Piperine Inhibits TGF-β Signaling Pathways and Disrupts EMT-Related Events in Human Lung Adenocarcinoma Cells.

Leonardo Marques Da Fonseca,Leonardo Freire-De-Lima,Lucas Rodrigues Jacques Da Silva,Tatiany Nunes Franklim,Jose Osvaldo Previato,Celio Geraldo Freire De Lima,Marco Edilson Freire De Lima,Julliana De Nazareth Sa-Diniz,Douglas Chaves De Alcântara-Pinto,Jhenifer Santos Dos Reis,Alexandre Morrot,Lucia Mendonça-Previato,Victoria De Sousa Chaves,Marcos André Rodrigues Da Costa Santos,Kelli Monteiro Da Costa

doi:10.3390/medicines7040019

Abstract

Background: Piperine, an amide extracted from the Piper spices, exhibits strong anti-tumor properties. However, its effect on the epithelial–mesenchymal transition (EMT) process has never been investigated. Herein, we evaluate the toxic effect of piperine on lung adenocarcinoma (A549), breast adenocarcinoma (MDA-MB-231) and hepatocellular carcinoma (HepG2) cell lines, as well as its ability to inhibit EMT-related events induced by TGF-β1 treatment. Methods: The cell viability was investigated by MTT assay. Protein expression was evaluated by Western blot. Gene expression was monitored by real-time PCR. Zymography assay was employed to detect metalloproteinase (MMP) activity in conditioned media. Cell motility was assessed by the wound-healing and phagokinetic gold sol assays. Results: The results revealed that piperine was cytotoxic in concentrations over 100 µM, showing IC50 values for HepG2, MDA-MB-231 and A549 cell lines of 214, 238 and 198 µM, respectively. In order to investigate whether piperine would reverse the TGF-β1 induced-EMT, the A549 cell line was pretreated with sublethal concentrations of the natural amide followed by the addition of TGF-β1. Besides disrupting EMT-related events, piperine also inhibited both ERK 1/2 and SMAD 2 phosphorylation. Conclusions: These results suggest that piperine might be further used in therapeutic strategies for metastatic cancer and EMT-related disorders.

Highlights

The discovery of new cancer drugs is a hot topic in cancer research
The cell viability monitored by the MTT assay, showed that 160 and 320 μM of piperine were toxic for all cell lines utilized
Since the acquisition of a spindle-shaped morphology and reduced intercellular adhesion is a fundamental requirement for cell motility, we examined the effect of piperine on the TGF-β1-induced migration of A549 cells

Summary

Introduction

The discovery of new cancer drugs is a hot topic in cancer research. Over the last twenty years, numerous studies have shown that many natural products display chemoprotective properties againstMedicines 2020, 7, 19; doi:10.3390/medicines7040019 www.mdpi.com/journal/medicinesMedicines 2020, 7, x FOR PEER REVIEW MedicinesThe discovery of 16 of new cancer drugs is a hot topic in cancer research. Over the last twenty years, numerous studies have shown that many natural products display chemoprotective properties against. Over the last twenty 2years, numerous studies have shown that many natural products display chemoprotective properties against different types of cancers [1,2]. We evaluate the toxic effect of piperine on lung adenocarcinoma (A549), breast adenocarcinoma (MDA-MB-231) and hepatocellular carcinoma (HepG2) cell lines, as well as its ability to inhibit EMT-related events induced by TGF-β1 treatment.

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Discussion

Conclusion