Abstract 4307: Molecular basis of cell cycle arrest induced by erufosine in metastatic breast cancer cells

Abstract

Abstract Introduction: Breast cancer, a major health care burden with limited curative options for advanced metastatic phase, demands a search for novel therapeutic compounds. Synthetic alkyl-phospholipids (ALPs) comprise a promising class of anti-cancer agents in this context, where 1st and 2nd generation compounds are already in clinical use to treat various malignancies. Erufosine is the latest (3rd) generation of ALPs and has shown promising anti-tumour activities against breast cancer with minimal toxicity towards GI tract, negligible haemolytic activity and a good pharmacological profile in laboratory settings. Previously, we highlighted the significant potential of erufosine to induce arrest in G2/M phase of cell cycle in breast cancer cells. Currently, we decipher the mechanistic basis of these observed effects at molecular levels. Materials & Methods: Metastatic breast cancer cells (MDA-MB-231) were exposed to low (IC25, 12.3µM), medium (IC50, 19.5µM) and high (IC75, 30.2µM) concentrations of erufosine for 48h. Distribution of the cells in different phases of the cell cycle was determined by propidium iodide based labelling of the DNA and flow cytometry analysis. Expressional modulations in 84 cell cycle relevant genes in response to erufosine exposure (IC75, 48h) were studied by using a ready-made panel (Qiagen, Cat#330231) and real-time PCR methodology. Alterations in expressional profile of significantly altered genes (≥2fold) were used to design a signalling pathway with the help of Ingenuity Pathway Analysis at core facility of DKFZ, Heidelberg, Germany. Results: Exposure with erufosine induced a significant arrest in the G2/M phase of cell cycle in MDA-MB-231 cells in a concentration dependant format. Significant alterations (≥2fold) were found in 39% (33/84) of the cell cycle relevant genes incorporated in the ready-made panel. Most of the altered genes (28/33) were down-regulated in response to erufosine exposure. Significantly altered genes belong to various groups like kinases (ATM, AURKA, B), phosphatases (CDC25A, C), tumour suppressors (BRCA1), anti-apoptotic genes ( BCL2, BIRC5), cyclins (CCNA2, CCNB1, CCNB2, CCND2, CCNF, CCNG2), their target kinases (CDKs), activators (CDC6, MK167), inhibitors (CDKN1A, 2A, CDKN3, CKS1B), regulators (CDC20, MKI67), facilitators of the cell cycle (MCM2, 3, 4) and transcription factors (CKS2, E2F1). Conclusion: Erufosine is a significant cytostatic agent and induces a major halt in the G2/M phase of cell cycle in metastatic breast cancer cells. The compound imposes noteworthy alterations in various cell cycle relevant genes to cause these cytostatic effects. Erufosine, provided with further studies in laboratory (in vitro and in vivo) and clinical settings, could be a valuable therapeutic agent for metastatic breast cancer. Citation Format: Asim Pervaiz, Muhammad Shoaib Akhtar, Saqib Mahmood, Osheen Sajjad, Saba Khaliq, Martin R. Berger. Molecular basis of cell cycle arrest induced by erufosine in metastatic breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4307.