| The most common of oral squamous cell carcinoma (OSCC) is tongue squamous～cell carcinoma (TSCC),which is fast-growing, strong soaking, high malignant intensity and often occurs cervical lymph node in early stage. TSCC have the most significant proportion among malignant oral and maxillofacial tumors, reaching about one third in China. Now,the therapy of TSCC mainly relies on combined and sequential therapy which surgical treatment is dominant.But there are others hard problem,for example local recurrence easily,distant metastasis,if the surgery only selected. Therefore, For TSCC, the therapy of postoperation should be combinated with low side effects of chemotherapy drugs, reducing the recurrence rate and improving survival. So, the research of postoperative chemotherapy is essential for malignant tumor.Apigenin, a natural flavonoid,has been shown to be low toxicity,non-mutagenic and high efficacy,and exist in many kinds of fruits and vegetables extensively. Domestic and international study confirmed that apigenin can inhibit occurrenceand development of tumors, such as gastrointestinal tumors, urinary tumors, germina tumors and so on. A large number of experiments had proved that the role of apigenin is growing concern in the treatment of malignant tumors.Objective:1. To investigate the inhabition of apigenin on proliferation of Tca8113 cells.2. To explore the possible mechanism of inducing apoptosis of tumor cells and affected proliferation of apigenin.3. To research the effect of the therapy which apigenin combine hyperthermia on tumor cells,and provide theoretical basis for theraping TSCC. Materials and Methods:Culture Tca8113 cells and act on the cell with different concentration of apigenin.After certain time, observing the growth states and morphological changes of the tongue cancer cell by invert microscope. Measuring the effects on inhibitory rate of tongue cancer Tca8113 cells cell growth by using MTT. Observing the medicine's effects on change of cell shape and apoptosis of cells by acridine orange staining. Checking the apoptosis utilizing the Annexin V-FITC/PI double staining. Detecting the cell cycle process utilizing the flow cytometry technique. Adopting SPSS 13.0 software for related statistical analysis, using the chi-square test,chi-square test division,repeated measurement of variance.Took the significance testing level a=0.05.Results:1. Apigenin inhibition of Tca8113 cells'growth obviously:after different time, different concentrations of apigenin were all effective to inhibit the Tca8113 cells; with the higher concentration of apigenin and the extension of time,the strength of inhibiton to be enhanced.2. Observation with inverted microscopy:after dealt with 40μmol/L for 48 hours, cell size become smaller and scattered with spindle-shaped cells and state clearly; and the capacity of adhesion diminished.3. With acridine orange staining:Being treated by apigenin,the nucleus of cells was yellow green and concentrated inside the karyotheca, and the cell membrane was expanded in bubble form.4. In result of flow cytometry after dealt with apigenin:G1-phase cells were increasing but S-phase cells were decreasing; the cells stopped at G1/S phase, and the difference shown was significant in statistical terms compared with the control group (P<0.05). 5. Cell apoptosis tested with Propidium lodide-Annexin V-FITC double staining and flow cytometry:after 24 hours' action of 40μmol/L and 80μmol/L apigenin: the cell apoptosis rate was increasing and correlation with dosage-effect and time-effect(the higher the concentration and the longer the action time, the higher was the apoptosis rate). There was an significant in statistical terms compared with the control group(P<0.05).6. With the increase of temperature and concentration, the tumor cell apoptosis rate increased, the difference shown was significant in statistical terms.(P<0.05).Conclusions:1. The growth of Tca8113 cells is inhibited by Apigenin,whose intensity is positive correlection with the concentration and action time.2. Apigenin is able to induce the apoptosis of Tca8113 cells; the cell apoptosis rate was increasing with the action time and dosage increase.3. The cell cycle of Tca8113 cells was influenced by apigenin,and the cells stopped at Gl/S phase. This could also affect the apigenin through the means of the occurrence and development of tumors.4. Apigenin may be synergistic combination of hyperthermia on Tca8113 cells and promote their apoptosis. |
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