Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2006, vol. 15, nr 6, November-December, p. 989–994

Publication type: original article

Language: English

Influence of N,N−Diethyl−2−[4−(Phenylmethyl) Phenoxy]Ethanamine (DPPE) on the Proliferation of Selected Cell Lines in vitro

Wpływ N,N−dietylo−2−[4−(fenylometylo)fenoksy]etanaminy (DPPE) na proliferację wybranych linii komórkowych in vitro

Anna Merwid−Ląd1,, Małgorzata Trocha1,, Elżbieta Gębarowska2,, Maciej Zabel2,, Adam Szeląg1,

1 Department of Pharmacology, Silesian Piasts University of Medicine in Wrocław, Poland

2 Department of Histology and Embryology, Silesian Piasts University of Medicine in Wrocław, Poland

Abstract

Background. Little is known of the effects of N,N−diethyl−2−[4−(phenylmethyl)phenoxy]ethanamine (DPPE) on cell proliferation. Some studies indicate that DPPE inhibits cell proliferation and differentiation, especially under in vitro conditions, but the in vivo effect is not so clear. It stimulated the growth of chemically induced breast cancer in rat, for example, but inhibited the proliferation of human breast cancer cells in vitro.
Objectives. The aim of this study was to evaluate the effect of DPPE on the proliferation of neoplastic and normal cell lines in vitro.
Material and Methods. The study was carried out on four cell lines: human melanoma (BM), human breast cancer (MCF−7), human gastric cancer (EPG−85−257), and mouse normal fibroblasts (3T3−Balb). They were incubated under standard conditions in medium containing 10% fetal calf serum and a solution of L−glutamine and antibiotics. DPPE was used at concentrations of 400, 40, 4, and 0.4 μg/ml. A cell culture of 2 × 104 cells/ml density was established and 72 hours later the cells were counted, thus providing a preliminary control value. After changing the medium and supplementation with DPPE, the cells were incubated for another 72 hours and counted again. The final control (in the case where DPPE was not added) and final results (in the cases where DPPE was added) were established. The test value and test index were calculated from special formulae.
Results. Human melanoma and mice normal fibroblasts were very sensitive to DPPE in the two higher concentrations (400 and 40 μg/ml); their proliferation was strongly inhibited by DPPE. Human breast cancer and human gastric cancer were slightly sensitive to DPPE in the two higher concentrations; their proliferation was slightly inhibited by DPPE. At a lower concentration (4 μg/ml), only the human melanoma cells were slightly sensitive to DPPE; in this case, the proliferation of human melanoma cells was also inhibited by DPPE, but not as strongly as in the case of the two higher concentrations. Other cell lines were not sensitive to DPPE in the two lower concentrations (4 and 0.4 μg/ml) and proliferation was not inhibited.
Conclusion. Monotherapy of cancer with DPPE is rather not possible due to, among other reasons, its toxicity to normal cells. However, adjuvant therapy with DPPE cannot be excluded. Further studies are necessary.

Streszczenie

Wprowadzenie. Niewiele wiadomo o wpływie N,N−dietylo−2−[4−(fenylometylo)fenoksy]etanaminy (DPPE) na proliferację komórek. Niektóre badania wskazują, że DPPE hamuje proliferację i różnicowanie się komórek, zwłaszcza in vitro, działanie DPPE in vivo nie jest jednak tak jednoznaczne. DPPE pobudza np. wzrost wyindukowanego chemicznie raka sutka u szczurów, chociaż hamuje proliferację komórek raka sutka in vitro.
Cel pracy. Określenie wpływu DPPE na proliferację wybranych linii komórek prawidłowych i nowotworowych w warunkach hodowli in vitro.
Materiał i metody. Badanie zostało przeprowadzone na czterech liniach komórkowych: ludzkiego czerniaka (BM), ludzkiego raka sutka (MCF−7), ludzkiego raka żołądka (EPG−85−257) oraz mysich prawidłowych fibroblastach (3T3−Balb). Wszystkie linie komórkowe były inkubowane w standardowych warunkach, podłoże hodowlane zawierało dodatek 10% płodowej surowicy cielęcej oraz roztwór L−glutaminy i antybiotyków. W doświadczeniu zastosowano następujące stężenia DPPE: 400, 40, 4 i 0,4 μg/ml. Komórki były liczone po 72 godz. od założenia hodowli o gęstości 2 × 104 kom/ml, co było kontrolą wstępną. Po wymianie medium hodowlanego i dodaniu badanego związku w odpowiednich stężeniach komórki były inkubowane przez kolejne 72 godz., a następnie liczone, co było kontrolą ostateczną (gdy nie dodano DPPE) lub wynikiem ostatecznym (gdy dodano DPPE do hodowli). Wartość testu oraz indeks testu wyliczono na podstawie odpowiednich wzorów.
Wyniki. Ludzki czerniak i rak sutka były wrażliwe na działanie DPPE zastosowane w dwóch większych stężeniach (400 i 40 μg/ml). Proliferacja komórek tych dwóch linii została w znacznym stopniu zahamowana przez DPPE. Ludzki rak sutka oraz rak żołądka były mało wrażliwe na działanie DPPE w tych samych stężeniach. Proliferacja komórek tych dwóch linii była hamowana słabiej niż w przypadku poprzednich dwóch linii komórkowych. Gdy DPPE zastosowano w mniejszym stężeniu (4 μg/ml), tylko linia ludzkiego czerniaka była mało wrażliwa. Proliferacja komórek czerniaka ludzkiego była hamowana słabiej niż w przypadku większych stężeń DPPE. Pozostałe linie komórkowe nie były wrażliwe na badany związek zastosowany w dwóch mniejszych stężeniach (4 oraz 0,4 μg/ml), ich proliferacja nie była hamowana przez badany związek.
Wnioski. Terapia chorób nowotworowych samym DPPE wydaje się raczej mało prawdopodobna, chociażby z powodu jego toksyczności wobec komórek prawidłowych. Nie można jednak wykluczyć zastosowania tego związku w terapii wspomagającej klasyczną chemioterapię.

Key words

DPPE, neoplastic cells, proliferation, in vitro studies

Słowa kluczowe

DPPE, komórki nowotworowe, proliferacja, badania in vitro

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