Vol.:(0123456789) 1 3 Molecular Biology Reports https://doi.org/10.1007/s11033-018-4294-5 ORIGINAL ARTICLE A natural favonoid lawsonaringenin induces cell cycle arrest and apoptosis in HT-29 colorectal cancer cells by targeting multiple signalling pathways Areeba Anwar 1  · Nizam Uddin 2  · Bina Shaheen Siddiqui 2  · Rafat Ali Siddiqui 3  · Sabira Begum 2  · Muhammad Iqbal Choudhary 1,2,4 Received: 11 May 2018 / Accepted: 1 August 2018 © Springer Nature B.V. 2018 Abstract Colorectal cancer is the third most common malignancy in the world having a high mortality rate. Flavonoids possess many biological activities including anti-cancer activity. lawsonaringenin (LSG) is a favonoid isolated from leaves of Lawsonia alba Lam. The objective of this study was to demonstrate the anti-cancer potential of LSG in colorectal cancer for the frst time. The HT-29 cells were treated with LSG or 5-fuoruracil, as a positive control, to determine its efect on cell cytotoxic- ity by a MTT cell proliferation assay, and cell cycle progression and apoptosis using fowcytometry. We also determined the mechanisms underlying LSG-mediated growth inhibition of HT-29 cells by by investigating the expression of key oncogenes and apoptosis genes using q-RT PCR and immunocytochemical analysis. The cell cytotoxicity data showed that the IC 50 value of LSG was signifcantly less than the IC 50 value of 5-FU (50 µM). The anti-proliferative efect of LSG was mediated by arresting cells in the S phase of the cell cycle which then led to the induction of apoptosis the q-RT PCR and immunocyto- chemical analysis showed that LSG reduced the expression of ß-catenin (non-phosphorylated) and its downstream signalling target c-Myc, whereas it increased the phosphorylation of ß-catenin. Furthermore, LSG also downregulated the expression of oncogene K-Ras and anti-apoptotic proteins, Bcl-2, and Bcl-xL. In conclusion, our data demonstrates that LSG exerted its anti-tumor activity by arresting the cell cycle in S phase, and by downregulating the expression of oncogenes including ß-catenin, c-Myc, K-Ras and anti-apoptosis proteins Bcl-2 and Bcl-xL. This study suggests a potential use of natural favo- noid, lawsonaringenin, to attenuate colorectal cancer growth; however, further pre-clinical/clinical studies are required to establish its role as a therapeutic agent. Keywords Colorectal cancer · HT-29 cells · Lawsonia alba Lam. · Apoptosis · ß-Catenin signalling · Flavonoid Abbreviations ß-Cat-NP Beta catenin (non-phosphorylated) ß-Cat-P Beta catenin (phosphorylated) 5-FU 5-Fluorouracil Bcl-2 B-cell lymphoma 2 protein Bcl-Xl B-cell lymphoma extra-large protein Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-018-4294-5) contains supplementary material, which is available to authorized users. * Areeba Anwar areeba_anwar@ymail.com * Muhammad Iqbal Choudhary iqbal.choudhary@iccs.edu 1 Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan 2 H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan 3 Nutrition Science and Food Chemistry Laboratory, Agriculture Research Station, Virginia State University, Petersburg, VA 23806, USA 4 Department of Biochemistry, Faculty of Sciences, King Abdulaziz University, Jeddah 21412, Saudi Arabia