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

Jung Chun Liao¹, Kun Tsung Lee², Bang Jau You³

¹School of Pharmacy, China Medical University, Taichung, Taiwan.

Food & Nutrition Research

|December 25, 2015

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.

Insights

Trigonelline, found in foods like snow peas, effectively inhibits liver cancer cell migration by downregulating key signaling pathways and antioxidant enzymes. This suggests its potential as a dietary chemopreventive agent against cancer progression.

Area of Science:

Biochemistry
Molecular Biology
Oncology

Background:

Trigonelline, present in dietary plants and coffee, exhibits anti-carcinogenic properties.
Epidemiological studies link coffee consumption to reduced risk of liver diseases like cirrhosis and hepatocellular carcinoma.
Investigating trigonelline as a chemopreventive agent for cancer is warranted.

Purpose of the Study:

To investigate the chemopreventive potential of trigonelline against liver cancer progression.
To analyze trigonelline content in dietary sources like snow pea (Pisum sativum).
To elucidate the molecular mechanisms underlying trigonelline's anti-cancer effects on hepatoma cells.

Main Methods:

High-performance liquid chromatography (HPLC) for trigonelline quantification in Pisum sativum.
Wound migration assays to assess hepatoma cell (Hep3B) migratory activity.
Western blotting, real-time RT-PCR, and flow cytometry to analyze protein expression, gene expression, and cell cycle, respectively.
Native gel electrophoresis to determine the activity of antioxidant enzymes.

Main Results:

Pisum sativum contains a significant amount of trigonelline.
Trigonelline (75-100 µM) effectively inhibited Hep3B cell migration without impacting proliferation.
Trigonelline treatment downregulated Raf/ERK/Nrf2 signaling, MMP-7 gene expression, and the activity of antioxidant enzymes (SOD, catalase, glutathione peroxidase).

Conclusions:

Trigonelline inhibits Hep3B cell migration via the PKCα and Raf/ERK/Nrf2 signaling pathways and MMP-7 gene expression.
The chemopreventive effect involves the downregulation of Nrf2-dependent antioxidant enzymes.
Trigonelline, abundant in daily foods like snow peas, shows promise as a dietary chemopreventive agent for cancer prevention.

Read full analysis

Keywords:

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

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