Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC)

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Guibitang, a traditional herbal medicine, induces apoptotic death in A431 cells by regulating the activities of mitogen-activated protein kinases


Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are commonly referred to as non-melanoma skin cancers [1,2]. BCC is a slow-growing cancer that does not usually metastasize. Similarly, SCC is frequently localized without evidence of blood-born metastasis, making direct treatment of the tumor straightforward. However, SCC is the sixth most common cancer worldwide, and its incidence has increased dramatically at multiple sites in the body, including the head and neck, cervix, and lung [3,4]. Accordingly, it is necessary to develop novel effective chemopreventive agents to inhibit the development of SCC.

Guibitang (GBT), known as ‘Kihi-to’ in Japan and ‘Gui-Pi-Tang’ in China, is a traditional medicine and herbal formula that has been used for several hundred years, predominantly to treat insomnia, amnesia, palpitations, anxiety, fatigue, poor appetite, and depression [5]. Recent studies have reported the specific bioactivities of GBT, which include immune regulation [6], antioxidant effects [7], and protective effect of the gastric mucosa [8].

GBT is an aqueous polyherbal formulation that contains 12 herbs: Angelica gigas Nakai, Dimocarpus longan, Zizyphus jujuba Miller (seed), Polygala tenuifolia, Panax ginseng, Astragalus membranaceus, Atractylodes ovate, Poria cocos, Inula helenium, Glycyrrhiza glabra, Zingiber officinale, and Zizyphus jujuba Miller (Fructus). GBT also regulates chronic fatigue syndrome-associated cytokine production, whereas the addition of Gardenia jasminoides, Paeonia suffruticosa, and Bupleurum falcatum to GBT improves palliative care in patients undergoing chemotherapy for ovarian cancer [9]. Although it has been shown that adding several herbs to GBT results in anti-cancer effects against gynecological or lung cancer, the molecular mechanisms behind these effect of GBT remain unclear.

Tumorigenesis is caused by unregulated growth of cells resulting from DNA damage, mutations of functional genes, dysregulation of the cell cycle, and loss of apoptotic function [10]. Therefore, regulating the induction of apoptosis by modulating cell growth and survival-related signaling pathways is a common and major target for cancer therapies [11]. Among several signaling pathways in cancer cells, mitogen-activated protein kinase (MAPK) signals including extracellular signal-regulated kinases (ERK), p38 kinases, and c-Jun N-terminal kinases (JNK), take an important role in cell death and survival [12]. The regulation of ERK activation is induced by conditions of stress such as some agents and oxidant injury, which plays a major role in regulating cell growth and differentiation [13]. JNK and p38 are activated in response to several stress signals including tumor necrosis factor and hyperosmotic condition, which is associated with induction of apoptosis [14].

GBT showed cytotoxic activity against three different squamous cell carcinoma, especially on A431 cells. GBT induced the apoptosis through activating the caspase-8 in A431 cells. Inhibition of A431 cell growth by GBT was caused by G1-phase arrest through regulating proteins associated with cell cycle progression, such as cyclin D1, p21, and p27. Furthermore, GBT regulated the activation of mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase (ERK), p38 and c-Jun NH2-terminal kinase (JNK), and activated p53, a tumor suppressor protein. In MAPKs inhibitor study, inhibitors respectively blocked GBT-induced cell viability, indicating that MAPKs signals play critical role in cell death caused by GBT. In vivo xenografts, daily oral administration of 600 mg/kg GBT efficiently suppressed the tumorigenic growth of A431 cells without side effects such as loss of body weight and change of toxicological parameters compared to vehicle.

In the present study, Yim et al., (2014) evaluated whether GBT shows the anti-cancer effect in A431 human squamous carcinoma cells, which demonstrated that GBT induces apoptosis of cancer cells specifically, as an inhibition of the cell growth via regulating MAPK signaling pathway in A431 cells.

GBT decreases cell viability in A431 human squamous carcinoma cells
Six different human cancer cell lines (A431 [squamous], AGS [stomach], HeLa [cervical], Caki-1 [kidney], SK-Hep-1 [liver], and HCT116 [colon]) were treated with 500 ?g/mL GBT for 48 h, and cell viability was assessed by an MTT assay. Although most cell lines were unaffected, the viability of A431 cells was inhibited >35% by treatment with GBT (Figure 2A). Therefore, subsequent tests focused on A431 cells.

