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Curcumin prevented human autocrine growth hormone (GH) signaling mediated NF-κB activation and miR-183-96-182 cluster stimulated epithelial mesenchymal transition in T47D breast cancer cells

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Abstract

Autocrine growth hormone (GH) signaling is a promoting factor for breast cancer via triggering abnormal cell growth, proliferation, and metastasis, drug resistance. Curcumin (diferuloylmethane), a polyphenol derived from turmeric (Curcuma longa), has anti-proliferative, anti-carcinogenic, anti-hormonal effect via acting on PI3K/Akt, NF-κB and JAK/STAT signaling. Forced GH expression induced epithelial mesenchymal transition (EMT) through stimulation of miR-182-96-183 cluster expression in breast cancer cells. This study aimed to investigate the role of NF-κB signaling and miR-182-96-183 cluster expression profile on autocrine GH-mediated curcumin resistance, which was prevented by time-dependent curcumin treatment in T47D breast cancer cells. Dose- and time-dependent effect of curcumin on T47D wt and GH+ breast cancer cells were evaluated by MTT cell viability and trypan blue assay. Apoptotic effect of curcumin was determined by PI and Annexin V/PI FACS flow analysis. Immunoblotting performed to investigate the effect of curcumin on PI3K/Akt/MAPK, NF-κB signaling. miR182-96-183 cluster expression profile was observed by qRT-PCR. Overexpression of GH triggered resistant profile against curcumin (20 µM) treatment for 24 h, but this resistance was accomplished following 48 h curcumin exposure. Concomitantly, forced GH induced invasion and metastasis through EMT and NF-κB activation were prevented by long-term curcumin exposure in T47D cells. Moreover, 48 h curcumin treatment prevented the autocrine GH-mediated miR-182-96-183 cluster expression stimulation in T47D cells. In consequence, curcumin treatment for 48 h, prevented autocrine GH-triggered invasion-metastasis, EMT activation through inhibiting NF-κB signaling and miR-182-96-183 cluster expression and induced apoptotic cell death by modulating Bcl-2 family members in T47D breast cancer cells.

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Abbreviations

BRMS1L:

Breast cancer metastasis suppressor-1 like

DAPI:

4′,6-Diamidino-2-phenylindole

DiOC6:

3,3′-Dihexyloxacarbocyanine iodide

DMSO:

Dimethylsulfoxide

EMT:

Epithelial to mesenchymal transition

GH:

Growth hormone

GHR:

GH receptor

miRNAs:

microRNAs

MMP:

Matrix metalloproteinase

NF-κB:

Nuclear factor kappa B

PI:

Propidium iodide

PVDF:

Polyvildifluoride

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

STAT-5:

Signal transducer and activator of transcription 5

TBS:

Tris-buffered saline

TIMP:

Tissue inhibitory matrix proteinase

VEGF:

Vascular endothelial growth factor

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Acknowledgements

This work was supported by The Scientific and Technological Research Council of TURKEY (TUBITAK), Grand Number: 113Z791 and Istanbul Kultur University Scientific Projects Support Center. Authors wish to thank Busra Alper for her technical support in some immunoblotting results in Figs. 1d, 3a, 4a, 5a.

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Authors and Affiliations

  1. Molecular Biology and Genetics Department, Science and Letters Faculty, Istanbul Kultur University, Atakoy Campus, 34156, Istanbul, Turkey

    Ajda Coker-Gurkan, Derya Bulut, Recep Genc, Elif-Damla Arisan, Pınar Obakan-Yerlikaya & Narcin Palavan-Unsal

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  1. Ajda Coker-Gurkan
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  2. Derya Bulut
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Correspondence to Ajda Coker-Gurkan.

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This article does not involve any studies with human participants or animals performed by any of the authors. The research has been performed on commercially available cell lines.

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11033_2018_4479_MOESM1_ESM.jpg

The growth hormone expression profile in T47D GH+ breast cancer cell. The time dependent extracellular expression of GH in T47D breast cancer cell after GH inserted pcDNA3.1 (+) vector transfection and neomycin selection was determined by A. Immunoblotting and B. GH ELISA. P: Pellet, M: Medium, Humotrop was used as a positive control and β-actin was selected as a loading control.

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Coker-Gurkan, A., Bulut, D., Genc, R. et al. Curcumin prevented human autocrine growth hormone (GH) signaling mediated NF-κB activation and miR-183-96-182 cluster stimulated epithelial mesenchymal transition in T47D breast cancer cells. Mol Biol Rep 46, 355–369 (2019). https://doi.org/10.1007/s11033-018-4479-y

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