Cancer cell drug resistance develops via several mechanisms, including apoptosis evading, drug efflux, cell reprogramming, DNA repair. Recently numerous data pointed out an ability of cancer cells reversibly exit from cell cycle to enter in G0 phase that may favor cancer cell survival. Therefore, the goal of the study was to determine transcriptional phenotype associated with quiescent/senescent state acquisition. Melanoma cells were treated by Dacarbazine to induce their transition in G0 phase of a cell cycle followed by Ki-67 and flow cytometry-based cell cycle analysis. Beta-galactosidase expression was identified by hydrolysis of 5-bromo-4-chloro-3-indolyl β-D-galactopyranoside resulting in blue staining of beta-galactosidase positive cells. Microarray was performed to access transcriptional profiling alterations. Cell adhesion capacities of G0-positive cells were determined by centrifugal force. Indeed, 1.2 mmol Dacarbazine induced a 4-fold increase in the percentage of G0-positive BRO and SK-MEL-2 cells that was also visualized by Ki-67 immunostaining. Opposite, the percentage of beta-galactosidase-positive cells was increased in SK-MEL-2 from 0.3 to 0.7% and did not altered in BRO cells. According to transcriptomic study the top signal pathways enriched among dysregulated genes were «Cell cycle», «p53 transcriptional gene network», «Focal adhesion». Next, a cell adhesion assay was performed in melanoma cells following treatment with Dacarbazine. Both BRO and SK-MEL-2 cells exhibited an elevated percentage of G0-positive cells among the adhesive cells. In conclusion, human melanoma cells may enter to G0 under Dacarbazine treatment that results in significant alterations in transcriptional profile. Besides, G0-positive melanoma cells demonstrated enhanced adhesive capacities that may be related to a pre-metastatic niche formation and cancer cell dissemination.

ESDR492 - Altered focal adhesion pattern of G0-positive melanoma cells

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