Naslov
Specific and overlapping GLI transcription targets in melanoma

Autori
Rinčić, Nikolina ; Bartoniček, Nenad ; Kurtović, Matea ; Musani, Vesna ; Trnski, Diana ; Ozretić, Petar ; Sabol, Maja

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, neobjavljeni rad, znanstveni

Skup
EACR 2021 : : Innovative Cancer Science

Mjesto i datum
Online, 09.06.2021. - 12.06.2021

Vrsta sudjelovanja
Poster

Vrsta recenzije
Podatak o recenziji nije dostupan

Ključne riječi
Melanoma ; GLI proteins ; ChIP-seq ; BRAF ; NRAS

Sažetak
Hedgehog-GLI signaling pathway is one of the key regulators of normal development. Its aberrant activation has been linked to several types of cancer, including melanoma. Previous studies in our laboratory demonstrated that BRAF and NRAS melanomas have different response to HH-GLI signaling pathway inhibition. Identifying GLI protein functions in melanoma cell lines with different mutational background represents a distinct potential for the development of combined therapy with HH-GLI signaling pathway and BRAF/NRAS inhibitors. For that purpose, chromatin immunoprecipitation sequencing (ChIP-seq) was performed. Firstly, 14 melanoma cell lines were collected and their mutation status was confirmed by Sanger sequencing. The cell lines were divided into three categories based on their mutational background: BRAF-mutated, NRAS-mutated, and wild- type for BRAF and NRAS. To enhance the ChIP-seq performance we selected one cell line from each group with the strongest protein expression of HH- GLI signaling pathway components. After all, three cell lines were selected, CHL-1 for the wild-type cell group, A375 for the cell group with BRAF gene mutation and MEL224 for the cell group with NRAS gene mutation. The selected cell lines were double- cross linked, fixed DNA-protein complexes were enzymatically fragmented and the desired chromatin fragments were immunoprecipitated using GLI protein specific antibodies. The purified chromatin samples were used for library preparation and proceeded for next- generation sequencing (NGS). The target genes analysis will provide new insights into unique and overlapping functions of GLI proteins and potentially lead to novel therapeutic perspectives in BRAF/NRAS mutated melanoma.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)

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