To further define the inhibitory action of GBT on SCCs, the suppression of cell growth by GBT on three different SCC lines (SCC12, SCC13, and A431) was evaluated. As shown in Figure 2B, treatment with 500 and 1000 ?g/mL GBT for 48 h reduced the viability of A431 cells by 35% and 52%, respectively. Treatment of SCC13 cells with 1000 ?g/mL GBT also inhibited the cell growth by ~30% although these effects were not as potent as those observed in A431 cells.

In contrast, the viability of SCC12 cells was not affected significantly by GBT. The potential cytotoxic effect of GBT on normal cells was assessed using normal human HaCaT keratinocytes and mouse primary liver cells. HaCaT cells were unaffected by GBT under the same conditions that were cytotoxic to A431 cells (Figure 2C). In addition, no cytotoxic effects on primary liver cells were observed by treatment with 500 ?g/mL or 1000 ?g/mL GBT. Instead, GBT weakly increased the viability of liver primary cells in a dose- and time-dependent manner. These results suggest that GBT has cancer-specific cytotoxic effect on A431 cells, without affecting normal cells.

Yim N-H, Kim A, Liang C et al. BMC Complementary and Alternative Medicine 2014, 14:344 doi:10.1186/1472-6882-14-344

1. Diepgen TL, Mahler V: The epidemiology of skin cancer. Br J Dermatol 2002, 146(Suppl 61):1-6.
2. Weinstein MC, Brodell RT, Bordeaux J, Honda K: The art and science of surgical margins for the dermatopathologist. Am J Dermatopathol 2012, 34(7):737-745.
3. Sauter ER, Herlyn M, Liu SC, Litwin S, Ridge JA: Prolonged response to antisense cyclin D1 in a human squamous cancer xenograft model. Clin Cancer Res 2000, 6(2):654-660.
4. Trakatelli M, Ulrich C, del Marmol V, Euvrard S, Stockfleth E, Abeni D: Epidemiology of nonmelanoma skin cancer (NMSC) in Europe: accurate and comparable data are needed for effective public health monitoring and interventions. Br J Dermatol 2007, 156(Suppl 3):1-7.
5. Tohda C, Ichimura M, Bai Y, Tanaka K, Zhu S, Komatsu K: Inhibitory effects of Eleutherococcus senticosus extracts on amyloid beta(25–35)-induced neuritic atrophy and synaptic loss. J Pharmacol Sci 2008, 107(3):329-339.
6. Busta I, Xei HS, Kim MS: The use of Gui-Pi-Tang in small animals with immune-mediated blood disorders. J Vet Clin 2009, 26:181-184.
7. Kang IH, Lee I, Han SH, Moon BS: Effects of Gwibitang on glutamate-induced apoptosis in C6 glial cells. J Korean Orient Med 2001, 22:45-57.
8. Kim HJ, Choi JH, Lim SW: The defensive effect of Keuibi-tang on the gastric mucous membrane of mouse injured by stress and ethanol. J Orient Med 2003, 24:155-168.
9. Ikeda A, Higashio S, Ushiroyama T: Experience with administration of kamikihito with chemotherapy and palliative care in patients with gynecologic cancer. Recent Prog Kampo Med Obstet Gynecol 2003, 20:152-155.
10. Kundoor V, Zhang X, Bommareddy A, Khalifa S, Fahmy H, Dwivedi C: Chemopreventive effects of sarcotriol on ultraviolet B-induced skin tumor development in SKH-1 hairless mice. Marine Drugs 2007, 5(4):197-207.
11. Sarfaraz S, Adhami VM, Syed DN, Afaq F, Mukhtar H: Cannabinoids for cancer treatment: progress and promise. Cancer Res 2008, 68(2):339-342.
12. Hamamura K, Goldring MB, Yokota H: Involvement of p38 MAPK in regulation of MMP13 mRNA in chondrocytes in response to surviving stress to endoplasmic reticulum. Arch Oral Biol 2009, 54(3):279-286.
13. Fan M, Chambers TC: Role of mitogen-activated protein kinases in the response of tumor cells to chemotherapy. Drug Resist Updat 2001, 5:253-267.
14. Dent P, Grant S: Pharmacologic interruption of the mitogen-activated extracellular-regulated kinase/mitogen-activated protein kinase signal transduction pathway: potential role in promoting cytotoxic drug action. Clin Cancer Res 2001, 7:775-783.

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