Raf/ERK/Nrf2 signaling pathway and MMP-7 expression involvement in the trigonelline-mediated inhibition of hepatocarcinoma cell migration

Jung Chun Liao; Kun Tsung Lee; Bang Jau You; Chia Lin Lee; Wen Te Chang; Yang Chang Wu; Hong-Zin Lee

doi:10.3402/fnr.v59.29884

Jung Chun Liao School of Pharmacy, China Medical University, Taichung, Taiwan
Kun Tsung Lee Department of Oral Hygiene, College of Dental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
Bang Jau You School of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung, Taiwan
Chia Lin Lee Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan
Wen Te Chang School of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung, Taiwan
Yang Chang Wu School of Pharmacy, China Medical University, Taichung, Taiwan
Hong-Zin Lee School of Pharmacy, China Medical University, Taichung, Taiwan

DOI: https://doi.org/10.3402/fnr.v59.29884

Keywords: Trigonelline, Hepatocarcinoma Hep3B cells, Migration, Pisum sativum, Chemotherapeutic agent, Raf/ERK/Nrf2 signaling pathway, MMP-7

Abstract

Background: Trigonelline occurs in many dietary food plants and has been found to have anti-carcinogenic activity. Trigonelline is also found in coffee which is one of the most widely consumed beverages. Many epidemiological studies have reported that coffee consumption has an inverse relationship with the risk of cirrhosis or hepatocellular carcinoma. It would be interesting to investigate whether trigonelline is an ideal chemoprevent agent to prevent cancer progression.

Methods: The protein expression was performed by western blotting. The trigonelline content in snow pea (Pisum sativum) was analyzed by high-performance liquid chromatography (HPLC). The migratory activity of human hepatocarcinoma cells (Hep3B) was assessed by using a wound migration assay. The percentage of each phase in the cell cycle was analyzed on a FACScan flow cytometer. Gene expression was detected by real-time reverse transcriptase-polymerase chain reaction techniques. Native gel analysis was performed to analyze the activity of superoxide dismutase (SOD), catalase and glutathione peroxidase.

Results: According to the data of HPLC analysis, P. sativum, which is a popular vegetable, has relatively high content of trigonelline. Our findings suggest that trigonelline is an efficient compound for inhibiting Hep3B cell migration. Trigonelline inhibited the migration of hepatoma cells at concentrations of 75–100 µM without affecting proliferation. Raf/ERK/Nrf2 protein levels and further downstream antioxidative enzymes activity, such as SOD, catalase, and glutathione peroxidase, significantly decreased after treatment with 100 µM of trigonelline for 24 h. The migration inhibition of trigonelline is also related to its ability to regulate the matrix metalloproteinases 7 (MMP-7) gene expression.

Conclusions: In this study, protein kinase Cα (PKCα) and Raf/ERK/Nrf2 signaling pathway and MMP-7 gene expression were involved in the trigonelline-mediated migration inhibition of Hep3B cells. We also demonstrated that trigonelline inhibits Hep3B cell migration through downregulation of nuclear factor E2-related factor 2–dependent antioxidant enzymes activity. This study analyzed the trigonelline content in a popular vegetable, snow pea, as a representative proof to prove that trigonelline is often found in the daily intake of food. Our finding suggested that trigonelline should be a useful chemopreventive agent derived from the daily intake of food to prevent cancer progression.

Keywords: trigonelline; hepatocarcinoma Hep3B cells; migration; Pisum sativum; chemotherapeutic agent; Raf/ERK/Nrf2 signaling pathway; MMP-7

(Published: 22 December 2015)

Citation: Food & Nutrition Research 2015, 59: 29884 - http://dx.doi.org/10.3402/fnr.v59.29884

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