Characterization of primary normal and malignant breast cancer cell and their response to chemotherapy and immunostimulatory agents

Anna A Nushtaeva,Eugeniy I Savelyev,Alexey V Gerasimov,Olga A Koval,Grigory A Stepanov,Sergey V Sidorov,Dmitry V Semenov,Evgenia A Balahonova,Elena V Kuligina,Vladimir A Richter,Evgeny S Juravlev

doi:10.1186/s12885-018-4635-8

Abstract

BackgroundThe phenomenon of chemotherapy-resistant cancers has necessitated the development of new therapeutics as well as the identification of specific prognostic markers to predict the response to novel drugs. Primary cancer cells provide a model to study the multiplicity of tumourigenic transformation, to investigate alterations of the cellular response to various molecular stimuli, and to test therapeutics for cancer treatment.MethodsHere, we developed primary cultures of human breast tissue – normal cells (BN1), cancer cells (BC5), and cells from a chemotherapy-treated tumour (BrCCh1) to compare their response to conventional chemotherapeutics and to innate immunity stimulators with that of the immortalized breast cells MCF7, MDA-MB-231, and MCF10A. Expression of the progesterone receptor (PGR), oestrogen receptor (ER) α and β, human epidermal growth factor receptor (HER) 2 and 3 and aromatase CYP19, as well as expression of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) mRNA in human breast cells were characterized.ResultsWe revealed that BC5 carcinoma cells were PGRlow/ERbhigh/ERa−/Cyp19+, the BrCCh1 cells that originated from the recurrent tumour were PGR−/ERb+/ERa−/Cyp19+, and normal BN cells were PGR−/ERb+/ERa−/Cyp19high. The treatment of primary culture cells with antitumour therapeutics revealed that BrCCh1 cells were doxorubicine-resistant and sensitive to cisplatin. BC5 cells exhibited low sensitivity to tamoxifen and cisplatin. The innate immunity activators interferon-α and an artificial small nucleolar RNA analogue increased expression of IFIT3 at different levels in primary cells and in the immortalized breast cells MCF7, MDA-MB-231, and MCF10A. The relative level of activation of IFIT3 expression was inversely correlated with the baseline level of IFIT3 mRNA expression in breast cell lines.ConclusionOur data demonstrated that primary cancer cells are a useful model for the development of novel cancer treatments. Our findings suggest that expression of IFIT3 mRNA can be used as a prognostic marker of breast cancer cell sensitivity to immunostimulating therapeutics.

Highlights

The phenomenon of chemotherapy-resistant cancers has necessitated the development of new therapeutics as well as the identification of specific prognostic markers to predict the response to novel drugs
HER2 expression allows for the choice of anti-HER2 targeted therapy, whereas the triple negative breast cancer (TNBC) subtype is usually more aggressive, with the worst prognosis, as targeted or hormonal therapy is not available and patients are treated with standard chemotherapy or radiotherapy [6]
To assess whether the established cell cultures express Estrogen receptors (ER), progesterone receptor (PGR), and Cyp19, we analysed their specific mRNA compared with mRNA levels in the ER-positive human breast adenocarcinoma cell line MCF-7. mRNA signals from primary cells were defined as low/high if they were three times or more lower/higher than in MCF-7 cells, respectively

Summary

Introduction

The phenomenon of chemotherapy-resistant cancers has necessitated the development of new therapeutics as well as the identification of specific prognostic markers to predict the response to novel drugs. Cancer development is a multi-step process where various oncogenic mutations give rise to cancer cells with different genetic defects, which can differ even within an individual tumour. This diversity is a significant obstacle in cancer treatment, so cell lines established from human tumour samples can be a beneficial tool for screening relevant drugs. The significant characteristics of individual breast cancers are the expression of oestrogen, progesterone, and human epidermal growth factor receptor 2 (HER2/CD340), and the proposed type of treatment is based in part on these characteristics [3]. HER2 expression allows for the choice of anti-HER2 targeted therapy, whereas the triple negative breast cancer (TNBC) subtype is usually more aggressive, with the worst prognosis, as targeted or hormonal therapy is not available and patients are treated with standard chemotherapy or radiotherapy [6]

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Results

Discussion

Conclusion