| Objective This study was designed to explore125I, hyperthermia,and combined use of the in vitro human tongue squamous cell carcinoma (of Tca8113) cells,human salivary adenoid cystic carcinoma (ACC-M) cell apoptosis,explore the best way that the use of125I radiotherapy combined with hyperthermia destruction of human tongue squamous cell carcinoma cells and human adenoid cystic carcinoma,and to provide a theoretical basis for the treatments of oral squamous cell carcinoma and adenoid cystic carcinoma.Methods Tca8113cells, ACC-M cells to5×105/ml density seeded in6-well plates,after receiving125I irradiation or/and43℃water bath until the adherent cells,cells were collected72hours later,for all testing.0.8mCi of125I radioactivity,placed in each well of six well plates at the center of the plate top and the bottom plate of each one of125I seeds. Experimental groups:blank control group (of Tca8113,the ACC-M-cells to normal culture),the experimental group one(125I+heat processing Tca8113cells);group two(125I processing Tca8113cells);group three(heating processing Tca8113cells);group four(125I+The heat treatment of ACC-M cells);group five(125I treatment of ACC-M cells);group six(heat treatment of ACC-M cells).Inverted microscope to observe the morphological changes of cells in each group.Flow cytometry, TUNEL method the percentage of apoptosis and cell cycle of cells in each group.Immunocytochemical staining of cells in each group of Bcl-2and Bax protein expression.All data were statistically analyzed using SPSS17.0statistical package,count data using the Wilcoxon rank sum test analysis and inspection.Results1.Under an inverted microscope to observe the control group,the125I group and heated group,125I+heated24hours the cells after treatment of the combined group morphological changes:After heat or/and125I treatment Tca8113cells rounded, wrinkled,slow growth,uneven distribution;ACC-M cells and nuclei smaller in size,loosely arranged cells were observed under inverted microscope cells float like.Put hyperthermia combination group and radiotherapy group and heated group compared to the more obvious changes in cell morphology.2.Flow cytometry showed that,Tca8113cells in each group72hours after treatment,compared with the control group,radiotherapy group,the heating group, put hyperthermia combined group apoptosis rate was increased,the difference was statistically significant.In addition to the radiotherapy group and heating group,p=0.513,the difference between each experimental group were statistically significant.From the cell cycle compared with the control group in each experimental group cells showed a significant G2/M phase arrest;ACC-M cells of each group compared with the control group72hours after treatment,the radiotherapy group,the heating group,put hyperthermia combined with apoptosis rate was significantly increased,the difference was statistically significant.From the cell cycle,compared with the control group,the cells of each experimental group showed a significant G2/M phase arrest;Between the two kinds of cells in each experimental group were compared,outside heated between groups p=0.817,the difference between the radiotherapy group,the thermal radiation group were statistically significant.3.1mmunocytochemical staining analysis of each group Tca8113cells and ACC-M cells handle72h after Bcl-2and Bax expression is:Tca8113cells and ACC-M cells after125I irradiation or/and heat treatment, Bax expression was enhanced,Bcl-2expression are weakened;put hyperthermia combined group and radiotherapy group and heated group compared to the change of Bax and Bcl-2expression is most significant. Conclusions1.125I and/or heating can induce the apoptosis of Tca8113and ACC-M cells,the combination of radiotherapy and hyperthermia synergistic sensitizing effect on apoptosis of these cancer cells.2.After72hours125I exposure and43℃heating80min,Tca8113and ACC-M cells cycle mainly for the G2/M phase arrest.3.125I radiotherapy with hyperthermia combined use of ACC-M cells apoptosis induced stronger than Tca8113cells. |
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