Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 1.736
5-Year Impact Factor – 2.135
Index Copernicus  – 168.52
MEiN – 70 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2011, vol. 20, nr 3, May-June, p. 285–294

Publication type: original article

Language: English

Expression of Cell Vault Components MVP, TEP1 and vPARP in Cancerous Ovarian Tissues

Ekspresja składników krypt komórkowych MVP, TEP1 oraz vPARP w tkankach nowotworowych z jajnika

Witold Szaflarski1,, Patrycja Sujka-Kordowska1,, Radosław Januchowski1,, Karolina Jaszczyńska-Nowinka2,, Małgorzata Andrzejewska1,, Adnan Savan3,, Piotr Dzięgiel1,4,, Michał Nowicki1,, Maciej Zabel1,4,

1 Department of Histology and Embryology, Poznań University of Medical Sciences, Poland

2 Department of Gynecological Oncology, Poznań University of Medical Sciences, Poland

3 Department of Biomaterials and Experimental Dentistry, Poznań University of Medical Sciences, Poland

4 Department of Histology and Embryology, Wroclaw Medical University, Poland

Abstract

Background. Cell vaults are composed of three proteins (MVP, TEP1, vPARP) and vRNA. They are known to be involved in the development of resistance mechanisms in cancer cells. There is currently a great deal of controversy over how vaults and their components participate in increasing resistance.
Objectives. The absolute quantification of mRNA molecules for MVP, TEP1 and vPARP genes in normal and cancerous cells, and investigation of the quantitative relations between their levels of expression in cells.
Material and Methods. The study involved 18 surgical specimens from patients diagnosed with ovarian cancer. After RNA isolation from the specimens, the expression of genes encoding MVP, TEP1 and vPARP was determined by quantitative reverse-transcription polymerase chain reaction (RT-QPCR) analysis. In some cases gene expression was confirmed by standard immunohistochemistry. The cancer cells in the specimens were identified by standard H+E staining, correlations with the analyzed gene expression levels were calculated using Spearman’s rank correlation technique and the statistical significance was determined.
Results. The absolute quantification of MVP, TEP1 and vPARP mRNA demonstrated a strong correlation between the expression of MVP and TEP1. To some extent, vPARP acts as a housekeeping gene. In the specimen collection used in the current study, the higher level of MVP expression was not associated with a larger number of ovarian cancer cells.
Conclusion. The RT-QPCR analysis revealed that MVP, TEP1 and vPARP genes are widely expressed in normal and cancerous human tissues; however, MVP and TEP1 might associate with each other in order to perform a still-undefined vault function. The high expression of MVP, TEP1 and vPARP also suggests their ubiquitous role in both normal and malignant cells.

Streszczenie

Wprowadzenie. Krypty komórkowe są zbudowane z 3 białek (MVP, TEP1 i vPARP) oraz vRNA. Struktury te uznaje się za czynniki uczestniczące w rozwoju mechanizmów lekooporności komórek nowotworowych. Dotychczas funkcjonowanie krypt wzbudzało wiele kontrowersji, nie poznano, w jaki sposób krypty oraz ich składniki odpowiadają za zwiększenie odporności.
Cel pracy. Bezwzględne określenie liczby cząsteczek mRNA dla genów MVP, TEP1 oraz vPARP w normalnej i nowotworowej komórce oraz określenie relacji między poziomami ich ekspresji w komórce.
Materiał i metody. Zanalizowano 18 próbek chirurgicznych od pacjentów ze zdiagnozowanym rakiem jajnika. Określono ekspresję genów kodujących MVP, TEP1 oraz vPARP za pomocą ilościowej reakcji PCR w czasie rzeczywistym z zastosowaniem w odwrotnej transkrypcji (RT-QPCR) po wcześniejszym wyizolowaniu RNA z próbek poddanych analizie. Potwierdzenie ekspresji genów w pewnych przypadkach uzyskano za pomocą tradycyjnej immunohistochemii. Przeprowadzono ponadto standardowe barwienie H+E w celu oszacowania liczby komórek nowotworowych w próbkach przeznaczonych do izolacji RNA. Korelacje między zanalizowanymi poziomami ekspresji genów obliczono, stosując korelację rang Spearmana oraz wyznaczając istotność statystyczną dla uzyskanych wyników.
Wyniki. Bezwzględna ocena liczby cząsteczek mRNA dla MVP, TEP1 oraz vPARP wykazała silną korelację między ekspresją MVP i TEP1. vPARP wykazuje pewne cechy genu podstawowego (housekeeping gene). W naszym zbiorze próbek wyższa ekspresja MVP nie jest skorelowana z większą liczbą komórek raka jajnika.
Wnioski. Analiza za pomocą RT-QPCR wykazała, że geny MVP, TEP1 oraz vPARP ulegają szerokiej ekspresji w ludzkich tkankach zmienionych nowotworowo oraz kontrolnych, a białka MVP i TEP1 mogą oddziaływać ze sobą i spełniać nieokreśloną jeszcze funkcję krypty komórkowej. Duża ekspresja MVP, TEP1 oraz vPARP wskazuje również na ich powszechną rolę w każdej komórce, normalnej i zmienionej nowotworowo.

Key words

cancer, cell vaults, MVP/LRP, TEP1, vPARP

Słowa kluczowe

nowotwory, krypty komórkowe, MVP/LRP, TEP1, vPARP

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