Licochalcone A suppresses migration and invasion of human hepatocellular carcinoma cells through downregulation of MKK4/JNK via NF-κB mediated urokinase plasminogen activator expression.

Jen-Pi Tsai,Yi-Hsien Hsieh,Da-Tian Bau,Shun-Fa Yang,Pei-Ching Hsiao,Chu-Liang Ling,Chun-Li Pai,Shu-Ching Hsieh,Xin-Yuan Guan

doi:10.1371/journal.pone.0086537

Abstract

Hepatocellular cell carcinoma (HCC) is one of the most commonly diagnosed cancers worldwide and in Taiwan. Chemoprevention of cancer with dietary bioactive compounds could potentially reverse, suppress, or prevent cancer progression. Licochalcone A (LicA) is a characteristic chalcone of licorice, which is the root of Glycyrrhiza inflate. It had been reported that LicA has anti-inflammatory, anti-microbial, and anti-tumor properties. However, the effects of LicA on the migration and invasion of human HCC cells have not yet been reported. In the present study, it was found that LicA inhibits the migratory and invasion ability of SK-Hep-1 and HA22T/VGH cells in a dose-dependent manner, as assessed by the cell migration and Matrigel cell invasion assay. Using casein zymography, Western blotting, reverse transcriptase polymerase chain reaction, and an immunofluorescence assay, it was found that LicA induces a dose-dependent inhibition of uPA activity and expression, as well as reduces mRNA levels in SK-Hep-1 and HA22T/VGH cells. LicA was also found to inhibit the expression of phosphor-JNK and phosphor-MKK4 in SK-Hep-1 cells. Furthermore, LicA significantly decreased uPA levels in SP600125-treated or si-MKK4-transfected cells alongside a marked reduction in cell migration and invasion, which supports the notion that an inhibition of MKK4/JNK results in anti-metastatic effects. Moreover, LicA inhibited the expression of nuclear NF-κB, as well as the binding ability of NF-κB to the uPA promoter. These findings further our understanding of the role of LicA in suppressing tumor metastasis and its underlying molecular mechanisms, as well as suggest that LicA may be a promising anti-metastatic agent.

Highlights

Hepatocellular cell carcinoma (HCC) has been diagnosed in more than half a million people worldwide
Prior to investigating the cytotoxic effects of Licochalcone A (LicA) on two human HCC cells (SK-Hep-1 and HA22T/VGH) and rat normal hepatic cells with an MTT assay, we found that LicA does not have a dose- and time-dependent inhibitory effect on the growth of the human HCC cells (Fig. 1A, 1B) and normal hepatic cells (Fig. 1C)
These findings suggest that treatment with LicA inhibits human HCC cell migration and invasion

Summary

Introduction

Hepatocellular cell carcinoma (HCC) has been diagnosed in more than half a million people worldwide. Risk factors for the development of HCC include viral hepatitis (i.e., hepatitis B virus and hepatitis C virus), alcoholic liver disease, potentially nonalcoholic fatty liver disease, and some other rare etiologies, such as hereditary hemochromatosis, autoimmune hepatitis, and Wilson’s disease [1]. Studies have reported that the development of HCC could be caused by multiple risk factors rather than a single risk factor, and that after HCC develops, distant metastasis becomes an importance index of prognosis [2,3]. Chemoprevention of cancer with dietary bioactive compounds may potentially reverse, suppress, or prevent cancer progression [4,5]. It is important to inhibit the spread of tumor cells to prevent the development of metastasis. Many dietary bioactive components have shown promising anti-cancer activities with little or no toxicity to normal cells [6]

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Results

Conclusion