PSEUROTIN A FROM Aspergillus fumigatus Fr. AUMC 8002 EXHIBITS ANTICANCER ACTIVITY AGAINST HEPATOCELLULAR CARCINOMA IN VITRO AND IN VIVO

Abstract

This study investigated the in vitro and in vivo anti-hepatocellular carcinoma activity of pseurotin A isolated from the n-butanol extract of Aspergillus fumigatus Fr. AUMC 8002 (nBE-AF).The in vitro anticancer activity of nBE-AF was measured against a human hepatocellular carcinoma cell line (HepG2) using a sulforhodamine-B (SRB) assay. The intraperitoneal median lethal dose (LD50) of nBE-AF was determined in rats. Hepatocellular carcinoma was induced in rats by a single intraperitoneal injection of diethylnitrosamine (DEN) (200 mg/kg b.wt.) followed by subcutaneous injections of carbon tetrachloride (CCl4) (3 ml/kgb.wt.) weekly for 6 weeks. After administration of these carcinogens, 1/10 and 1/20 LD50 doses of nBE-AF were administered intraperitoneally daily. NBE-AF exhibited significant cytotoxic activity against HepG2 cells. Administration of DEN and CCl4 significantly elevated the serum levels of liver function and tumour markers and significantly downregulated tumour necrosis factor-α gene expression. Moreover, DEN and CCl4 decreased immunohistochemical Bax expression and increased Bcl-2 expression in the liver. Co-treatment with nBE-AF mitigated the DEN+CCl4-induced alterations in a dose-dependent manner. Histopathological evaluation of the liver substantiated the above biochemical results. These results confirmed that nBE-AF, via its major isolated secondary metabolite, pseurotin A, exerted an anti-hepatocarcinogenic effect and could be used as a chemopreventive agent for hepatocellular carcinoma.awareness of the role of mycobacteria in infections associated with exposure to aquarium fish. Key words: Aspergillus fumigatus extract; pseurotin A; hepatocellular carcinoma; anticancer; cytotoxic PSEUROTIN A IZ GLIVE Aspergillus fumigatus Fr. AUMC 8002 ZAVIRA RAST KARCINOMA JETER IN VITRO IN IN VIVO Povzetek: V študiji so raziskali in vitro in in vivo protirakasto aktivnost pseurotina A, pridobljenega iz n-butanolnega izvlečka glive Aspergillus fumigatus Fr. AUMC 8002 (nBE-AF), na karcinom jeter. In vitro protirakasta aktivnost nBE-AF je bila spremljana pri humani celični liniji jetrnega karcinoma (HepG2) z uporabo analize sulforhodamin-B (SRB). Pri podganah so intraperitonealno določili povprečni smrtni odmerek (LD ) nBE-AF. Jetrni karcinom je bil pri podganah sprožen z enkratno intraperitonealno injekcijo dietilnitrozamina (DEN) (200 mg/kg telesne teže), čemur so sledile podkožne injekcije ogljikovega tetraklorida (CCl 50 ) (3 ml/ kg) enkrat tedensko 6 tednov. Po dodajanju teh rakotvornih snovi so 1/10 in 1/20 odmerka LD nBE-AF aplicirali vsak dan intraperitonealno. NBE-AF je pokazal pomembno citotoksično aktivnost proti celicam HepG2. Uporaba DEN in CCl4 je značilno povečala serumske vrednosti jetrnih funkcij in tumorskih markerjev ter znatno zmanjšala izražanje gena za faktor tumorske nekroze α (TNF- α). Poleg tega sta DEN in CCl 4 50 zmanjšala imunohistokemično izražanje gena Bax in povečala izražanje gena Bcl-2 v jetrih. Sočasno zdravljenje z nBE-AF je zmanjšalo z DEN + CCl sprožene spremembe v jetrih v povezavi z velikostjo odmerka. Histopatološka ocena jeter je potrdila zgoraj navedene biokemične rezultate. Rezultati so potrdili, da je nBE-AF preko svojega glavnega izoliranega sekundarnega metabolita pseurotina A izkazal protirakasti učinek na jetra in bi se lahko v prihodnosti morda uporabljal kot zdravilo za zdravljenje karcinoma jeter. Ključne besede: izvleček Aspergillus fumigatus; pseurotin A; karcinom jeter; protirakasto; citotoksično