Islet Co-Expression of CD133 and ABCB5 in Human Retinoblastoma Specimens

Abstract

Background The role of CD133 und ABCB5 is discussed in treatment resistance in several types of cancer. The objective of this study was to evaluate whether CD133⁺/ABCB5⁺ colocalization differs in untreated, in beam radiation treated, and in chemotherapy treated retinoblastoma specimens. Additionally, CD133, ABCB5, sphingosine kinase 1, and sphingosine kinase 2 gene expression was analyzed in WERI-RB1 (WERI RB1) and etoposide-resistant WERI RB1 subclones (WERI ETOR).

Methods Active human untreated retinoblastoma specimens (n = 12), active human retinoblastoma specimens pretreated with beam radiation before enucleation (n = 8), and active human retinoblastoma specimens pretreated with chemotherapy before enucleation (n = 7) were investigated for localization and expression of CD133 and ABCB5 by immunohistochemistry. Only specimens with IIRC D, but not E, were included in this study. Furthermore, WERI RB1 and WERI ETOR cell lines were analyzed for CD133, ABCB5, sphingosine kinase 1, and sphingosine kinase 2 by the real-time polymerase chain reaction (RT-PCR).

Results Immunohistochemical analysis revealed the same amount of CD133⁺/ABCB5⁺ colocalization islets in untreated and treated human retinoblastoma specimens. Quantitative RT-PCR analysis showed a statistically significant upregulation of CD133 in WERI ETOR (p = 0.002). No ABCB5 expression was detected in WERI RB1 and WERI ETOR. On the other hand, SPHK1 (p = 0.0027) and SPHK2 (p = 0.017) showed significant downregulation in WERI ETOR compared to WERI RB1.

Conclusions CD133⁺/ABCB5⁺ co-localization islets were noted in untreated and treated human retinoblastoma specimens. Therefore, we assume that CD133⁺/ABCB5⁺ islets might play a role in retinoblastoma genesis, but not in retinoblastoma treatment resistance.

Zusammenfassung

Hintergrund Die Rolle von CD133 und ABCB5 in der Therapieresistenz von verschiedenen Tumoren wird aktuell diskutiert. Ziel der Studie war es zu untersuchen, ob Unterschiede in der CD133⁺/ABCB5⁺-Kolokalisation bei bestrahlten, mit Chemotherapie behandelten und nicht vorbehandelten Retinoblastomproben bestehen. Zusätzlich wurde die Genexpression von CD133, ABCB5, Sphingosinkinase 1 und Sphingosinkinase 2 in der WERI-RB1-Zelllinie und seinem Etoposid-resistenten Subklon (WERI-ETOR) analysiert.

Methoden Humane unbehandelte (n = 12), mit Bestrahlung vorbehandelte (n = 8) und mit Chemotherapie vorbehandelte (n = 7) enukleierte Augen mit aktivem Retinoblastomgewebe (IIRC D, jedoch nicht E) wurden mittels immunhistologischer Untersuchung auf die Lokalisation und Expression von CD133 und ABCB5 hin analysiert. Weiter wurden die Zelllinien WERI-RB1 und WERI-ETOR mittels Real-Time Polymerase Chain Reaction (RT-PCR) auf die Expression von CD133, ABCB5, Sphingosinkinase 1 und Sphingosinkinase 2 untersucht.

Ergebnisse Die immunohistochemische Analyse zeigte ähnliche Mengen an CD133⁺/ABCB5⁺-Kolokalisationsinseln in behandelten und unbehandelten humanen Retinoblastomproben. Die quantitative RT-PCR-Analyse zeigte eine signifikante Hochregulation von CD133 in WERI-ETOR (p = 0,002). Eine ABCB5-Expression konnte weder in WERI-RB1 noch in WERI-ETOR nachgewiesen werden. Sphingosinkinase 1 (p = 0,0027) und Sphingosinkinase 2 (p = 0,017) waren in WERI-ETOR signifikant herunterreguliert.

Schlussfolgerung CD133⁺/ABCB5⁺-Kolokalisationsinseln konnten in behandelten und unbehandelten humanen Retinoblastomproben nachgewiesen werden. Wir schließen daraus, dass CD133⁺/ABCB5⁺-Zellen eventuell eine Rolle in der Retinoblastomgenese spielen können, aber wohl keinen Einfluss auf die Resistenzentwicklung im Retinoblastom haben.

Key words

ocular tumors - retina - pediatric ophthalmology

Schlüsselwörter

okuläre Tumoren - Retina - Kinderophthalmologie

Publication History

Received: 04 September 2020

Accepted: 07 June 2021

Article published online:
27 September 2021

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