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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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.
KeywordsBreast cancer NF-κB Epithelial mesenchymal transition miRNA
Breast cancer metastasis suppressor-1 like
Epithelial to mesenchymal transition
Nuclear factor kappa B
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
Signal transducer and activator of transcription 5
Tissue inhibitory matrix proteinase
Vascular endothelial growth factor
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.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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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