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Efficacy and mechanism of action of Deguelin in suppressing metastasis of 4T1 cells

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Abstract

Cancer related deaths in breast cancer patients are due to metastasis of the disease. Murine 4T1 cells (Murine mammary cancer cell line developed from 6-thioguanine resistant tumor) provide an excellent research tool for metastasis related studies because these cells are highly aggressive and readily metastasize to the lungs. In this study we determined the effect of Deguelin on in vivo/vitro growth and metastasis of 4T1 cells. Deguelin inhibited the in vitro growth of 4T1 cells in a time and dose dependent manner accompanied with reduced nuclear PCNA immunostaining. In cells treated with Deguelin, reduced expression of nuclear c-Met, and its downstream targets such p-ERK and p-AKT was observed. Deguelin reduced the cell migration in 4T1 cells as determined by scratch wound assay. Combined treatment with Deguelin + ERK or PI3K/AKT inhibitor had no additional effect on cell migration. These results indicated that the action of Deguelin on cell migration may be mediated by AKT and ERK mediated signaling pathways. In vivo, Deguelin treatment significantly inhibited growth of 4T1 cells. Deguelin also reduced the occurrence of metastatic lung lesions by 33 % when cells were injected intravenously into Balb/c female mice. There was no difference in the body weight, nor was there a difference in liver and spleen weights between vehicle treated-control and Deguelin-treated animals, which indicated that Deguelin was nontoxic at the dose used in the present study. These results provide rationale for developing Deguelin as a chemotherapeutic agent for triple negative breast cancer patients.

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Abbreviations

4T1:

Mouse mammary tumor cell line 4T1

MDA-MB-231:

Human breast cancer cell line MDA-MB-231

ER:

Estrogen receptor

PR:

Progesterone receptor

HER-2:

Human epidermal growth factor receptor-2

PCNA:

Proliferating cell nuclear antigen

c-Met:

MET or MNNG HOS transforming gene

HGF:

Hepatocyte growth factor

ERK:

Extracellular-signal-regulated kinases

p-ERK:

Phosphorylated ERK

AKT:

Mouse strain Ak-thymoma (t)

p-AKT:

Phosphorylated AKT

COX-2:

Cyclooxyhenase-2

ERB-B2:

Avian erythroblastosis oncogene B

TNBC:

Triple negative breast cancer

XIAP:

X-linked inhibitor of apoptosis protein

hsp90:

Heat shock protein 90

RT:

Reverse transcriptase

PBS:

Phosphate buffered saline

DAPI:

4′,6-diamidino-2-phenylindole

PI:

Propidium iodide

BSA:

Bovine serum albumin

DAB:

Diaminobenzedene

AIN 76A:

American Institute of Nutrition 76A

PI3K:

Phosphatidylinositol 3-kinases

DMSO:

Dimethylsulfoxide

FAK:

Focal adhesion kinase-1

HIF1α:

Hypoxia inducing factor 1alpha

i.p.:

Intraperitoneally

i.v.:

Intravenous

NFκB:

Nuclear factor kappa-light-chain-enhancer of activated B cells

Ras:

Rat sarcoma

MAPK:

Mitogen activated protein kinase

PKC:

Protein kinase C

IKB:

Inhibitor of kappa B

IKK:

I kappa B

LEF1:

Lymphoid enhancer-binding factor-1

TCF:

T Cell Factor

Wnt:

Combination of integration 1 (int) and drosophila wingless (wg)

c-MYC:

Myelocytomatosis oncogene

VEGF:

Vascular endothelial growth factor

MMP-7:

Matrix metalloproteinase 7

uPA:

Urokinase-type plasminogen activator

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Acknowledgments

This work was supported by National Cancer Institute USPHS Grant CA140321.

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Authors and Affiliations

  1. Cancer Biology Division, IIT Research Institute, 10 West 35th Street, Chicago, IL, 60616, USA

    Rajeshwari R. Mehta, Harshadadevi Katta, Amit Kalra, Rutulkumar Patel, Akash Gupta, Fatouma Alimirah, Genoveva Murillo, Xinjian Peng, Miguel Muzzio & Rajendra G. Mehta

  2. Department of Chemistry, Illinois Institute of Technology, Chicago, IL, 60616, USA

    Aditya Unni

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  1. Rajeshwari R. Mehta
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Correspondence to Rajendra G. Mehta.

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Mehta, R.R., Katta, H., Kalra, A. et al. Efficacy and mechanism of action of Deguelin in suppressing metastasis of 4T1 cells. Clin Exp Metastasis 30, 855–866 (2013). https://doi.org/10.1007/s10585-013-9585-6

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