣﯿﺮ ﺑﺮﺧﻮردار اﺳت. ﺗﻐﯿﯿﺮات microRNA که بیان ژن را کنترل میکند در این سرطان به چشم می خورد. تغییر در الگوی متیله شدن در ناحیه پروموتر ژن ها نیز از دیگر پیشامد های سرطان تخمدان است. در مطالعه حاضر نمونه های مورد آزمايش ازافراد بيمار و افراد سالم از آزمایشگاه جمع آوري گردید.جمعيت مورد مطالعه شامل 25فرد مبتلا به سرطان تخمدان و 25فرد سالم در سنین مختلف بوده است. نمونه خون افراد در ويال ٣سي سي حاوي EDTA جمع آوری و در دماي 20- درجه نگهداري شد.از روش کیت، برای استخراج DNA ژنومی از خون، استفاده شد. سپس ازجایگاه Phe80Val ژن MLH1 جهت بررسی پلی مورفیسم آن با روش ARMS-PCR Tetraاستفاده و همچنین یک جفت پرایمر جهت تایید نهایی ARMS-PCR Tetra طراحی گردید. از بین نمونه های مورد بررسی هیچکدام از نمونه ها موتاسیونی مشاهده نشد و دو عدد از نمونه ها تعیین توالی شدند و جایگاه نوکلئوتید بدون هیچ تغییری مشاهده شد.بطور کلی نتایج تحقیق حاضر نشان داد که هیچگونه ارتباطی بین موتاسیون ژن MLH1مربوط به Phe80Val و سرطان تخمدان در نمونه های حاضر وجود ندارد هرچند نیاز به بررسی روی نمونه های بیشتراز سایر مناطق ایران می باشد.
Epithelial ovarian cancer accounts for 90% of ovarian malignancies and 25% of female genital malignancies, and since it is diagnosed late, it has the highest mortality rate among all female genital malignancies. Changes in microRNA that controls gene expression are seen in this cancer. Changes in the methylation pattern in the promoter region of genes are also another occurrence of ovarian cancer. In the present study, the samples tested were collected from patients and healthy individuals from the laboratory. The study population included 25 individuals with ovarian cancer and 25 healthy individuals of different ages. Blood samples were collected in 3cc EDTA vials and stored at -20°C. A kit method was used to extract genomic DNA from blood. Then, rs63749990 of the MLH1 gene was used to examine its polymorphism using the ARMS-PCR Tetra method, and a pair of primers was designed for final confirmation by ARMS-PCR Tetra. No mutation was observed in any of the samples examined, and two of the samples were sent abroad for sequencing, and the nucleotide position was observed without any change. In general, the results of the present study showed that there is no association between the MLH1 gene mutation related to rs63749990 and ovarian cancer in the present samples, although there is a need to investigate more samples from other regions of Iran.
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