Assessment of Growth Inhibitory, Cell Cycle Arrest, and Apoptosis Inducing Potential of RAS-MAPK Inhibitors in Gemcitabine-Sensitive Pancreatic Cancer Cells

Abstract

Event Abstract Back to Event Assessment of growth inhibitory, cell cycle arrest and apoptosis inducing potential of RAS-MAPK inhibitors in gemcitabine-sensitive pancreatic cancer cells Nityaa Selvarajoo1 and Johnson Stanslas1* 1 Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia Background Pancreatic cancer is an aggressive disease. Although its occurrence rate remains low in Malaysia, it is the third leading cause of cancer-related death in the United States of America because of limited treatment options. Gemcitabine is a standard chemotherapy for pancreatic cancer. Most pancreatic cancers harbour K-Ras mutations that lead to constitutive activation of mitogen-activated protein kinase (MAPK) signalling pathway. The aim of this study was to determine the cell cycle and apoptosis inducing potential of RAS-MAPK inhibitors such as B-Raf kinase inhibitor (vemurafenib), a Mek1/2 kinase inhibitor (selumetinib) and Erk1/2 kinase inhibitor (ulixertinib) in gemcitabine-sensitive pancreatic cancer cell lines. Methods 3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used in assessing in vitro growth inhibition of RAS-MAPK inhibitors against pancreatic cancer cell lines (Panc-1 - mutated KRAS, wild-type BRAF, MEK, and ERK; Colo-357 - mutated KRAS and BRAF, wild-type MEK and ERK; Capan-2 - mutated KRAS, wild-type BRAF, MEK, and ERK); Miapaca-2 - mutated KRAS, wild-type BRAF, MEK, and ERK); BxPC-3- wild-type KRAS, BRAF, MEK and ERK). Cells were treated with 0.1-100 µM of RAS-MAPK inhibitors alone or with gemcitabine and the 50% cell viability- inhibition values were obtained from dose-response curves at 96 hours time-point. Subsequently, flow cytometry was utilised to analyse cell cycle arrest, while Muse® Annexin V & Dead Cell Kit (Merck) was performed to determine apoptosis. Results Ulixertinib exhibited strongest inhibitory effect on Capan-2 with an IC50 value of 0.64 µM, while it showed moderate activity (3.2 µM – 7.25 µM) in other pancreatic cancer cell lines. Vemurafenib and selumetinib were less active against all cell lines. Ulixertinib at 1 µM induced G1 cell cycle arrest in Capan-2. In apoptosis assay, treatment with 1 µM induced 29% and 20.3% of early and late apoptotic cells population, respectively. Conclusion The results suggested that Ulixertinib inhibited growth of Capan-2 cells by inducing G1 arrest and apoptosis. It is presented itself as the most active RAS-MAPK inhibitor against the molecularly selected pancreatic cancer cell lines. Keywords: RAS-MAPK inhibitors, gemcitabine, Apoptosis, Pancreatic Cancer, Cell Cycle Conference: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”, Putrajaya, Malaysia, 3 Dec - 5 Feb, 2019. Presentation Type: Poster Presentation Topic: Cancer Citation: Selvarajoo N and Stanslas J (2019). Assessment of growth inhibitory, cell cycle arrest and apoptosis inducing potential of RAS-MAPK inhibitors in gemcitabine-sensitive pancreatic cancer cells. Front. Pharmacol. Conference Abstract: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”. doi: 10.3389/conf.fphar.2018.63.00145 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 26 Sep 2018; Published Online: 17 Jan 2019. * Correspondence: Prof. Johnson Stanslas, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia, rcxjs@upm.edu.my Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Nityaa Selvarajoo Johnson Stanslas Google Nityaa Selvarajoo Johnson Stanslas Google Scholar Nityaa Selvarajoo Johnson Stanslas PubMed Nityaa Selvarajoo Johnson Stanslas Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.