Comparison of microrna145-5p expression level in breast cell lines

Mohammad Shayestehpour,1,* Shaghayegh yazdani,2 Talat mokhtari-azad,3

1. Autoimmune Diseases Research Center, Kashan University of Medical Sciences, Kashan, I.R. Iran
2. Department of Microbiology, Faculty of Advanced Science & Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
3. Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Introduction

Recent studies have reported that micrornas (mirnas) play critical roles in controlling cancer progression. mirna-145 is a tumor suppressive mirnas that down-regulates in cancer cells. there is not enough data on expression level of this pivotal mirna in some human cell lines to help researchers for selection of the best breast cancer cell line in them studies. therefore, the present study aimed to measure expression level of mirna145-5p in breast cell lines.

Methods

Hmepc, mcf-7, mcf-10a, mda-mb-231, mda-mb-453, mda-mb-468 and bt-20 cell lines were cultured in dulbecco modified eagle medium (dmem). small rnas were extracted from cells using high pure mirna isolation kit (roche, germany). one microgram of total rna was reverse transcribed into cdna using specific rt primers, and quantitative real-time pcr assay was performed. mirna expressions were normalized to small nuclear rna u6 expression.

Results

Mir145-5p was highly expressed in human mammary epithelial cells (hmepc) and mcf-10a as normal breast cells, but was significantly down-regulated in mcf-7, mda-mb-231, mda-mb-453, mda-mb-468, and bt-20 breast cancer cell lines. the concentration of mir-145-5p in hmepc was threefold higher than that of mcf-10a cells. the expression levels of mir-145-5p in mda-mb-453, mcf-7 and bt-20 were 0.012, 0.011 and 0.03, respectively, relative to hmepc cells. mir145-5p level in mda-mb-453 metastatic breast cancer cell line was 3 times lower than that in bt-20 non-metastatic cell line.

Conclusion

Hmepc has the highest level of mir145-5p that can be considered as a good normal breast cell line. mda-mb cells have low level of mir145-5p and can be the best option for mir-145 overexpression studies.

Keywords

Breast cancer, cell line, microrna