Pivotal Role of AKT2 during Dynamic Phenotypic Change of Breast Cancer Stem Cells.

Gener Gener,Abasolo Abasolo,Arango Arango,Díaz-Riascos Díaz-Riascos,Perez Perez,Schwartz Schwartz,Fernández Fernández,Pindado Pindado,Rafael Rafael,Casas Casas,Seras-Franzoso Seras-Franzoso

doi:10.3390/cancers11081058

Abstract

Therapeutic resistance seen in aggressive forms of breast cancer remains challenging for current treatments. More than half of the patients suffer from a disease relapse, most of them with distant metastases. Cancer maintenance, resistance to therapy, and metastatic disease seem to be sustained by the presence of cancer stem cells (CSC) within a tumor. The difficulty in targeting this subpopulation derives from their dynamic interconversion process, where CSC can differentiate to non-CSC, which in turn de-differentiate into cells with CSC properties. Using fluorescent CSC models driven by the expression of ALDH1A 1(aldehyde dehydrogenase 1A1), we confirmed this dynamic phenotypic change in MDA-MB-231 breast cancer cells and to identify Serine/Threonine Kinase 2 (AKT2) as an important player in the process. To confirm the central role of AKT2, we silenced AKT2 expression via small interfering RNA and using a chemical inhibitor (CCT128930), in both CSC and non-CSC from different cancer cell lines. Our results revealed that AKT2 inhibition effectively prevents non-CSC reversion through mesenchymal to epithelial transition, reducing invasion and colony formation ability of both, non-CSC and CSC. Further, AKT2 inhibition reduced CSC survival in low attachment conditions. Interestingly, in orthotopic tumor mouse models, high expression levels of AKT2 were detected in circulating tumor cells (CTC). These findings suggest AKT2 as a promising target for future anti-cancer therapies at three important levels: (i) Epithelial-to-mesenchymal transition (EMT) reversion and maintenance of CSC subpopulation in primary tumors, (ii) reduction of CTC and the likelihood of metastatic spread, and (iii) prevention of tumor recurrence through inhibition of CSC tumorigenic and metastatic potential.

Highlights

Drug resistance in breast cancer cell populations is acquired through phenotypic changes mediated by either epithelial-to-mesenchymal transition (EMT), or tumor cell drift towards a cancer stem cell (CSC) phenotype [1,2]
Of the tdTomato subpopulation (Figure 1C), which accounted for 14.7% ± 2.8% of tdTomato cells within tdTomato+ cells within the MDA-MB-231-ALDH1A1/tdTomato cell line (Figure 1B)
The increased expression of EMT genes (AKT2, TWIST, and mTOR) in tdTomato− tumors, in circulating tumor cells (CTC) and in mammospheres, highlights the importance of AKT2 in tumor growth and cancer stem cells (CSC) malignancy. To pursue this line of investigation further, we explored the consequences of AKT2 silencing in three different cancer cell lines, and their CSC-like fractions (MDA-MB-231, HCT8, and MCF7)

Summary

Introduction

Drug resistance in breast cancer cell populations is acquired through phenotypic changes mediated by either epithelial-to-mesenchymal transition (EMT), or tumor cell drift towards a cancer stem cell (CSC) phenotype [1,2]. Cancers 2019, 11, 1058 cancer treatment, tumors contain a heterogeneous mixture of various cell types, with different stem-like and mesenchymal phenotypes. In this intricate network made of diverse cell populations, the characteristics of mesenchymal and stem-like phenotype are often intertwined, becoming difficult to distinguish from one another [2,3,4,5]. EMT cells, as well as CSC, are described as responsible for metastatic spread, tumor progression, and resistance. The EMT conversion stimulates expression of CSC genes and vice-versa, CSC often expresses genes characteristic for mesenchymal cells [1,2,6]

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Results

Conclusion