Characterization of a Novel Dermal Fibrosis Model Induced by Areca Nut Extract that Mimics Oral Submucous Fibrosis.

Oral submucous fibrosis (OSF) is an oral potentially malignant disorder and areca quid chewing is the main etiological factor. However, the molecular mechanism underlying OSF remains unclear, partly due to the lack of an appropriate animal model. The present study aimed to establish and characterize an animal model of areca nut extract (ANE)-induced skin fibrosis that mimics OSF. Mice were divided into 4 groups: the control group; the bleomycin group; and the ANE10 and ANE20 groups, which received 10mg/ml and 20mg/ml subcutaneous (SC) injection of ANE, respectively. Skin fibrosis was evaluated by histological analyses. Additionally, the expression levels of the fibrotic marker genes were determined by immunohistochemical staining and immunoblotting. ANE administration significantly increased dermal thickness and collagen deposition compared with the control group. Moreover, ANE induced the expression of the fibrotic marker genes alpha smooth muscle actin (α-SMA) and connective tissue growth factor (CTGF) in the skin lesions. The SC injection of ANE successfully induced skin fibrosis, exhibiting characteristics similar to those of OSF. This model may facilitate future studies of the mechanism underlying OSF.

Chemopreventive effect of Toona sinensis leaf extract on 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch squamous cell carcinogenesis.

OBJECTIVE: Toona sinensis leaf extract (TSL) has been shown to have anti-tumor effects on cancer cell lines. This study aimed to investigate the chemopreventive potential and the underlying mechanism of TSL during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. METHODS: One hundred hamsters were divided into control (n=30), carcinogenic (n=20), preventive (n=42), and therapeutic (n=8) groups. The animals in carcinogenic and preventive groups were administered reverse osmosis water (carcinogenic group) or TSL (1g/kg bw) (preventive group) by gavage daily for 4 weeks, and their bilateral pouches were painted with a 0.5% DMBA solution for 4, 9, and 12 weeks. The animals in the therapeutic group were treated with DMBA for 12 weeks prior to TSL administration for 4 weeks. Expression levels of survivin, X chromosome-linked inhibitor of apoptosis (XIAP), proliferating cell nuclear antigen (PCNA), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) proteins were analyzed by immunohistochemistry. Apoptotic activity was examined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method, cytochrome C, and poly (ADP-ribose) polymerase (PARP). RESULTS: In the preventive group, the results showed significant decreases not only in the incidences of squamous cell carcinoma (SCC) (50%) and epithelial dysplasia (62.5%) but also in the tumor number, tumor volume, tumor burden, and the severity of dysplastic lesions. The down-regulation of survivin, XIAP, PCNA, iNOS, and COX-2 proteins and the increased apoptotic activity indicated anti-proliferative and apoptosis-inducing abilities of TSL on DMBA-induced HBP carcinogenesis. CONCLUSIONS: The results suggested that TSL might be a promising candidate for the prevention of oral cancer.

Anti-invasion and anti-tumor growth effect of doxycycline treatment for human oral squamous-cell carcinoma--in vitro and in vivo studies.

Regional lymph node and distant organ metastasis of oral squamous-cell carcinoma (OSCC) has been associated with increased production of matrix metalloproteases (MMPs), and scientific data showed that doxycycline (Dox) could down-regulate the expression of MMPs. The objective of this study was to evaluate the effect of Dox on the expression of MMPs in vitro using the SCC-15 cell line and in vivo SCC-15 xenografted nude mice. SCC-15 cells maintained under distinct culture conditions expressed high levels of pro-MMP-2 and pro-MMP-9; however, as determined by zymography and Western blot analysis, Dox significantly reduced the production of pro-MMP-2 and pro-MMP-9 after 24h of treatment in a dose-dependent manner (2.5-40 microg/ml). Dox (10 microg/ml) decreased the expression of MMP-9 mRNA but did not alter the level of MMP-2 mRNA after 24h of treatment. In addition, this drug significantly inhibited the invasive and migration activities of SCC-15 cells in vitro (>75% inhibition at 10 microg/ml). On the other hand, daily administration of Dox (3mg/mice) restrained tumor growth in SCC-15 xenografted nude mice, with an inhibition rate of 85.6%. Compared with the control group (treated with normal saline), MMP-9 mRNA levels in the fresh tumor tissue decreased upon Dox treatment (P<0.01) while MMP-2 mRNA levels were unchanged. In conclusion, reduced expression of MMP-9 at the transcriptional level and MMP-2 at the post-transcriptional level caused by Dox was found to be associated with decreased invasion of oral SCC in vitro. Moreover, Dox exerted a significant suppressive effect on tumor growth in an in vivo nude mice model. Taken together, these results, to our knowledge, may first imply that Doxycycline has an adjuvant therapeutic effect on OSCC that is associated with inhibition of MMPs expression.