Characterization of microRNA-125b expression in MCF7 breast cancer cells by ATR-FTIR spectroscopy

Date

2010-01-01

Author

OZEK, Nihal Simsek
TUNA, Serkan
Erson Bensan, Ayşe Elif
Severcan, FERİDE

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

200
views

0
downloads

MicroRNAs (miRNAs), are similar to 22 nucleotides long, non-coding RNAs that control gene expression post-transcriptionally by binding to their target mRNA's 3'UTRs (untranslated regions). Due to their roles in various important regulatory processes and pathways, miRNAs have been implicated in disease mechanisms such as tumorigenesis when their expression is deregulated. To date, a significant number of miRNAs and their target messenger RNAs (mRNAs) have been identified and verified. It is generally accepted that miRNAs can potentially bind to many mRNAs, which brings the requirement of validation of these interactions. While understanding that such individual interactions is crucial to delineate the role of a specific miRNA, we took a holistic approach and analyzed global changes in the cell due to expression of a miRNA in a model cell line system. Our model consisted of MCF7 cells stably transfected with miR-125b (MCF7-125b) and empty vector (MCF7-EV). MiR-125b is one of the known down-regulated miRNAs in breast cancers. In this study we examined the global structural changes in MCF7 cells lacking and expressing miR-125b by Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) Spectroscopy and investigated the dynamic changes by more sensitive spin-labelling Electron Spin Resonance (ESR) spectroscopy. Our results revealed less RNA, protein, lipid, and glycogen content in MCF7-125b compared to MCF7-EV cells. Membrane fluidity and proliferation rate were shown to be lower in MCF7-125b cells. Based on these changes, MCF7-125b and MCF7-EV cells were discriminated successfully by cluster analysis. Here, we provide a novel means to understand the global effects of miRNAs in cells. Potential applications of this approach are not only limited to research purposes. Such a strategy is also promising to pioneer the development of future diagnostic tools for deregulated miRNA expression in patient samples.

Subject Keywords

K562 cells, Low-densıty, Animal-model, Human tissues, Model membranes; , Membrane fluidity, Ir spectroscopy; , Unsaturated fatty-acids, Induced lipid-peroxidation;, Transform-infrared-spectroscopy

URI

https://hdl.handle.net/11511/32711

Journal

ANALYST

DOI

https://doi.org/10.1039/c0an00543f

Collections

Department of Biology, Article

Suggestions

OpenMETU
Core

Characterization of molecular-level changes due to MicroRNA-125B expression in breast cancer cells by spectroscopic and chemometric analysis techniques Şimşek Özek, Nihal; Severcan, Feride; Erson Bensan, Ayşe Elif; Department of Biology (2015) MicroRNAs (miRNAs), are 22 nucleotides long, non-coding RNAs that control gene expression post-transcriptionally by binding to their target mRNA’s 3’ UTRs (untranslated regions). Due to their crucial roles in various important regulatory processes and pathways, miRNAs have been implicated in disease mechanisms such as tumorigenesis, metastasis and drug resistance when their expression is deregulated. Moreover, the regulatory roles of these micromanagers have been demonstrated in the metabolism of cancer cel...
Analysis of motifs in microRNA-transcription factor gene regulatory networks Sürün, Bilge; Acar, Aybar Can; Purutçuoğlu Gazi, Vilda; Department of Bioinformatics (2014) MicroRNAs are small non-coding RNA molecules which contain 21-25 nucleotides, and function in post transcriptional regulation by inhibiting the translation of mRNA targets. miRNAs typically affect gene regulation by forming composite feed forward circuits (cFFCs) which also comprise a transcription factor (TF) and a target gene. By analyzing these cFFCs, the contribution of miRNAs in altering TF networks can be revealed. These contributions could either be the de-escalation of the target gene repertoire or ...
Investigation of human promoter CpG content and methylation profiles at different conservation levels Demiralay, Burak; Aydın Son, Yeşim; Department of Bioinformatics (2012) Methylation of CpG islands located at the promoter regions is a mechanism which controls gene silencing and expression. Hyper or hypo methylation of these sites on promoter sequences have been associated with many diseases, like cancer. Even though promoter CpG islands and their methylation profiles are important regulators of gene expression, the exact mechanism of gene silencing through methylation is not known. Here, we have investigated the status of promoter CpG methylation under various evolutionary p...
Phenotypical characterization of microRNA-106B overexpression in MCF10A breast cell line Saygılı, Cansaran; Erson Bensan, Ayşe Elif; Department of Biology (2013) MicroRNAs are small non-coding RNAs which regulate gene expression by binding to 3’UTR of their target mRNAs. Deregulated expression of microRNAs is detected in many pathologies including different types of cancers. miR-106b, is a member of miR-106b-25 cluster and overexpressed in many cancers including breast cancer. Based on miR-106b overexpression, we hypothesized that miR-106b may be an oncogene candidate. To explore miR-106b related phenotypes, we used an already miR-106b transfected model cell line sy...
Design of a sequence based miRNA clustering method; analysis of fungal miRNAs and host organism target genes Narcı, Kübra; Akkaya, Mahinur S.; Oğul, Hasan; Department of Bioinformatics (2014) MicroRNAs are small non-coding RNA molecules which contain 21-25 nucleotides, and function in post transcriptional regulation by inhibiting the translation of mRNA targets. miRNAs typically affect gene regulation by forming composite feed forward circuits (cFFCs) which also comprise a transcription factor (TF) and a target gene. By analyzing these cFFCs, the contribution of miRNAs in altering TF networks can be revealed. These contributions could either be the de-escalation of the target gene repertoire or ...

Citation Formats

N. S. OZEK, S. TUNA, A. E. Erson Bensan, and F. Severcan, “Characterization of microRNA-125b expression in MCF7 breast cancer cells by ATR-FTIR spectroscopy,” ANALYST, pp. 3094–3102, 2010, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/32711.