Gene expression profiling identifies novel key players involved in the cytotoxic effect of Artesunate on pancreatic cancer cells

Abstract

Pancreatic cancer is one of the most aggressive human malignancies, with an extremely poor prognosis. The paucity of curative therapies has translated into an overall 5-year survival rate of less than 5%, underscoring a desperate need for new therapeutic options.Artesunate (ART), clinically used as anti-malarial agent, has recently revealed remarkable anti-tumor activity. However, the mechanisms underlying those activities in pancreatic cancer are not yet known. Here we evaluated the antitumor activity of Artesunate and the possible underlying mechanisms in pancreatic cancer. MiaPaCa-2 (poorly differentiated) and BxPC-3 (moderately differentiated) pancreatic cancer cell lines were treated with Artesunate and the effect was monitored by a tetrazolium-based assay (MTT) for evaluating cell viability and by flow cytometry and caspase 3/7 activation for apoptosis evaluation. In addition cDNA arrays were used to identify differentially expressed genes. The microarray data were then validated by RT-PCR and western blotting. Moreover, pathways associated with these expression changes were identified using the Ingenuity Pathway Analysis. The expression analysis identified a common set of genes that were regulated by Artesunate in pancreatic cancer. Our results provide the first in vitro evidence for the therapeutic utility of Artesunate in pancreatic cancer. Moreover, we identified Artesunate as a novel topoisomerase IIα inhibitor that inhibits pancreatic cancer growth through modulation of multiple signaling pathways. The present analysis is a starting point for the generation of hypotheses on candidate genes and for a more detailed dissection of the functional role of individual genes for the activity of Artesunate in tumor cells.

Genotoxic evaluation of the antimalarial drugs artemisinin and artesunate in human HepG2 cells and effects on CASP3 and SOD1 gene expressions

“…Our findings also agree with authors who assessed the genotoxic effects of other sesquiterpenes (Burin et al, 2001;Canalle et al, 2001;Al-Zubairi et al, 2010). In addition to causing DNA damage, the cytotoxicity of these substances, especially artemisinin, is due in part to the decreased expression of topoisomerase 2 (Youns et al, 2009), which is essential for cell proliferation and is expressed in dividing cells (Toyoda et al, 2008).…”

“…Hou et al (2008) reported that artemisinin and artesunate exerted the greatest cytotoxicity on HepG2, inhibiting cell proliferation and inducing apoptosis by activating the CASP3 gene. Other authors also observed similar effects for both of these compounds (Li et al, 2008;Youns et al, 2009;Du et al, 2010;Zhang et al, 2010). We analyzed expression of the CASP3 gene in HepG2 cells after exposure to artemisinin and artesunate because it is the final common pathway of almost all apoptotic pathways.…”

“…Some in vitro studies using cancer cell lines suggested that artemisinin and some of its derivatives have cytotoxic effects, alter the cell cycle, and induce apoptosis (Hou et al, 2008;Li et al, 2008;Youns et al, 2009;Zhang et al, 2010). These compounds may present cytotoxic activity in mammal cell proliferation (Efferth et al, 2001;Disbrow et al, 2005), and it has been suggested that the toxicity is related to high intracellular concentrations of iron (Smith et al, 1997;Singh and Lai, 2001).…”

Synergism of Cytotoxicity Effects of Triptolide and Artesunate Combination Treatment in Pancreatic Cancer Cell Lines

“…Recently, tumor therapy by traditional Chinese herb is becoming more and more attractive (He et al, 2008;Youns et al, 2009;Chen et al, 2010). We therefore attempted to examine the effects of triptolide and artesunate on pancreatic cancer.…”

A Novel Peptide from <i>Vespa ducalis</i> Induces Apoptosis in Osteosarcoma Cells by Activating the p38 MAPK and JNK Signaling Pathways

“…Cell Cycle and Apoptosis of OS Cells by Flow Cytometer Assays Cells treated as described were fixed with 95% ethanol, washed with cold PBS and incubated with 150 mL of hypotonic fluorochrome solution, as previously described, 20) in the dark at 4°C overnight. The apoptotic cells were stained using a CF488A-Annexin V and Propidium Iodide Apoptosis Assay Kit (Biotium), according to the protocols of the kit.…”