| Tongue squamous cell carcinoma is a high incidence disease in oral cancers. Surgery and radiotherapy are the main methods for the treatment of tongue cancer. However, the effect of radiotherapy is limited owing to the injury of healthy tissue and the resistance of tumor tissue. In clinical therapy, the use of radiosensitizer can improve the radiation effect on tumor cells. However, ideal radiosensitizer is nonexistence. Thus, Choose a good radiation sensitizer treatment has become a new way to optimize therapy of tongue cancer. Sodium Glycididazole is a radiosensitizer developed by Second Military Medical University. It is the first new structure of the 5-nitroimidazole compounds in the world. Sodium Glycididazole has solved the problom of radiosensitizer toxic side effects. At present, clinical studies confirmed the drug play a significant role in radiosensitization on nasopharyngeal carcinoma, lung cancer, cervical cancer, esophageal cancer and so on. Evodiamine is extracted from nearly ripe fruit of rutaceae evodia. In recent years, numerous studies have shown that EVO has anticancer activity on malignant melanoma, prostate cancer, liver cancer, thyroid cancer, stomach cancer and colon cancer and even multi-drug resistant tumor cells. At the present time,we have not see any reports about CMNa and EVO increase sensitizing effect of radiation on tongue cancer and any reports about EVO can increase sensitizing of radiation.This study is aimed to investigate radiosensitivity, migration and adhesion of CMNa and evodiamine with different doses radiotherapy on human tongue cancer Tca-8113 cell line. This study is divided into two parts.1. CMNa affects radiosensitivity and homogeneous adhesion of Tca-8113 cell line. Cells in logarithmic growth phase were used for experiment. Cells growth inhibition was assessed by MTT after treated with various concentrations and time. The results show surviaval rates of Tca-8113 cells were decreased as increase of CMNa concentration and time. The mean lethal dose (Do) and sensitive enhancement ratio (SER) were obtained with the colony formation assay after different treatments. The Do and SER of CMNa with radiotherapy groups are 1.483 and 1.683, the Do of radiotherapy only is 2.496. FITC and PI double labeled detect apoptosis to compare the difference between radiotherapy only and CMNa with different dose radiotherapy by flow cytometry. Apoptosis rates of CMNa with radiotherapy groups were significantly higher than radiotherapy group (P<0.01). The changes of cell homogeneous adhering capacity were observed by homotypic adhesion experiment. The homotypic adhesion experiments show that there are significant differences in the numbers of adhering cells among three groups after vibrating 40 and 60min(P<0.01).2. Evodiamine combined radioation inhibite proliferation, adhesion and migration of Tca-8113 cell line Cells growth inhibition was assessed by MTT after treated with indicated concentrations and time. Survival rates of Tca-8113 cells were decreased as increase of evodiamine concentration and time.The radiosensitization of evodiamine was measured by MTT assay. The cell survival rates of radiotherapy and evodiamien with radiotherapy groups were both decreased with the increase exposal doses and time. However, the cell survival rates of evodiamien with radiotherapy group was obviously lower than radiotherapy group on indicated exposal doses(2,4,6 Gy) and time(24,48,72 h) (P<0.01).The mean lethal dose (Do) and sensitive enhancement ratio (SER) were obtained with the colony formation assay after different treatments. The Do and SER of evodiamine with radiotherapy group were 1.84 and 1.35. Cells cycle was analyzed by PI labeled flow cytometry(FCM) after irradiation.Result indecates EVO with radiotherapy can blocked cells in G2/M phase. FITC and PI double labeled detect apoptosis to compare the difference between radiotherapy only and EVO with different dose radiotherapy by flow cytometry. Apoptosis rates of combination group were significantly higher than radiotherapy group (P<0.01). The changes of tumor cell homogeneous adhering and adhesion to endothelial cell capacity were observed by homotypic adhesion experiment and tumor cell adhesion to endothelial cell assay. Migration was evaluated by wound healing assay. There are significant differences in the numbers of adhering cells among three groups after vibrating 40 and 60min (P<0.01) in the homotypic adhesion experiments. The result of tumor cell adhesion to endothelial cell assay indicated that evodiamine with radiotherapy can inhibite Tca-8113 cells adhering to vascular endothelial cells. The mobility of evodiamine with radiotherapy groups was significantly lower than radiotherapy only and control (P<0.01). Expressions of caspase 3, caspase 9 and survivin were detected by real time PCR. EVO can significant increase expression of caspase 3 mRNA and caspase 9 mRNA, decrease expression of survivin mRNA.
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