Effect Of Benzyl-isothiocyanate On The Growth Of Squamous Cells Carcinoma Cells In Human Tongue

Background:Benzyl-isothiocyanate (BITC) is one of the naturally isothiocyanate compounds (ITCs), which is richest in the degradation products of glucosinolates in horseradish, mustard, cabbage, cauliflower, water celery and other cruciferous vegetables. According to the studies, BITC has a high degree of biological activities, such as cancer prevention ability, anti-inflammatory, inhibiting platelet aggregation. Epidemiological studies show that regular consumption of cruciferous vegetables can obviously reduce the risk of lung cancer, stomach cancer, prostate cancer, bowel cancer. It is well known that ITCs have shown unique cancer preventive properties. Evidence is accumulating to indicate that ITC can suppress proliferation of cancer cells in culture by causing apoptosis and/or cell cycle arrest. However, not much is known about BITC on inhibiting oral cancer cell growth on and the molecules involved in regulation. It attracted a great deal of attention due to its remarkable chemoprevention activity and can be used as a nutrition supplement. Here, we provide experimental evidence to the effect of BITC on the growth of squamous cell carcinoma cell lines in human tongue (hTSCC). Furthermore, the present study will discuss whether BITC may induce tongue squamous cell carcinoma cell lines apoptosis through Caspase 3 dependent mechanism.Methods:hTSCC including CAL-27 and SCC-9 were cultured to investigate the effect of proliferation and apoptosis induced by BITC. The inhibitory effects of BITC on human tongue squamous cell carcinoma cell lines CAL-27 in vitro were detected by MTS assay, cell morphological changes, respetely cell growth curve, cell cycle distribution was analyzed by flow cytometry (FCM). Apoptosis was determined by Annexin V-FITC staining. Caspase 8, Caspase 9 and Caspase 3 protein expression were measured by Western Blot analysis.Results:The results showed that BITC inhibited hTSCC proliferation in concertration dependent manner. When cells were treated with 7.5μM BITC for 48 h, the proliferation inhibition rate of hTSCC arrived at 56.2%, compared with the DMSO control group (P<0.05). Also in Group 7.5μM BITC for 48 h, the cell morphology observed by inverted microscope exhibited many changes including blebbing, loss of cell membrane asymmetry and attachment, cell shrinkage, nuclear fragmentation. Meanwhile, BITC induced cell apoptosis in a dose dependent manner. When hTSCCs were treated with 2.5μM,5μM,7.5μM BITC for 48 h, the rate of apoptosis were 3.7 %,6.2%,8.6%, separately. BITC arrested hSTCC cell cycle at G2/M phase. Furthermore, the protein level of Caspase 8, Caspase 9, Caspase 3, were increased significantly in BITC-treated SCC-9 cells.Conclusions:This study demonstrates that dietary BITC has anti-proliferative activity against human tongue squamous cell carcinoma cells and serves as apoptosis inductors. The data suggest that Caspase 8,9 and 3 pathways have a critical role in BITC-induced apoptosis. The study suggests BITC could be chemotherapeutic agent in hSTCC.