Knockdown of ZEB1 reverses cancer stem cell properties in prostate cancer cells

Abstract

Prostate cancer (PCa) is the second most diagnosed type of cancer in men worldwide. Advanced PCa is resistant to conventional therapies and high recurrence has been associated with high rates of metastasis. Cancer stem cells (CSCs) have been proposed to be responsible for this, due to their ability of self-renewal and differentiation into other cell types. Zinc finger E-box-binding homeobox 1 (ZEB1), a transcription factor involved in the regulation of epithelial-mesenchymal transition (EMT), has been associated with the activation of several mechanisms that lead to resistance to treatment. As recent evidence has shown that CSCs may originate from non-CSCs during EMT, it was hypothesized that knocking down ZEB1 expression in PCa cell lines could revert some properties associated with CSCs. Using lentiviraltransduction, ZEB1 expression was silenced in the PCa DU145 and LNCaP cell lines. The mRNA and protein expression levels of key canonical CSC markers (Krüppel-like factor 4, SOX2, CD44 and CD133) were determined using reverse transcription-quantitative PCR and western blot analysis, respectively. In addition, the colony forming ability of the ZEB1-knockdown cells was evaluated, and the type of colonies formed (holoclones, paraclones and meroclones) was also characterized. Finally, the ability to form prostatospheres was evaluated in vitro. It was found that in ZEB1-knockdown DU145 cells, the expression levels of CSC phenotype markers (CD44, CD133 and SOX2) were decreased compared with those in the control group. Furthermore, ZEB1-knockdown cells exhibited a lower ability to form prostatospheres and to generate colonies. In conclusion, stable silencing of ZEB1 reversed CSC properties in PCa cell lines. Since ZEB1 is associated with malignancy, therapy resistance and a CSC phenotype in PCa cell lines, targeting ZEB1 may be a key factor to eradicate CSCs and improve the prognosis of patients with advanced PCa.

SPARC Induces E-Cadherin Repression and Enhances Cell Migration through Integrin αvβ3 and the Transcription Factor ZEB1 in Prostate Cancer Cells

“…Although all the subpopulations present during EMT (epithelial, early EMT, intermediate EMT, late EMT, and complete mesenchymal) can show some degree of plasticity, the evidence suggests that hybrid or intermediate EMT populations are those showing the highest degree of plasticity and clonogenic capacity which, under in vivo conditions, can give rise to tumors with different subpopulations [ 5 , 8 ]. Consistent with this, recent results from our laboratory show that PC3 cells with silenced SPARC have lower clonogenic capacity, lower ability to form prostatospheres under non-adherent culture conditions, as well as a lower expression of the CD44 marker and the transcriptional factor SOX2 [ 9 ].…”

Specific N-cadherin–dependent pathways drive human breast cancer dormancy in bone marrow

“…There were negligible changes in the expression of metastatic-related genes-vimentin, twist, slug, β-catenin, and Zeb1 (Fig 5G). We follow up on Zeb 1 because of its role in maintaining CSCs (Pérez et al, 2021). This was addressed with CSC extracts, resulting in a faint double band in the scramble and knockdown cells ( MAPK pathway has been reported to regulate stem cell renewal (Niu et al, 2017).…”

Zinc Favors Triple-Negative Breast Cancer’s Microenvironment Modulation and Cell Plasticity

“…In addition, we saw that the CD44(+)CD24(−/low) subpopulation was enriched in zinc, with CD44(+) as the specific marker associated with higher zinc content. In this context, it has been reported that ZEB1 regulates the CSC population of prostate cancer, also characterized by CD44 expression [35]. Interestingly, the presence of zinc reduced the ALDH marker in the tumorspheres of MDA-MB-231 and BrM2 cell lines.…”

“…We wanted to further study stemness in our cell model. Therefore, we examined whether there was a correlation between zinc content and the expression levels of the well-known stem-cell markers Oct4, Sox2, and Nanog [34], as well as other zinc-dependent tumorigenic markers, such as ZEB1 [35] and SerpinB2 [32]. Even though we observed some interesting tendencies in MDA-MB-231 cells, no significant differences were obtained for any marker (Supplementary Materials Figure S2).…”