Anticancer Drug Screening Based On Label-Free Cytochrome C Assay By Fluorescent DNA Probe

Publish Year: 1395
نوع سند: مقاله کنفرانسی
زبان: English
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NASTARANCANSER02_155

تاریخ نمایه سازی: 22 دی 1396

Abstract:

Anticancer drugs mainly kill tumor cells via inducing intrinsic apoptosis. This process primarily isinitiated by release of Cytochrome c (Cyt c) from mitochondrial into the cytosol and thereuponapoptosome formation. Therefore, the release of Cyt c is perceived as a robust signal for detection ofearly stage apoptosis and evaluation of anti-cancer agents. Cyt c is typically recognized by Westernblot and ELISA, which are semi-quantitative, time-consuming and tedious methods. Fluorescentmetal nanoclusters (NCs), have received highly attention in the past few years owing to their greatpromise for highly sensitive detections. Here, we have designed and fabricated a novel biosensorbased on aptamer-stabilized nanoclusters for rapid and inexpensive evaluation of anticancer drugs.Oligonucleotide sequences were chemically synthesized and used for growth of fluorescentDNA/AgNCs Quantitative analysis of release of Cyt c from mitochondria was performed by additionof drug treated cells to DNA/AgNCs After for 20 min incubation, the fluorescence intensities weremeasured by Spectrofluorometer under excitation wavelength of 560 nm. The aptamer-stabilizedAg NCs could be used for qualitative and quantitative determination of label-free Cyt c with thelinear range from 0 to 1 μM and a detection limit of 15 nM. In this study, we have developed anaptamer-based fluorescent probe screen anti-cancer drugs. The resulting data suggest that thedesigned sensing platform can be used for monitoring of Cyt c in the apoptotic cells after theexposure to an anti-cancer drug (e.g. doxorubicin).

Authors

Fatemeh Molaabasi

Department Of Chemistry, Tarbiat Modares University, Tehran, Iran

Mojtaba Shamsipur

Department Of Chemistry, Razi University, Kermanshah, Iran

Saman Hosseinkhani

Department Of Biochemistry, Tarbiat Modares University, Tehran, Iran