Cancer Letters

Cancer Letters

Volume 279, Issue 2, 8 July 2009, Pages 155-162
Cancer Letters

Berberine suppresses in vitro migration and invasion of human SCC-4 tongue squamous cancer cells through the inhibitions of FAK, IKK, NF-κB, u-PA and MMP-2 and -9

https://doi.org/10.1016/j.canlet.2009.01.033Get rights and content

Abstract

There is increasing evidence that urokinase-type plasminogen activator (u-PA) and matrix metalloproteinases (MMPs) play an important role in cancer metastasis and angiogenesis. Inhibition of u-PA and MMPs could suppress migration and invasion of cancer cells. Berberine, one of the main constituents of the plant Rhizoma coptidis, is a type of isoquinoline alkaloid, reported to have anti-cancer effects in different human cancer cell lines. There is however, no available information on effects of berberine on migration and invasion of human tongue cancer cells. Here, we report that berberine inhibited migration and invasion of human SCC-4 tongue squamous carcinoma cells. This action was mediated by the p-JNK, p-ERK, p-p38, IκK and NF-κB signaling pathways resulting in inhibition of MMP-2 and -9 in human SCC-4 tongue squamous carcinoma cells. Our Western blowing analysis also showed that berberine inhibited the levels of urokinase-plasminogen activator (u-PA). These results suggest that berberine down-regulates u-PA, MMP-2 and -9 expressions in SCC-4 cells through the FAK, IKK and NF-κB mediated pathways and a novel function of berberine is to inhibit the invasive capacity of malignant cells.

Introduction

It was reported that over 300,000 cases of oral and oropharyngeal cancer occur per year and occurrence is higher in males than in females [1]. Oral cancer cases and deaths are associated with individual predisposition such as specific genetic characteristics but lifestyle factors such as tobacco/bidi smoking, [2], [3] betel quid, [4] heavy alcohol drinking, [5] micronutrient deficiency [6] and human papillomavirus infection [7] play a role. Two or more of these factors can increase the risk of oral cancer [8], [9]. The betel quid chewing for example has a strong association with the occurrence of oral cancer in Taiwan. Based on reports from the People Health Bureau of Taiwan, approximately 6 per 100,000 die annually of oral cancer in Taiwan and oral cancer has been elevated to be the fourth most frequent cause of cancer death among males in Taiwan. Treatment options for oral cancer such as surgery, radiotherapy and chemotherapy have not been satisfactory. Therefore, it is important to identify new agents and novel targets for the treatment of oral cancer.

Berberine, a natural alkaloid found in natural plants, is anti-bacterial, [10] anti-oxidative, [11] anti-inflammatory, [12] anti-carcinogenic activity [13] and exerts anti-metastatic properties in non-small lung cancer cells [14]. It was reported that berberine suppressed cyclooxygenase-2 transcriptional and activator protein-1 activity in human colon cancer cells [15], [16] and DNA topoisomerase II [17]. Berberine induced cytotoxic activity in human leukemia U937 and murine melanoma B16 cells, [18] human prostate cancer cells [10] and human epidermoid carcinoma A431 cells [19].

In our laboratory, we have reported that berberine inhibited N-acetyltransferase activity in human colon tumor cells [20] and induced apoptosis in human oropharyngeal cancer HSC-3 cells [21] and inhibited rat vascular smooth muscle cell proliferation and migration in vitro[22]. Inhibition of berberine on the induction of migration and invasion in human tongue cancer cells has not been reported. The purpose of this study was to investigate effects of berberine on the induction of migration and invasion in human SCC-4 tongue squamous carcinoma cells. We show that berberine inhibited the migration and invasion of SCC-4 cells through the inhibition of MMP-2 and -9.

Section snippets

Materials and chemicals

Berberine, dimethyl sulfoxide (DMSO), trypan blue and triton X-100, pyruvate, penicillin G and streptomycin were obtained from Sigma Chemical. (St. Louis, MO, USA). Anti-MMP-2, anti-MMP-9, anti-FAK, anti-u-PA, anti-p-p38, anti-p-JNK, anti-p-ERK, anti-IKK, anti-NF-κB and anti-IκB were purchased from Santa Cruz Biotechnology. Materials and chemicals for electrophoresis were obtained from BioRad.

Cell culture

The human SCC-4 tongue squamous carcinoma cell line was purchased from the Food Industry Research and

Berberine decreased the percentages of viable SCC-4 cells

Effects of berberine on the percentage of viable SCC-4 cells were examined and quantified by trypan blue exclusion and flow cytometric analysis. It can be seen in Fig. 1 that berberine induced cytotoxicity and decreased the percentage of viable cells from 40% to 52% (P < 0.001) of 62.5 and 125 μM berberine at 48 h treatment but at 24 h treatment of berberin, only 125 μM induced a significant decrease of viable cells.

Berberine inhibited the migration of SCC-4 cells in vitro

Effects of berberine on cell migration were investigated using a wound-healing assay

Discussion

The anti-cancer effect of berberine has been well documented in many different types of human cancers [11], [12], [14], [15], [16], [17], [18], [19], [26]. In this study, our results demonstrated that berberine decreased the percentage of human SCC-4 tongue cancer viable cells in a dose-dependent manner, which is in agreement with our previous studies. However, actions of berberine on migration and invasion of SCC-4 cells and the associated signaling pathways have not been reported. In this

Conflict of Interest Statement

None declared.

Acknowledgments

This work was supported by Grant CMU97-094 from China Medical University, Taiwan and NIH grants AG-23524 and AG-18357, USA.

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