بررسی بیان ژن DNase فعال شونده توسط کاسپاز (CAD) در رده ی سلولی سرطان پستان MCF-7 تیمار شده با نانو ذرات نقره زیستی

نویسندگان
گروه زیست شناسی، واحد تهران مرکزی، دانشگاه آزاد اسلامی، تهران، ایران
چکیده
مقدمه

سرطان پستان رایج‌ترین سرطان در میان زنان و دومین علت رایج مرگ ناشی از سرطان‌ها می‌باشد. به علت مقاومت اکتسابی سلول‌های سرطانی به درمان‌های رایج و عود مجدد بیماری پس از درمان، استراتژی‌های درمانی نوین براساس راه اندازی مجدد آپوپتوز مطرح شدند. هدف از انجام مطالعه حاضر، بررسی اثرات سمیت نانوذرات نقره زیستی و ارزیابی بیان ژنCAD (DNase فعال شونده توسط کاسپاز) بر روی سلول‌های سرطانی پستان (MCF-7) می‌باشد.

مواد و روش­ها

سنتز نانوذرات نقره با استفاده از عصاره اتانولی برگ گیاه درمنه آرتمیزیا کوتنسیس مطابق مطالعات گذشته انجام گرفت. اندازه و ریخت‌شناسی نانوذرات با کمک میکروسکوپ الکترونی عبوری (TEM) بررسی شد. رده سلولی MCF-7 در غلظت‌های مختلف 25/6، 5/12 ، 25 ، 50 ،100 و 200 µg/ml تیمار 24 ساعته نانوذرات نقره سنتز شده انجام گرفت و با کمک روش MTT ارزیابی و میزان 50 درصد کشندگی IC50 محاسبه شد. میزان بیان ژن CAD با روش PCR time Real ارزیابی شد.

یافته­ها

اندازه متوسط نانوذرات با کمک میکروسکوپ TEM حدود 86/4 ± 13/9 نانومتر بدست آمد. نانو ذرات نقره دارای اثرات سمیت به صورت وابسته به دوز بر روی رده سلولیMCF-7 بود. میزان IC50 نانوذرات نقره µg/ml 49 محاسبه شد. میزان بیان ژنCAD در نمونه تیمار نسبت به کنترل به میزان 77/0 برابر کاهش( 05/0P <) نشان داد.

نتیجه­گیری

ارزیابی نتایج این تحقیق نشان داد که نانوذرات نقره اثراث سمیت قابل قبولی بر روی رده سرطانی پستان دارد و نیاز به انجام آزمایشات بیشتر در مدل‌های حیوانی می‌باشد.
کلیدواژه‌ها

عنوان مقاله English

Assessment of caspase activated DNase (CAD) gene expression on MCF-7 breast cancer cell line treated by biosynthesized silver-nano particles

نویسندگان English

Fatemeh Moharrami
Seyed Ataollah Sadat Shandiz
Aras Rafiee
Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran
چکیده English

Introduction

Breast Cancer is the most prevalent type of cancer among women and is also the second major cause of cancer. Due to the acquired resistance of cancer cells against conventional therapies and recurrence of the sickness after treatment, new therapeutic strategies based on triggering apoptosis have been proposed. The purpose of the present study is to introduce a cytotoxic effect of biological silver nanoparticles toward breast cancer MCF-7 cells and evaluation of the caspase-activated DNase (CAD) gene expression at mRNA level.

Materials and Methods

Silver nanoparticles were synthesized according to the recent biosynthesize method. Afterward, the Silver nanoparticles were inspected using transmission electron microscopy (TEM) analysis. Different concentrations of 6.25, 12.5, 25, 50, 100, and 200 µg/ml of Nanosilver particles were treated on MCF-7 cells for 24 hours and the results were evaluated using the MTT assay and IC50 were measured. The level expression of CAD gene was evaluated using real-time PCR.

Results

Silver nanoparticles were mostly spherical, having an average size of 9.13±4.86 nm. Silver nanoparticles demonstrated a dose-dependent toxic effect toward MCF-7 cells. The IC50 value of silver nanoparticles was measured 49μg/mL. In addition, the expression of CAD decreased by 0.77 fold, compared to the control group.

Conclusion

The results of the present study demonstrated that the silver nanoparticles can induce cytotoxicity against breast cells and it is necessary to perform more tests against animal models.

کلیدواژه‌ها English

Apoptosis
Breast Cancer
CAD
Silver nano particles
Real time PCR
Cytotoxicity
Introduction
Breast Cancer is the most prevalent type of cancer among women and is also the second major cause of cancer. Due to the acquired resistance of cancer cells against conventional therapies and recurrence of the sickness after treatment, new therapeutic strategies based on triggering apoptosis have been proposed. The purpose of the present study is to introduce a cytotoxic effect of biological silver nanoparticles toward breast cancer MCF-7 cells and evaluation of the caspase-activated DNase (CAD) gene expression at mRNA level.
Materials and Methods
Silver nanoparticles were synthesized according to the recent biosynthesize method. Afterward, the Silver nanoparticles were inspected using transmission electron microscopy (TEM) analysis. Different concentrations of 6.25, 12.5, 25, 50, 100, and 200 µg/ml of Nanosilver particles were treated on MCF-7 cells for 24 hours and the results were evaluated using the MTT assay and IC50 were measured. The level expression of CAD gene was evaluated using real-time PCR.
Results
Silver nanoparticles were mostly spherical, having an average size of 9.13±4.86 nm. Silver nanoparticles demonstrated a dose-dependent toxic effect toward MCF-7 cells. The IC50 value of silver nanoparticles was measured 49μg/mL. In addition, the expression of CAD decreased by 0.77 fold, compared to the control group.
Conclusion
The results of the present study demonstrated that the silver nanoparticles can induce cytotoxicity against breast cells and it is necessary to perform more tests against animal models.