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Planta Med 2014; 80(06): 473-481
DOI: 10.1055/s-0034-1368301
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

The Anti-Promyelocytic Leukemia Mode of Action of Two Endophytic Secondary Metabolites Unveiled by a Proteomic Approach

Margareth B. C. Gallo
1   Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA, USA
2   Department of Pharmaceutical Sciences, Faculdade de Ciências Farmacêuticas de Ribeirão Preto – USP, Ribeirão Preto, SP, Brazil
3   Mass Spectrometry Core Facility, Centro de Pesquisas Gonçalo Moniz (CPqGM) – FIOCRUZ, Salvador, BA, Brazil
,
Miranda J. S. Falso
4   Department of Pharmacology & Chemical Biology, University of Pittsburgh, Pittsburgh, PA, USA
,
Fernanda Balem
5   Department of Structural Biology, University of Pittsburgh, Pittsburgh, PA, USA
,
Diego Menezes
3   Mass Spectrometry Core Facility, Centro de Pesquisas Gonçalo Moniz (CPqGM) – FIOCRUZ, Salvador, BA, Brazil
6   Núcleo de Biotecnologia e Bioprospecção, Escola Bahiana de Medicina e Saúde Pública, Salvador, BA, Brazil
,
Núbia Rocha
6   Núcleo de Biotecnologia e Bioprospecção, Escola Bahiana de Medicina e Saúde Pública, Salvador, BA, Brazil
,
Raghavan Balachandran
1   Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA, USA
,
Timothy S. Sturgeon
7   Center for Vaccine Research, University of Pittsburgh, Pittsburgh, PA, USA
,
Mônica T. Pupo
2   Department of Pharmaceutical Sciences, Faculdade de Ciências Farmacêuticas de Ribeirão Preto – USP, Ribeirão Preto, SP, Brazil
,
Billy W. Day
1   Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA, USA
› Author Affiliations
Further Information

Publication History

received 20 December 2013
revised 27 January 2014

accepted 22 February 2014

Publication Date:
07 April 2014 (online)

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Abstract

As a result of a program to find antitumor compounds of endophytes from medicinal Asteraceae, the steroid (22E,24R)-8,14-epoxyergosta-4,22-diene-3,6-dione (a) and the diterpene aphidicolin (b) were isolated from the filamentous fungi Papulaspora immersa and Nigrospora sphaerica, respectively, and exhibited strong cytotoxicity against HL-60 cells. A proteomic approach was used in an attempt to identify the drugsʼ molecular targets and their respective antiproliferative mode of action. Results suggested that the (a) growth inhibition effect occurs by G2/M cell cycle arrest via reduction of tubulin alpha and beta isomers and 14–3–3 protein gamma expression, followed by a decrease of apoptotic and inflammatory proteins, culminating in mitochondrial oxidative damage that triggered autophagy-associated cell death. Moreover, the decrease observed in the expression levels of several types of histones indicated that (a) might be disarming oncogenic pathways via direct modulation of the epigenetic machinery. Effects on cell cycle progression and induction of apoptosis caused by (b) were confirmed. In addition, protein expression profiles also revealed that aphidicolin is able to influence microtubule dynamics, modulate proteasome activator complex expression, and control the inflammatory cascade through overexpression of thymosin beta 4, RhoGDI2, and 14–3–3 proteins. Transmission electron micrographs of (b)-treated cells unveiled dose-dependent morphological characteristics of autophagy- or oncosis-like cell death.

Key words

Chaetomiaceae - Papulaspora immersa - Trichosphaeriaceae - Nigrospora sphaerica - aphidicolin - epoxyergostane-type steroid
 

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