Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F17%3A00067236" target="_blank" >RIV/00159816:_____/17:00067236 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14110/17:00095052 RIV/62156489:43210/17:43911858 RIV/00216305:26620/17:PU124934

Výsledek na webu

<a href="http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path" target="_blank" >http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.18632/oncotarget.19914" target="_blank" >10.18632/oncotarget.19914</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations

Popis výsledku v původním jazyce

In this study, we describe the establishment of the human papillomavirus 18-positive, stage II, grade 1, T2N0M0 head and neck tumor primary cell line derived from oral squamous cell carcinoma of a non-smoking patient by using two different protocols. Furthermore, a preparation of subpopulations derived from this primary cell line according to the cluster of differentiation molecules CD44/CD90 status using magnetic bead-based separation and their characterization was performed. Impedance-based real-time cell analysis, enzyme-linked immunsorbant assay (ELISA), wound-healing assay, flow-cytometry, gene expression analysis, and MTT assay were used to characterize these four subpopulations (CD44(+)/CD90(-), CD44(-)/CD90(-), CD44(+)/CD90(+), CD44(-)/CD90(-)). We optimised methodics for establishement of primary cell lines derived from oral squamous cell carcinoma tissue samples and subsequent separation of mesenchymal (CD90(+)) and epithelial (CD90(-)) types of tumorous cells. Primary cell line prepared by using trypsin proteolysis was more viable than the one prepared by using collagenase. According to our results, CD90 separation is a necessary step in preparation of permanent tumor-tissue derived cell lines. Based on the wound-healing assay, CD44(+) cells exhibited stronger migratory capacity than CD44(-) subpopulations. CD44(+) subpopulations had also significantly higher expression of BIRC5 and SOX2, lower expression of FLT1 and IL6, and higher levels of basal autophagy compared to CD44(-) subpopulations. Furthermore, co-cultivation experiments revealed that CD44(-)/CD90(+) cells supported growth of epithelial tumor cells (CD44(+)/CD90(-)). On the contrary, factors released by CD44(+)/CD90(+) type of cells seem to have rather inhibiting effect. The most cisplatin-resistant subpopulation with the shortest doubling time was CD44(-)/CD90(+), but this subpopulation had a low migratory capacity.

Název v anglickém jazyce

Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations

Popis výsledku anglicky

In this study, we describe the establishment of the human papillomavirus 18-positive, stage II, grade 1, T2N0M0 head and neck tumor primary cell line derived from oral squamous cell carcinoma of a non-smoking patient by using two different protocols. Furthermore, a preparation of subpopulations derived from this primary cell line according to the cluster of differentiation molecules CD44/CD90 status using magnetic bead-based separation and their characterization was performed. Impedance-based real-time cell analysis, enzyme-linked immunsorbant assay (ELISA), wound-healing assay, flow-cytometry, gene expression analysis, and MTT assay were used to characterize these four subpopulations (CD44(+)/CD90(-), CD44(-)/CD90(-), CD44(+)/CD90(+), CD44(-)/CD90(-)). We optimised methodics for establishement of primary cell lines derived from oral squamous cell carcinoma tissue samples and subsequent separation of mesenchymal (CD90(+)) and epithelial (CD90(-)) types of tumorous cells. Primary cell line prepared by using trypsin proteolysis was more viable than the one prepared by using collagenase. According to our results, CD90 separation is a necessary step in preparation of permanent tumor-tissue derived cell lines. Based on the wound-healing assay, CD44(+) cells exhibited stronger migratory capacity than CD44(-) subpopulations. CD44(+) subpopulations had also significantly higher expression of BIRC5 and SOX2, lower expression of FLT1 and IL6, and higher levels of basal autophagy compared to CD44(-) subpopulations. Furthermore, co-cultivation experiments revealed that CD44(-)/CD90(+) cells supported growth of epithelial tumor cells (CD44(+)/CD90(-)). On the contrary, factors released by CD44(+)/CD90(+) type of cells seem to have rather inhibiting effect. The most cisplatin-resistant subpopulation with the shortest doubling time was CD44(-)/CD90(+), but this subpopulation had a low migratory capacity.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30204 - Oncology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Oncotarget

ISSN

1949-2553

e-ISSN

—

Svazek periodika

8

Číslo periodika v rámci svazku

39

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

16

Strana od-do

66254-66269

Kód UT WoS článku

000410291200128

EID výsledku v databázi Scopus

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