Goniothalamin Induces Necroptosis and Anoikis in Human Invasive Breast Cancer MDA-MB-231 Cells.

Patompong Khaw-On,Ratana Banjerdpongchai,Wilart Pompimon

doi:10.3390/ijms20163953

Patompong Khaw-On, Ratana Banjerdpongchai + Show 1 more

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https://doi.org/10.3390/ijms20163953

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Abstract

Goniothalamin (GTN) is toxic to several types of cancer cells in vitro. However, its effects on non-apoptotic cell death induction of human cancer cells have been poorly documented. Here, an investigation of the anti-cancer activity of GTN and the molecular signaling pathways of non-apoptotic cell death in the invasive human breast cancer MDA-MB-231 cell line were undertaken. Apoptotic cell death was suppressed by using a pan-caspase inhibitor (Benzyloxycarbonyl-Val-Ala-Asp-[O-methyl]-fluoromethylketone), z-VAD-fmk) as a model to study whether GTN induced caspase-independent cell death. In the anoikis study, MDA-MB-231 cells were cultured on poly-(2-hydroxyethyl methacrylate)- or poly-HEMA- coated plates to mimic anoikis-resistance growth and determine whether GTN induced cell death and the mechanisms involved. GTN and z-VAD-fmk induced human breast cancer MDA-MB-231 cells to undergo necroptosis via endoplasmic reticulum (ER) and oxidative stresses, with increased expressions of necroptotic genes such as rip1, rip3, and mlkl. GTN induced MDA-MB-231 cells to undergo anoikis via reversed epithelial-mesenchymal transition (EMT) protein expressions, inhibited the EGFR/FAK/Src survival signaling pathway, and decreased matrix metalloproteinase secretion.

Highlights

Goniothalamin (GTN) is a phytosteryl-lactone found in the genus Goniothalamus and has been widely used as a folk medicine in Southeast Asia
Death receptors including tumor necrosis factor receptor (TNFR), Fas, and tumor necrosis factor-relating apoptosis-induced ligand-receptor (TRAIL-R) activate necroptosis by recruiting necrosome formation consisting of receptor-interacting serine/threonine protein kinases 1 (RIP1) and RIP3 [8,9]
The inhibitory concentration at 50 percent (IC50) was 33.82 μM compared to 44.65 μM without z-VAD-fmk, but GTN was less cytotoxic to MCF-10A, which is a non-tumorigenic human breast epithelial cell line with an IC50 of >80 μM

Summary

Introduction

Goniothalamin (GTN) is a phytosteryl-lactone found in the genus Goniothalamus and has been widely used as a folk medicine in Southeast Asia. GTN induces necrosis in MCF-7 cells [7] through mechanisms that are still not completely understood. Necroptosis, a programmed form of necrosis, is regulated through caspase-independent cell death mechanisms. Mixed-lineage kinase domain-like (MLKL) protein, a substrate of necrosome, translocates to plasma membrane forming channels, which leads to the influx of Ca2+ ions and subsequently necroptotic cell death [10]. Alkylating DNA-damage agents trigger caspase-independent necroptosis, which involves the serial activation of various proteins and enzymes including PARP-1, calpains, Bcl-2 associated X protein (Bax), and apoptosis-inducing factor (AIF) as crucial regulators of a rapid increase in inner mitochondrial membrane permeability and caspase-independent necroptosis [11,12]

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