Wogonin, an active ingredient of Chinese herb medicine Scutellaria baicalensis, inhibits the mobility and invasion of human gallbladder carcinoma GBC-SD cells by inducing the expression of maspin
Graphical abstract
Wogonin, an active ingredient of Chinese herb medicine Scutellaria baicalensis, inhibits the mobility and invasion of human gallbladder carcinoma GBC-SD cells by inducing the expression of maspin.
Introduction
Tumor growth, invasion, and metastasis are multistep and complex processes that include cell division and proliferation, proteolytic digestion of the extracellular matrix, cell migration through basement membranes to reach the circulatory system, and remigration and growth of tumors at the metastatic sites (Klein, 2008). It is firmly established that matrix metalloproteinase (MMP)-2 and MMP-9 secretion from cancerous cells leads to hydrolysis of extracellular matrix, thus enabling cells to break out of their primary site into the circulation and form the secondary metastatic sites. Invasiveness inhibitors may act by down-modulating extracellular-barrier degrading proteinases such as MMP-2 and MMP-9 (Nozaki et al., 2003). Activation of extracellular regulated protein kinase 1/2 (ERK1/2) has been shown to be the mechanism for promoting the production of MMPs (Beshir et al., 2010), which are important for cell proliferation, invasion, and neovascularization. It has been shown that inhibition of ERK1/2 activation may lead to a reduction in the expression of MMP-2, as well as in the invasion of tumor cells (Lev et al., 2004).
Maspin, a member of the serine protease inhibitor family, has been shown to be a potent metastasis suppressor (Bailey et al., 2006, Khalkhali-Ellis, 2006, Lockett et al., 2006). The ectopic expression of maspin suppresses the invasive potential of breast cancer cells in vitro and the growth and metastasis of prostate and breast cancers in a mouse model (Zou et al., 1994, Shi et al., 2001). Maspin is expressed ubiquitously in multiple tissues and normal epithelial cells but reduced in tumor cells (Zou et al., 1994, Khalkhali-Ellis, 2006). Thus, inducing maspin expression in premalignant and malignant cells may be an effective strategy for cancer prevention and treatment.
Scutellaria (HUANG QIN, Lamiaceae), which includes about 350 species commonly known as skullcaps, is widespread in Europe, the United States and East Asia (Shang et al., 2010). A traditional Chinese medicine Scutellaria baicalensis is prescribed for the treatment of a variety of inflammatory diseases and tumors in clinic in China (Shang et al., 2010). The crude extract of Scutellaria baicalensis has been reported to exert anti-cancer effect against some human tumor cells (Ye et al., 2002, Kumagai et al., 2007). Wogonin, a main active ingredient of Chinese herb medicine Scutellaria baicalensis, exhibits a variety of pharmacological activities including antioxidant (Gao et al., 1999), anti-inflammatory (Chi et al., 2001), anxiolytic (Hui et al., 2002), anti-hepatitis (Guo et al., 2007), anticonvulsant (Park et al., 2007), anti-angiogenesis (Lu et al., 2008), neuroprotective (Lee et al., 2003, Lim et al., 2010), anticancer (Wang et al., 2006, Baumann et al., 2008, Li-Weber, 2009) activities. Recent study has demonstrated that wogonin at the dose of more than 20 μM could effectively inhibit the proliferation and induce the apoptosis in several cancer cell lines in vitro (Lee et al., 2008, Zhang et al., 2008). However, little is known about the action of wogonin on tumor cells at the sub-dose of apoptosis-inducing effect. Here we found that wogonin at the dose of 1–10 μM, which did not induce apoptosis, significantly inhibited the mobility and invasion activity of human gallbladder carcinoma GBC-SD cells. Furthermore, the metastasis suppressor maspin had been confirmed as the downstream target of wogonin. Our data provide novel insights into the chemoprotective effect of wogonin, a main active ingredient of Chinese medicine Scutellaria baicalensis, against highly metastatic tumor.
Section snippets
Cell culture
All tumor cells used in the present study were maintained in HG-Dulbecco's modified Eagle's medium (4.5 g/l glucose; Invitrogen) supplemented with 10% fetal calf serum (FCS, Invitrogen) plus 2 mM glutamine and 50 U/ml penicillin. Human gallbladder carcinoma GBC-SD, human prostate carcinoma LNCaP cells and human U-937 leukemia cells were purchased from Cell Bank of Shanghai Institute of Biochemistry and Cell Biology (Chinese Academy of Sciences, China) and they were grown at 37 °C in a 5% (v/v) CO2
Wogonin inhibited the mobility and invasive potency of human gallbladder carcinoma GBC-SD cells at the sub-dose of apoptosis-inducing effect
As shown in Fig. 1A, wogonin at the dose of more than 10 μM killed human gallbladder carcinoma GBC-SD cells, human prostate carcinoma LNCaP cells and human U-937 leukemia cells in a concentration-depedent manner. However, wogonin at the concentrations of 1–10 μM did not affect the survival of cells. In addition, wogonin at the dose of 1–10 μM did not induce apoptosis of GBC-SD cells (Fig. 1B).
