Suppression of inflammation-induced lung cancer cells proliferation and metastasis by exiguaflavanone A and exiguaflavanone B from Sophora exigua root extract through NLRP3 inflammasome pathway inhibition

Punnida Arjsri; Kamonwan Srisawad; Warathit Semmarath; Sonthaya Umsumarng; Lapamas Rueankham; Aroonchai Saiai; Methee Rungrojsakul; Trinnakorn Katekunlaphan; Songyot Anuchapreeda; Pornngarm Dejkriengkraikul

doi:10.3389/fphar.2023.1243727

Suppression of inflammation-induced lung cancer cells proliferation and metastasis by exiguaflavanone A and exiguaflavanone B from Sophora exigua root extract through NLRP3 inflammasome pathway inhibition

Front Pharmacol. 2023 Nov 10:14:1243727. doi: 10.3389/fphar.2023.1243727. eCollection 2023.

Authors

Punnida Arjsri^{1

2}, Kamonwan Srisawad^{1

2}, Warathit Semmarath^{1

3}, Sonthaya Umsumarng^{4

5}, Lapamas Rueankham⁶, Aroonchai Saiai⁷, Methee Rungrojsakul⁸, Trinnakorn Katekunlaphan⁹, Songyot Anuchapreeda⁶, Pornngarm Dejkriengkraikul^{1

2

4}

Affiliations

¹ Department of Biochemistry, Faculty Medicine, Chiang Mai University, Chiang Mai, Thailand.
² Anticarcinogenesis and Apoptosis Research Cluster, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
³ Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, Thailand.
⁴ Center for Research and Development of Natural Products for Health, Chiang Mai University, Chiang Mai, Thailand.
⁵ Division of Veterinary Preclinical Sciences, Department of Veterinary Biosciences and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.
⁶ Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
⁷ Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai, Thailand.
⁸ Department of Traditional Chinese Medicine, Faculty of Science, Chandrakasem Rajabhat University, Bangkok, Thailand.
⁹ Department of Chemistry, Faculty of Science, Chandrakasem Rajabhat University, Bangkok, Thailand.

Abstract

Objective: Non-small cell lung cancer (NSCLC) is recognized for its aggressive nature and propensity for high rates of metastasis. The NLRP3 inflammasome pathway plays a vital role in the progression of NSCLC. This study aimed to investigate the effects of S. exigua extract and its active compounds on NLRP3 regulation in NSCLC using an in vitro model. Methods: S. exigua was extracted using hexane, ethyl acetate and ethanol to obtain S. exigua hexane fraction (SE-Hex), S. exigua ethyl acetate fraction (SE-EA), and S. exigua ethanol fraction (SE-EtOH) respectively. The active compounds were identified using column chromatography and NMR analysis. A549 cells were primed with lipopolysaccharide (LPS) and adenosine triphosphate (ATP) for activated NLRP3 inflammasome. The anti-inflammatory properties were determined using ELISA assay. The anti-proliferation and anti-metastasis properties against LPS-ATP-induced A549 cells were determined by colony formation, cell cycle, wound healing, and trans-well migration and invasion assays. The inflammatory gene expressions and molecular mechanism were determined using RT-qPCR and Western blot analysis, respectively. Results: SE-EA exhibited the greatest anti-inflammation properties compared with other two fractions as evidenced by the significant inhibition of IL-1β, IL-18, and IL-6, cytokine productions from LPS-ATP-induced A549 cells in a dose-dependent manner (p < 0.05). The analysis of active compounds revealed exiguaflavanone A (EGF-A) and exiguaflavanone B (EGF-B) as the major compounds present in SE-EA. Then, SE-EA and its major compound were investigated for the anti-proliferation and anti-metastasis properties. It was found that SE-EA, EGF-A, and EGF-B could inhibit the proliferation of LPS-ATP-induced A549 cells through cell cycle arrest induction at the G0/G1 phase and reducing the expression of cell cycle regulator proteins. Furthermore, SE-EA and its major compounds dose-dependently suppressed migration and invasion of LPS-ATP-induced A549 cells. At the molecular level, SE-EA, EGF-A, and EGF-B significantly downregulated the mRNA expression of IL-1β, IL-18, IL-6, and NLRP3 in LPS-ATP-induced A549 cells. Regarding the mechanistic study, SE-EA, EGF-A, and EGF-B inhibited NLRP3 inflammasome activation through suppressing NLRP3, ASC, pro-caspase-1(p50 form), and cleaved-caspase-1(p20 form) expressions. Conclusion: Targeting NLRP3 inflammasome pathway holds promise as a therapeutic approach to counteract pro-tumorigenic inflammation and develop novel treatments for NSCLC.

Keywords: NLRP3 inflammasome pathway; Sophora exigua; cancer progression and metastasis; exiguaflavanone; non-small cell lung cancer.

Grants and funding

This research project was supported by Fundamental Fund 2023, Chiang Mai University (Grant number FF66/046). This research is partially supported by Chiang Mai University, the Anticarcinogenesis and Apoptosis Research Cluster and the Center for Research and Development of Natural Products for Health, Chiang Mai University.