یافته های نوین در علوم زیستی

ارزیابی تغییرات بیان ژن های مسیر میتوکندریایی آپوپتوز و چرخه سلولی در سلول های AsPC-1 تیمار شده با فلورواوراسیل

نویسندگان

نگار خراسانی 1
جواد بهارآرا 2
خدیجه نژاد شاهرخ آبادی 1

1 گروه زیست شناسی، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران

2 گروه زیست شناسی و مرکز تحقیقات بیولوژی کاربردی تکوین جانوری، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران

چکیده
سرطان پانکراس یکی از کشند‌ه­ترین و تهاجمی‌­ترین سرطان­‌ها است؛ فلورواوراسیل باعث توقف در چرخه سلولی و القای آپوپتوز در سلول­های سرطانی می­شود. در مطالعه حاضر اثر فلورواوراسیل بر مراحل مختلف چرخه سلولی و بیان ژن­های درگیر در مسیر درونی آپوپتوز در رده سلولی AsPC-1 (سرطان پانکراس انسانی)، بررسی شده است. در این پژوهش تجربی-آزمایشگاهی برای بررسی اثر سمیت فلورواوراسیل بر روی تکثیر سلول­های AsPC-1 از آزمون MTT استفاده شد؛ نوع مرگ سلولی القا شده و تغییرات در چرخه سلولی به روش فلوسایتومتری بررسی شد؛ تغییرات در سطح بیان ژن­‌های (BAX، Bcl-2، APAF-1، Caspase-3، Caspase-9، p53 و p21) با تکنیک Real-time PCR ارزیابی شد. داده‌­های کمی حاصل در سطح معنی‌­داری (p< 0.05) تحلیل گردید. یافته­‌های حاصل از آزمون MTT نشان داد که فلورواوراسیل به­‌صورت وابسته به غلظت باعث کاهش تکثیر سلول­های AsPC-1 می­‌شود، نتایج آنالیز فلوسایتومتری نشان دهنده افزایش درصد سلول­‌های آپوپتوتیک در سلول­‌های تحت تیمار بود؛ فلورواوراسیل در سلول­های AsPC-1 سبب توقف چرخه سلولی در مرحله S و کاهش جمعیت سلولی در فاز G1 شد. بررسی نتایج Real-time PCR در سلول­‌های تیمار شده، نشان دهنده افزایش بیان ژن­های مسیر میتوکندریایی آپوپتوز و همچنین ژن­های موثر در تنظیم چرخه سلولی بود. فلورواوراسیل باعث کاهش تکثیر سلولی و القای آپوپتوز از طریق افزایش بیان ژن­های دخیل در مسیر درونی آپوپتوز در سلول­های AsPC-1 می­‌شود، همچنین فلورواوراسیل با تنظیم ژن­های (p53 و p21) سبب توقف چرخه سلولی در این سلول‌ها می­‌شود.


کلیدواژه‌ها

تکثیر سلولی
تکنیک Real-time PCR
سرطان پانکراس
فلوسایتومتری

عنوان مقاله English

Evaluation of genes expression changes in the mitochondrial pathway of apoptosis and cell cycle in AsPC-1 cells treated with Fluorouracil

نویسندگان English

Negar Khorasani 1
Javad Baharara 2
Khadijeh Nejad Shahrokhabadi 1
1 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
2 Department of Biology & Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
چکیده English

Pancreatic cancer is one of the most deadly and aggressive cancers; Fluorouracil induces apoptosis and cell cycle arrest in cancer cells. In the present study; the effect of Fluorouracil on different stages of the cell cycle and the expression of genes involved in the internal pathway of apoptosis in the AsPC-1 cell line (human pancreatic cancer) were investigated. In order to do so, MTT assay was used to evaluate the cytotoxic effect of Fluorouracil on AsPC-1 cell proliferation; The type of induced cell death and cell cycle changes were investigated by flow cytometry; changes in the expression level of genes (BAX, Bcl-2, APAF-1, Caspase-3, Caspase-9, p53, p21) were examined by Real-time PCR. Quantitative data were analyzed at the significant level of (p<0.05). The MTT assay results showed that Fluorouracil decreased AsPC-1 cell proliferation in a concentration-dependent manner. The results of flow cytometry analysis showed that increased percentage of apoptotic cells in the treated cells; Fluorouracil induces S phase cell cycle arrest in AsPC-1 cells and reduced distribution in the G1 phase. The Real-time PCR results in treated cells showed an increase in the expression of genes in the mitochondrial apoptotic pathway as well as genes effective in regulating the cell cycle. Fluorouracil reduces cell proliferation and induces apoptosis by increasing the expression of genes involved in the Intrinsic apoptotic pathway in AsPC-1 cells; Fluorouracil also caused cell cycle arrest in these cells by regulating the (p53, p21) genes.

کلیدواژه‌ها English

cell proliferation
flow cytometry
pancreatic cancer
real-time PCR
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  • تاریخ دریافت 18 خرداد 1405
  • تاریخ انتشار 18 خرداد 1405

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