Par-4 dependent modulation of cellular [beta]-catenin by medicinal plant natural product derivative 3-azido Withaferin A

• Published in: Molecular carcinogenesis 2016-05, Vol.55 (5), p.864
• Main Authors: Amin, Hina, Nayak, Debasis, ur Rasool, Reyaz, Chakraborty, Souneek, Kumar, Anmol, Yousuf, Khalid, Sharma, Parduman Raj, Ahmed, Zabeer, Sharma, Neelam, Magotra, Asmita, Mukherjee, Debaraj, Kumar, Lekha Dinesh, Goswami, Anindya
• Format: Article
• Language: English
• Subjects: AKT protein; Apoptosis; Bcl protein; Bcl-2 protein; Breast cancer; c-Myc protein; Cellular biology; Cyclin D1; E-cadherin; Intracellular; Knock; Localization; Medicinal plants; Metastases; Metastasis; Myc protein; Natural products; Prostate; Prostate apoptosis response 4 protein; Prostate cancer; Proteins; Ras protein; siRNA; Tissues; Transcription; Transfection; β-Catenin
• ISSN: 0899-1987; 1098-2744
• DOI: 10.1002/mc.22328

Here, we provide evidences that natural product derivative 3-azido Withaferin A (3-AWA) abrogated EMT and invasion by modulating [beta]-catenin localization and its transcriptional activity in the prostate as well as in breast cancer cells. This study, for the first time, reveals 3-AWA treatment consistently sequestered nuclear [beta]-catenin and augmented its cytoplasmic pool as evidenced by reducing [beta]-catenin transcriptional activity in these cells. Moreover, 3-AWA treatment triggered robust induction of pro-apoptotic intracellular Par-4, attenuated Akt activity and rescued Phospho-GSK3[beta] (by Akt) to promote [beta]-catenin destabilization. Further, our in vitro studies demonstrate that 3-AWA treatment amplified E-cadherin expression along with sharp downregulation of c-Myc and cyclin D1 proteins. Strikingly, endogenous Par-4 knock down by siRNA underscored 3-AWA mediated inhibition of nuclear [beta]-catenin was Par-4 dependent and suppression of Par-4 activity, either by Bcl-2 or by Ras transfection, restored the nuclear [beta]-catenin level suggesting Par-4 mediated [beta]-catenin regulation was not promiscuous. In vivo results further demonstrated that 3-AWA was effective inhibitor of tumor growth and immunohistochemical studies indicated that increased expression of total [beta]-catenin and decreased expression of phospho-[beta]-catenin and Par-4 in breast cancer tissues as compared to normal breast tissue suggesting Par-4 and [beta]-catenin proteins are mutually regulated and inversely co-related in normal as well as cancer condition. Thus, strategic regulation of intracellular Par-4 by 3-AWA in diverse cancers could be an effective tool to control cancer cell metastasis. Conclusively, this report puts forward a novel approach of controlling deregulated [beta]-catenin signaling by 3-AWA induced Par-4 protein. © 2015 Wiley Periodicals, Inc.