Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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Advances in Clinical and Experimental Medicine

2010, vol. 19, nr 5, September-October, p. 585–591

Publication type: original article

Language: English

The Expression of Genes Encoding Antioxidant Enzymes in Lung Adenocarcinoma A549 Cells Exposed to Cisplatin Supplemented by Polyunsaturated Fatty Acids

Ekspresja genów kodujących enzymy antyoksydacyjne w suplementowanych wielonienasyconymi kwasami tłuszczowymi komórkach gruczolakoraka płuca A549 eksponowanych na cisplatynę

Alicja Zajdel1,, Adam Wilczok1,, Arkadiusz Gruchlik1,, Piotr Paduszyński1,, Zofia Dzierżewicz1,

1 Department of Biopharmacy, Medical University of Silesia, Katowice, Poland

Abstract

Background. The effectiveness of cisplatin (CPT) in the treatment of tumors is often limited by primary or acquired resitance. Supplementation with polyunsaturated fatty acids, in particular eicosapentaenoic acid (EPA, 20 : 5, n-3) and docosahexaenoic acid (DHA, 22 : 6, n-3), can enhance drug sensitivity of tumor cells to anticancer therapy or reverse cell resistance. Antioxidant enzymes, highly expressed in lung tumors, are among the determinant factors in the sensitivity of lung tumors to chemotherapy.
Objectives. Investigation of the effect of EPA and DHA at different concentrations on gene expression of selected enzymes controlling cell redox status, such as superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2), catalase (CAT), phospholipid hydroperoxide glutathione peroxidase (GPx-4), and glutathione S-transferase pi (GST-pi) in human lung adenocarcinoma cells (A549) exposed to CPT.
Material and Methods. Viability of A549 cells treated with CPT and EPA or DHA was measured using the XTT tetrazolium salt based assay. Expression of genes encoding the antioxidant enzymes was determined by quantitative reverse-transcription polymerase chain reaction (QRT-PCR) analysis after RNA isolation from A549 cells.
Results. EPA and DHA added to the culture medium, increased the antitumor activity of CPT in A549 cells in a concentration dependent manner. The SOD2 and GST-pi expression showed marked increase after CPT treatment, while supplementation with EPA and DHA down-regulated their expression in the CPT treated A549 cells. The observed changes in mRNA levels of CAT were not statistically significant.
Conclusion. The reduction of antioxidant potential in cancer cells may sensitize these cells to anticancer therapy. PUFAs supplementation during CPT-based anticancer therapy may enhance effectiveness of the treatment.

Streszczenie

Wprowadzenie. Pierwotna lub nabyta oporność często ogranicza zastosowanie cisplatyny (CPT) w leczeniu nowotworów. Suplementacja wielonienasyconymi kwasami tłuszczowymi (w.n.k.t.), w szczególności eikozapentaenowym (EPA, 20 : 5, n-3) i dokozaheksaenowym (DHA, 22 : 6, n-3), może zwiększać wrażliwość komórek nowotworowych na leki albo znosić ich oporność. Zwiększona ekspresja enzymów antyoksydacyjnych w nowotworach płuca jest czynnikiem determinującym ich wrażliwość na chemioterapię.
Cel pracy. Zbadanie wpływu EPA i DHA, w różnych stężeniach, na ekspresję genów kodujących wybrane enzymy kontrolujące potencjał redox: dysmutazę ponatlenkową 1 (SOD1), dysmutazę ponatlenkową 2 (SOD2), katalazę (CAT), peroksydazę glutationowa wodoronadtlenków fosfolipidów (GPx-4) i S-transferazę glutationową pi (GST-pi) w ludzkich komórkach gruczolakoraka płuca (A549) eksponowanych na CPT.
Materiał i metody. Przeżywalność komórek A549 eksponowanych na CPT i EPA lub DHA była mierzona za pomocą testu wykorzystującego sól tetrazoliową XTT. Ekspresja genów kodujących ww. enzymy antyoksydacyjne w ekstraktach RNA z komórek A549 została oznaczona metodą ilościowej reakcji łańcuchowej polimerazy poprzedzonej odwrotną transkrypcją (QRT-PCR).
Wyniki. EPA i DHA dodane do medium hodowlanego zwiększały, w sposób zależny od stężenia, przeciwnowotworową aktywność CPT. Suplementacja w.n.k.t. komórek A549 eksponowanych na CPT zmniejszała ekspresję SOD2 i GST-pi, która po ekspozycji tylko na CPT była znacznie podwyższona.
Wnioski. Ograniczenie potencjału antyoksydacyjnego w komórkach nowotworowych może zwiększać ich wrażliwość na terapię przeciwnowotworową. Suplementacja w.n.k.t. podczas terapii przeciwnowotworowej z udziałem CPT może poprawić skuteczność leczenia.

Key words

antioxidant enzymes, PUFAs, cisplatin, A549 cells

Słowa kluczowe

enzymy antyoksydacyjne, w.n.k.t., cisplatyna, komórki A549

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