Reprogramming human A375 amelanotic melanoma cells by catalase overexpression: Reversion or promotion of malignancy by inducing melanogenesis or metastasis

Abstract

Advanced melanoma is the most aggressive form of skin cancer. It is highly metastatic and dysfunctional in melanogenesis; two processes that are induced by H2O2. This work presents a melanoma cell model with low levels of H2O2 induced by catalase overexpression to study differentiation/dedifferentiation processes. Three clones (A7, C10 and G10) of human A375 amelanotic melanoma cells with quite distinct phenotypes were obtained. These clones faced H2O2 scavenging by two main strategies. One developed by clone G10 where ROS increased. This resulted in G10 migration and metastasis associated with the increased of cofilin-1 and CAP1. The other strategy was observed in clone A7 and C10, where ROS levels were maintained reversing malignant features. Particularly, C10 was not tumorigenic, while A7 reversed the amelanotic phenotype by increasing melanin content and melanocytic differentiation markers. These clones allowed the study of potential differentiation and migration markers and its association with ROS levels in vitro and in vivo, providing a new melanoma model with different degree of malignancy.

Cofilin-1 levels and intracellular localization are associated with melanoma prognosis in a cohort of patients

“…In this sense, we demonstrated an association between cofilin-1 levels and malignancy, measured by Breslow thickness, melanoma staging, mitotic index and presence of metastasis, in melanoma patients. This is in agreement with an in vitro previous study, where we reported high levels of cofilin-1 in migrating cells with metastatic ability compared with non-migratory neither metastatic control cells in an experimental model of human melanoma [ 22 ]. Besides, proteomic studies showed differential expression of cofilin-1 isoforms in two melanoma cell lines compared with a melanocyte cell line [ 31 ] and increased expression of cofilin-1 in metastatic lymph node compared with matched human primary cutaneous melanoma tissue of the same patients [ 32 ].…”

“…The intracellular localization of cofilin-1 was evaluated in a cellular model of melanoma developed in our laboratory [ 22 ] and in tissue sections of benign and malignant melanocytic lesions from patients. Immunocytofluorescence assay of cofilin-1 in non-metastatic A375 and metastatic A375-G10 cells was performed as previously described [ 22 ]. For immunohistofluorescence in patient samples, tissue sections were deparaffinized and rehydrated as described above.…”

“…In this regard, cofilin-1 overexpression was associated with increased migration in colon adenocarcinoma cells [ 19 ] and has been used as poor prognosis marker, related to metastasis, in non-small cell lung cancer patients [ 20 , 21 ] and invasive breast cancer cells [ 17 ]. Besides, a previous work from our laboratory demonstrated in a melanoma model with different degree of malignancy a correlation among the high levels of cofilin-1 and the migratory, invasive and metastatic ability of cells [ 22 ], suggesting its relevance in melanoma progression.…”

Comparison of Xiphophorus and human melanoma transcriptomes reveals conserved pathway interactions

“…We used relatively high xmrk expression levels to hallmark a dedifferentiation state of Xiphophorus melanoma cells. Melanoma stem cell makers and metastasis‐related genes prom1 (CD133), itga6, itgab, itgb1, tbx2 , zeb1 , cdh7 , cdh20, and cfl (Figure c; (Argaw‐Denboba et al., ; Bosserhoff, Ellmann, & Kuphal, ; Bracalente et al., ; Madjd et al., ; Moore et al., ; Zhao et al., ; Zimmerer et al., ) were found to be co‐expressed with xmrk . These observations show xmrk is associated with a cluster of genes that are capable of maintaining the cells in an undifferentiated state.…”

“…This feature is derived from neural crest progenitor cells that respond to morphogenetic cues from tissue microenvironments and give rise to respective lineages, including melanocytes (Simoes-Costa & Bronner, 2015; Takahashi, Sipp, & Enomoto, 2013). Studies have revealed that melanoma is organized and driven by a subpopulation of cancer cells that have the properties of dedifferentiated stem cells, such as disruption of dendricity, enhanced cell proliferation, and loss of pigmentation (Bracalente et al, 2016; Frank, Schatton, & Frank, 2010; Kalluri & Weinberg, 2009; Lee & Vasioukhin, 2008; Royer & Lu, 2011; Saez-Ayala et al, 2013; Schiaffino, 2010; Serafino et al, 2004). The xmrk oncogene can induce transformation of differentiated melanocytes (Delfgaauw et al, 2003; Wellbrock et al, 1998; Wellbrock et al, 2002).…”