Vsebnost in vloga kemokina CCL5 in njegovega receptorja CCR5 v glioblastomu multiforme

Hrastar, Barbara (author), Lah Turnšek, Tamara (mentor), Novak, Metka (co-mentor)

Glioblastom multiforme (GB) je najbolj pogost in najbolj agresiven možganski tumor pri človeku, ki ga opredeljuje visoka stopnja proliferacije in invazije v možgansko tkivo. Zdravljenje je zaradi nepopolne kirurške odstranitve in odpornosti tumorja na radio- in kemoterapijo neuspešno, poleg tega pa je ponovitev GB zelo pogosta. Kemotaktični citokini, oz. kemokini so vpleteni v številne signalne poti, tako v fizioloških pogojih kot tudi v bolezenskih stanjih. Vsebnost CCL5 in CCR5 smo določali na nivoju genskega izražanja z metodo qPCR-RT v GB tkivnih vzorcih bolnikov in primarnih celičnih linijah GB ter na proteinskem nivoju z imunohistokemijsko analizo. CCL5 in CCR5 sta povišano izražena v GB vzorcih bolnikov ter v ponovljenih GB, kar je povezano s slabšo prognozo. Kemokin CCL5 in njegov receptor CCR5 v celicah GB vplivata na njihovo proliferacijo, preživetje in invazijo. Kemotaktično gibanje celic v smeri gradienta CCL5/CCR5 lahko blokiramo z antagonistom receptorja CCR5, Maravirocom (MVR), s katerim smo preverili vpliv inhibicije na preživetje in invazijo celic GB. Z uporabo MTT testa smo pokazali, da MVR nima vpliva na celično preživetje. Z 2D modelom celične invazije pa smo dokazali inhibitorni vpliv MVR na invazijo celic GB, ob prisotnosti signala CCL5.

• Language: Slovenian
• Keywords: biologija, glioblastom multiforme, kemokini, Maraviroc
• Work type: Master's thesis/paper
• Organization: BF - Biotechnical Faculty
• Year: 2019
• PID: 20.500.12556/RUL-110092
• COBISS.SI-ID: 5250895
• Publication date in RUL: 12.09.2019

Secondary language

• Language: English
• Title: Expression and significance of chemokine ligand CCL5 and its receptor CCR5 in glioblastoma

Abstract

Glioblastoma multiforme (GB) is the most frequent and aggressive brain tumor in human, with high proliferative and diffuse invasive properties. The outcome of the disease is dismal, due to impossible total surgical resection and tumor resistance to irradiation and chemotherapy. Moreover, the tumor often recurs in a period of a few months. Chemotactic cytokines or chemokines are involved in many signaling pathways within physiological homeostasis and in pathologic states. In GB tissues samples from patients and in GB cells we have analyzed CCL5 and CCR5 gene expression using qPCR-RT and CCL5 and CCR5 protein expression using immunohistochemistry. Both CCL5 and CCR5 expressions were increased in GB tissue samples and in recurrent GB tissues, which means very bad prognosis for survival. Chemokine CCL5 and its receptor CCR5 in GB cells are involved in proliferation, cell survival and invasion. We can block CCL5 induced chemotaxis with CCR5 antagonist Maraviroc (MVR). Using this inhibitor, we verified the inhibitory effect on cell survival and invasion of GB cells, when challenged by CCL5. Using MTT assay, we also demonstrated that MVR had no effect on GB cell viability.

• Keywords: biology, glioblastoma multiforme, chemokines, Maraviroc