Curcumin rescue p53Y220C in BxPC-3 pancreatic adenocarcinomas cell line: Evidence-based on computational, biophysical, and in vivo studies

• Published in: Biochimica et biophysica acta. General subjects Vol. 1865; no. 2; p. 129807
• Main Authors: Malhotra, Lakshay, Goyal, Harsh K.V., Jhuria, Sunita, Dev, Kapil, Kumar, Saroj, Kumar, Manoj, Kaur, Punit, Ethayathulla, Abdul S.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2021
• Subjects: adenocarcinoma; Adenocarcinoma - drug therapy; Adenocarcinoma - genetics; Annexin V/ PI; Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis - drug effects; caspase-3; CD spectroscopy; cell cycle checkpoints; Cell Line, Tumor; cell lines; curcumin; Curcumin - pharmacology; DNA; DNA-binding domains; Docking; fluorescence; Fluorescence spectroscopy; FTIR spectroscopy; Humans; infrared spectroscopy; Molecular Docking Simulation; mutants; mutation; Pancreatic Neoplasms - drug therapy; Pancreatic Neoplasms - genetics; phosphotransferases (kinases); Point Mutation - drug effects; therapeutics; Tumor Suppressor Protein p53 - genetics; tumor suppressor proteins; Fluorescence spectroscopy; Annexin V/ PI; p53wt; IPTG; Docking; p53Y220C; CD spectroscopy; FTIR spectroscopy; Apoptosis
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2020.129807

The p53, tumor suppressor protein is inactivated upon mutation in the DNA-binding domain and the non-functional protein leads to cancers. The p53Y220C is one of the most frequently observed mutations in p53 with a scope of rescuing the protein function using small molecules. Using computational modeling, biophysical, and experimental cell-based studies we tried to understand the molecular basis of Curcumin as a potential small molecule to stabilize p53Y220C mutant and restore its function. The pancreatic adenocarcinomas BxPC-3 p53Y220C mutant cell line was used for cell-based assays to determine the therapeutic potential of Curcumin to restore mutant p53 to function like wild type. Our results showed that the Curcumin binds p53Y220C with Kd = 3.169 ± 0.257 μM and it increases the DNA binding affinity of the mutant by 4-fold with Kd = 851.29 ± 186.27 nM. By Fluorescence, CD, and IR spectroscopy, we could characterize the secondary structural changes and stabilization of the p53Y220C DNA binding domain upon Curcumin binding. By caspase-3 and Annexin V assays, we could demonstrate that Curcumin at 3 μM to 8 μM concentration could initiate p53 mediated apoptosis in BxPC-3 cell line. Based on our experimental studies, we propose a mechanism for the activation of ATM/Chk1 kinases pathways for apoptosis and/or G2/M cell cycle arrest in the BxPC-3 cell line mediated by functionally restored p53Y220C. The study indicated that the natural compound Curcumin could rescue mutant p53Y220C in BxPC-3 pancreatic adenocarcinomas cell line to function like wild-type and activate apoptotic pathways. [Display omitted] •Natural compound Curcumin rescue p53Y220C mutant.•Computational, IR, CD studies establish secondary structural changes.•Cell-based MTT, Annexin V and Caspase3 assays establish apoptosis induced by Curcumin.•Curcumin could induce apoptosis in human pancreatic cancer cells.