Naslov
Knockdown of the integrin subunit alpha V in MDA-MB-435S cells decreases sensitivity to cisplatin

Autori
Sedda, Delphine

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, diplomski rad, diplomski

Fakultet
Masters "Science of Life", Master thesis of Biochemistry, Molecular Biology, Biotechnology

Mjesto
Orleans

Datum
02.09

Godina
2016

Stranica
40

Mentor
Ambriović Ristov, Andreja

Ključne riječi
integrin alpha V beta 3; integrin alpha V beta 5; siRNA; triple negative breast cancer; melanoma; cisplatin

Sažetak
Triple Negative Breast Cancer (TNBC) and melanoma are two aggressive cancers with a poor prognosis. Integrins are heterodimeric transmembrane proteins that mediate the adhesion of cells between them and extracellular matrix. They regulate proliferation, migration, metastasis, angiogenesis, and also cell sensitivity to antitumor drugs. It has been shown in MDA-MB-435S cells that integrin subunit alpha V silencing using integrin-specific siRNA increases sensitivity of cells to paclitaxel but reduces sensitivity to cisplatin. The origin of this cell line, TNBC or melanoma, has been a matter of debate. However, this work is part of the project in which the results obtained in this cell line will be compared to those obtained in different TNBC and melanoma cell lines to evaluate the potential of integrin silencing in sensitisation to chemotherapy. In this study we investigated the mechanism behind the decreased sensitivity of MDA-MB-435S cells to cisplatin after subunit alpha V knockdown. Our results showed a comparable kinetics of DNA-double strand breaks, with or without integrin alpha V knockdown, as judged by phosphorylation of H2AX. We showed that ERK pathway is not involved in decrease of cisplatin sensitivity in MDA-MB-435S after integrin subunit alpha V knockdown. We also observed decreased expression of pFAK and pAkt upon integrin alpha V knockdown indicating that they belong to the signal pathway triggered by integrin heterodimers alpha V beta 3 and/or alpha V beta 5. In addition, upon cisplatin exposure we observed the abrogation of activation of pAkt in cells upon integrin alpha V knockdown. However, we showed that Akt pathway is not involved in decreased sensitivity of MDA-MB-435S cells to cisplatin upon integrin alpha V knockdown. We isolated stably transfected MDA-MB-435S cell clones by transfection with plasmid harbouring integrin alpha V-specific shRNA in which we showed decreased expression of integrins alpha V beta 3 and alpha V beta 5 and decreased sensitivity to cisplatin, thus confirming the results of transient transfection with integrin alpha V specific siRNA. These cell clones will facilitate the studies of the signalling pathways involved in the modulation of sensitivity to drugs exposure after integrin alpha V knockdown.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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