engleski

Newly established ovarian cancer model of acquired carboplatin resistance reveals new targets to fight drug resistance and metastasis

Despite the notable progress in the development of new technologies and methods for early tumor diagnostic, ovarian cancer is still the deadliest gynecological disease worldwide. Lack of early- stage symptoms and specific biomarkers results in late-stage diagnosis when cancer already spread. Even a small number of the initially responsive ovarian cancer patients usually develop drug resistance upon therapy. Here, we present an ab ovo ovarian cancer model to study acquired resistance to carboplatin (CBP) and CBP-induced epithelial-mesenchymal transition (EMT). Parental MES-OV cells were repeatedly treated with increasing doses of CBP. Resistant variants, once stabile, were characterized in terms of carboplatin and paclitaxel (TAX) response by resazurin survival assay, migratory potential by scratch assay, and invasive potential by the transwell assay. Expressions of EMT markers were checked by RT-qPCR. Transcriptome analysis was performed on all established variants. The data were analyzed by two different approaches, the first one based on the differentially expressed genes (DEG), and the other one focusing on the genes that correlate with sensitivity to CBP (CBP- correlating genes, CCORG). Functional and clinical roles of selected candidates were examined by siRNA transfections in vitro and using bioinformatics tools in silico, respectively. The roles of enriched pathways were checked by survival assays on cells pre-treated with a set of specific inhibitors. Gradual increase in CBP and TAX resistance, as well as an increase in migration and invasion rates, were reported in CBP resistant cell lines. Two approaches in data analysis resulted in groups of completely different genes highlighting different signaling pathways. Functional analysis revealed that HES7 probably plays a role in the development of CBP resistance, while SERPINE2 probably plays a role in metastases. TMEM200A, PRKAR1B, and FBLN5 seem to be involved in both phenomena, while TMEM200A could also serve as a biomarker of resistance. The significance of enriched signaling pathways is yet to be determined. The characterized cellular model, along with two distinct approaches to transcriptome analysis, presents a novel platform in research of drug resistance and drug-induced EMT. They enabled the exploration of novel candidate genes, possible therapeutic targets, and biomarkers, and support the narrative of two phenomena being interconnected.

ovarian cancer ; carboplatin resistance ; CBP ; drug resistance ; metastasis

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