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

2010, vol. 19, nr 6, November-December, p. 679–684

Publication type: original article

Language: English

The Influence of Vitamin D Derivatives on the Expression of Apoptotic Genes in Nasal Polyp Fibroblasts

Ocena wpływu pochodnych witaminy D na ekspresję genów BCL-2 i BAX w fibroblastach polipów nosa

Marcin Frączek1,, Beata Rostkowska-Nadolska1,, Elektra Sliupkas-Dyrda2,, Dariusz Kuśmierz2,, Jakub Pniak2,, Małgorzata Latocha2,

1 Department of Otolaryngology, Wroclaw Medical University, Wrocław, Poland

2 Department of Cell Biology, Medical University of Silesia, Sosnowiec, Poland

Abstract

Background. Vitamin D (VD) modulates the transcription of genes involved in a variety of biological processes, such as phosphocalcic metabolism, proliferation, differentiation, angiogenesis, cell division and cell death. The pro-apoptotic and antiproliferative effects of VD derivates, although still not fully clear, could potentially be harnessed therapeutically.
Objectives. The aim of the paper was to analyze the influence of VD derivatives on the expression of apoptosis associated genes in fibroblasts derived from nasal polyps (NP).
Material and Methods. During routine endonasal surgery 16 NP samples were collected from 5 women and 11 men suffering from chronic rhinosinusitis with NP. The expression of BCL-2 and BAX was detected using RT-PCR and, at the protein level, via ELISA in fibroblast cultures treated with either calcitriol (1,25-(OH)2D3) or tacalcitol (1,24-(OH)2D3) at concentrations of 10–6M, 10–5M and 10–4M each. The comparison group consisted of NP fibroblasts not treated with VD derivatives.
Results. Both tacalcitol and calcitriol significantly decreased BCL-2 and BAX expression at a concentration of 10–4M (p < 0.05). An insignificant decrease in BCL-2/BAX ratios was noted after treatment with both of the examined VD derivatives, regardless of the drug concentration. The highest pro-apoptotic ratio was noted with tacalcitol at a concentration of 10–5M. The decrease in the ratio was more a result of inhibited BCL-2 expression than of enhanced BAX expression. Concentration-dependent action was not seen for either of the examined substances.
Conclusion. The results imply that the pro-apoptotic properties of VD derivatives may engage the BCL-2 family pathway only to a limited extent. The slightly better efficiency of tacalcitol in reducing the BCL-2/BAX mRNA ratio, along with its lower calcemic effects, makes tacalcitol preferable to calcitriol.

Streszczenie

Wprowadzenie. Witamina D, oprócz wpływu na gospodarkę wapniowo-fosforanową, moduluje również ekspresję genów uczestniczących w wielu ważnych procesach komórkowych, takich jak: proliferacja, podział, różnicowanie i śmierć komórki. Nie w pełni poznane proapoptotyczne i antyproliferacyjne właściwości witaminy D mają duży potencjał terapeutyczny.
Cel pracy. Ocena wpływu pochodnych witaminy D (kalcytriolu i takalcytolu) na ekspresję genow związanych z procesem apoptozy w hodowli fibroblastów pochodzących z polipów nosa.
Materiał i metody. Materiał do badania został uzyskany podczas operacji endoskopowej od 16 pacjentów (5 kobiet i 11 mężczyzn) chorujących na przewlekłe zapalenie zatok przynosowych. Ekspresję genów BCL-2, BAX oceniono metodą RT-PCR oraz na poziomie białka metodą ELISA w hodowli fibroblastów z dodatkiem kalcytriolu lub takalcytolu w stężeniu 10–6M, 10–5M i 10–4M. Grupę porównawczą stanowiły fibroblasty z polipów nosa nietraktowane pochodnymi witaminy D.
Wyniki. Zarówno kalcytriol, jak i takalcytol w stężeniu 10–4M istotnie zmniejszał ekspresję BCL-2 i BAX. Niezależnie od stężenia żadna z substancji nie obniżyła istotnie statystycznie wartości współczynnika BCL-2/BAX. Współczynnik BCL-2/BAX osiągnął najbardziej proapoptotyczną wartość po dodaniu takalcytolu w stężeniu 10–5M. Obniżenie jego wartości było rezultatem raczej zahamowania ekspresji BCL-2 niż zwiększenia ekspresji BAX.
Wnioski. Wyniki sugerują, że proapoptotyczne działanie witaminy D tylko w ograniczonym zakresie opiera się na rodzinie białek BCL-2. Nieznacznie bardziej korzystny wpływ na stosunek ekspresji BCL-2 do BAX oraz słabszy wpływ na gospodarkę wapniową sprawiają, że takalcytol jest lepszy niż kalcytriol.

Key words

calcitriol, tacalcitol, nasal polyps, QRT-PCR, immunohistochemistry, ELISA, apoptosis

Słowa kluczowe

kalcytriol, takalcytol, polipy nosa, QRT-PCR, immunohistochemia, ELISA, apoptoza

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