To examine the effect of wogonin at the sub-dose of apoptosis-inducing effect (1–10 μM) on cancer cell
Discussion
The highlight of this work is the revelation that in the 1–10 μM range wogonin does not induce apoptosis but significantly inhibits the mobility and invasion activity of human gallbladder carcinoma GBC-SD cells by upregulating the metastasis suppressor maspin. Previously, wogonin has been reported to effectively inhibit the proliferation and induce the apoptosis at the dosages of more than 10 μM in several cancer cell lines (Lee et al., 2008, Zhang et al., 2008). We also found that wogonin at the
Conclusion
In conclusion, wogonin upregulates the expression of metastasis suppressor maspin, which is involved in anti-invasive activity of wogonin in human gallbladder carcinoma GBC-SD cells. The present study provides a novel insight into the anticancer activity of wogonin, a main active ingredient of Chinese medicine Scutellaria baicalensis.
Conflict of interest statement
The authors have no conflicts of interest.
Acknowledgements
This study was supported by National Natural Science Foundation of China (Nos. 30640060, 30872502, 30972918), Natural Science Foundation of Zhejiang Province for Outstanding Young Researcher Groups (No. R2080452) and Foundation of Medical School, Shanghai Jiao Tong University (No. 09XJ21012).
Glossary
- DMEM
- Dulbecco's modified Eagle medium
- ERK1/2
- extracellular regulated protein kinase 1/2
- FCS
- fetal calf serum
- FITC
- fluorescein isothiocyanate
- MMP
- matrix metalloproteinase
- MTT
- 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide
- PI
- propidium iodide
- RT-PCR
- reverse transcriptase-polymerase chain reaction
References (32)
- et al.
Wogonin preferentially kills malignant lymphocytes and suppresses T-cell tumor growth by inducing PLCgamma1- and Ca2+-dependent apoptosis
Blood
(2008) - et al.
Raf kinase inhibitor protein suppresses nuclear factor-kappaB-dependent cancer cell invasion through negative regulation of matrix metalloproteinase expression
Cancer Letter
(2010) - et al.
Free radical scavenging and antioxidant activities of flavonoids extracted from the radix of Scutellaria baicalensis Georgi
Biochimica et Biophysica Acta
(1999) - et al.
Anti-hepatitis B virus activity of wogonin in vitro and in vivo
Antiviral Research
(2007) - et al.
Anxiolytic effect of wogonin, a benzodiazepine receptor ligand isolated from Scutellaria baicalensis Georgi
Biochemical Pharmacology
(2002) - et al.
Scutellaria baicalensis, a herbal medicine: anti-proliferative and apoptotic activity against acute lymphocytic leukemia, lymphoma and myeloma cell lines
Leukemia Research
(2007) - et al.
Wogonin suppresses TNF-alpha-induced MMP-9 expression by blocking the NF-kappaB activation via MAPK signaling pathways in human aortic smooth muscle cells
Biochemical and Biophysical Research Communications
(2006) - et al.
Role of p53, PUMA, and Bax in wogonin-induced apoptosis in human cancer cells
Biochemical Pharmacology
(2008) - et al.
Wogonin induces differentiation and neurite outgrowth of neural precursor cells
Biochemical and Biophysical Research Communications
(2010) New therapeutic aspects of flavones: the anticancer properties of Scutellaria and its main active constituents Wogonin, Baicalein and Baicalin
Cancer Treatment Reviews
(2009)
Wogonin suppresses tumor growth in vivo and VEGF-induced angiogenesis through inhibiting tyrosine phosphorylation of VEGFR2
Life Sciences
Anticonvulsant effect of wogonin isolated from Scutellaria baicalensis
The European Journal of Pharmacology
Reactive oxygen species-mediated endoplasmic reticulum stress and mitochondrial dysfunction contribute to cirsimaritin-induced apoptosis in human gallbladder carcinoma GBC-SD cells
Cancer Letter
The genus Scutellaria an ethnopharmacological and phytochemical review
The Journal of Ethnopharmacology
Identification of metastasis-associated proteins involved in gallbladder carcinoma metastasis by proteomic analysis and functional exploration of chloride intracellular channel 1
Cancer Letter
Interaction of testisin with maspin and its impact on invasion and cell death resistance of cervical cancer cells
FEBS Letters
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2016, Tetrahedron LettersCitation Excerpt :The root extract was employed to treat hypertension, atherosclerosis, infections and fever, among other ailments.1,2 One of its major constituents is 5,7-dihydroxy-8-methoxyflavone, a.k.a. wogonin (1), which is known to have anti-inflammatory, antioxidant, antiangiogenic3 and antitumour properties.4 Despite its simple structure, chemical syntheses are few and far between.5–7
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These authors contributed equally to this work.