# Modulatory Effects of Cirsimarin on Lung Cancer A549 Cells Migration in 2D and 3D Cultures Involves Transcriptional Regulation of Metalloproteinases

## TL;DR

This study shows that cirsimarin, a flavone, inhibits lung cancer cell migration and viability in 2D and 3D models by affecting genes related to cell growth and matrix remodeling.

## Contribution

The novel contribution is the identification of cirsimarin's antimetastatic potential through transcriptional regulation of metalloproteinases in lung cancer cells.

## Key findings

- Cirsimarin reduced A549 cell viability and migration in 2D and 3D models.
- It induced apoptosis and DNA damage at specific concentrations.
- Downregulation of genes like MMP-2, MMP-9, and TP53 was observed.

## Abstract

Current toxicology and cancer biology investigations have focused on developing alternative models that better recapitulate the in vivo architecture of tissues and organs. The present study evaluated the anticancer effects of the flavone cirsimarin, which presented successful antitumor activity on breast tumor cells. We assessed the impact of flavone on cell viability, proliferation, and migration, as well as on DNA integrity and modulation of related cellular pathways. In the 2D model, cirsimarin reduced cell viability at concentrations ≥ 80 μM after 24 h of treatment (resazurin assay), selectively in A549 cells compared to MRC‐5 non‐tumor cells. Apoptosis was induced at concentrations ≥ 40 μM, and clonogenicity was reduced by approximately 50% only at 160 μM. In the wound healing assay, cirsimarin (1–80 μM) completely inhibited cell migration and induced DNA damage (comet assay). These apoptotic and anti‐migratory effects were associated with the downregulation of key genes involved in cell proliferation, death, and extracellular matrix remodeling, including TNF‐α (0.32‐fold), TP53 (0.17‐fold), MMP‐2 (0.18‐fold), MMP‐9 (0.43‐fold), and MMP‐11 (0.04‐fold), as revealed by RT‐qPCR analysis. In the 3D model, after 216 h of treatment, cirsimarin reduced cell viability (≥ 40 μM) and spheroid area (≥ 80 μM) while antimigratory effects were observed only in the highest concentration evaluated (160 μM). These findings could indicate a potential reduction in lung tumor growth and metastasis, warranting further investigation, particularly of the antimetastatic effect of this flavone.

In 2D culture, the cirsimarin (i) decreased cell viability in both assays, (ii) induced apoptosis, (iii) reduced clonogenicity, (iv) prevented cell migration, (v) induced DNA damage, and (vi) negatively modulated gene expression. In the 3D model, reductions in spheroid viability, area, and cell migration were also observed.