Camel Urine Promotes Sensitization to Doxorubicin by Inhibiting Epithelial-Mesenchymal Transition and Modulating NF-κB-Snail Signaling Pathway in Breast Cancer Cells.

Mashael Al-Mutairi,Suad Alfadli,Ibtehal Matar,Awated Al-Mutairi

doi:10.31557/apjcp.2021.22.12.4017

Mashael Al-Mutairi, Suad Alfadli + Show 2 more

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https://doi.org/10.31557/apjcp.2021.22.12.4017

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Abstract

Background:Camel urine (CU) has been used as traditional treatment in the Arabian Peninsula for centuries. Although, researchers have reported CU anti-cancer effects, the exact mechanism(s) of action involved has not been fully elucidated. The epithelial–mesenchymal transition EMT is a phenotypic switch that promotes the acquisition of a fibroblastoid-like morphology by epithelial tumor cells, resulting in enhanced tumor cell motility and invasiveness. EMT has been shown to contribute to metastasis and chemoresistance of carcinomas. For that, in the present study, we have assessed the potential mechanism (s) by which CU exert its anti-cancer effects and its possible synergistic therapeutic effect with Doxorubicin (DOX) in breast cancer cells. Methods:Determination of anti-proliferative and apoptosis validation of CU was performed by 3-(4,5-Dimethylthiazol-2-yl)-2,5,-diphenyltetrazolium bromide (MTT), annexin-V-fluorescein isothiocyanate assays, and Western blot. EMT protein markers, migration and invasion of cells were determined by Western blot or immunofluorescent staining, Scratch assay, Transwell invasion assay, respectively. Results:CU applied a significant anti-cancer effect on breast cancer cells via induction of DNA damage and apoptosis in a concentration- and time-dependent manner. Also, CU remarkably reversed the EMT by downregulating N-cadherin and Vimentin expression and upregulating E-cadherin expression. As a result, the stemness, migration and invasion of breast cancer cells were also inhibited, which was likely mediated by NF-κB-Snail signalling pathway and its downstream inflammatory effectors. CU successfully enhanced DOX cytotoxicity by reversing EMT which possibly through inhibition of NF-κB-Snail signalling and subsequently inflammation. Thus, our study provides new mechanistic bases for the therapeutic application of CU that may improve the outcomes of anti-cancer chemotherapy.

Highlights

Breast cancer is the most common cancer affecting women worldwide (Siegel et al, 2020)
Thousands of new cases are identified each year and an estimated 2.3 million new cases, representing 11.7% of all cancer cases and 685,000 deaths reported in 2020 (Sung et al, 2020). Conventional treatment regimens such as chemotherapy can suppress the initial phases of breast cancer development, the high morbidly and mortality rate of the disease is dependent on its ability to metastasize (Chaffer and Weinberg, 2011; Bai et al, 2016)
Immunostaining showed a clear increase in p-γ-H2AX expression in the nucleus after treatment with Camel urine (CU) at a concentration as low as 10 mg/ ml for 24 hrs compared to untreated cells. These results indicate that CU decreases the viability of breast cancer cells by stimulating apoptosis process

Summary

Introduction

Breast cancer is the most common cancer affecting women worldwide (Siegel et al, 2020). EMT, followed by polarity loss and motility gain, has been shown to play a crucial role in the metastasis process (Brabletz, 2012; Hou et al, 2014) Epithelial protein markers such as E-cadherin that typically encourage cell-cell contact may be lost during EMT, while cells acquire mesenchymal protein markers such as N-cadherin and Vimentin to boost their ability to migrate and invade. Both steps are crucial events in the initial phase of metastasis (Polyak and Weinberg, 2009; Thiery et al, 2009). Our study provides new mechanistic bases for the therapeutic application of CU that may improve the outcomes of anti-cancer chemotherapy

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