目的探讨亚高温热化疗的体外抗肿瘤作用。方法(1)采用MTT法(四甲基偶氮唑盐比色法)观察紫杉醇对人乳腺癌细胞(MCF-7)生长的影响,筛选出有效实验浓度。(2)将体外培养的MCF-7细胞分为空白对照组、化疗组(紫杉醇)、热疗组(聚束微波加热)及热化疗组(聚束微波加热联合紫杉醇),各组MCF-7细胞分别经相应干预后,采用流式细胞术(Annexin V-FITC/PI)检测上述各组MCF-7细胞凋亡情况。结果(1)热疗后48 h,41℃热疗组细胞凋亡率高于空白对照组、40℃、40.5℃热疗组及热疗后24 h组(P<0.05)。(2)5 μg/mL化疗干预48 h、10 μg/mL化疗干预24 h细胞凋亡率均分别高于其他各组(P<0.05)。(3)5 μg/mL热化疗组干预24 h、48 h后细胞凋亡率均随着热疗温度的升高而升高(P<0.05);10 μg/mL热化疗组干预24 h后细胞凋亡率随着热疗温度的升高而升高,干预48 h后细胞凋亡率随着热疗温度的升高而降低。(4)细胞凋亡率的顺序为:5 μg/mL热化疗组48 h>10 μg/mL热化疗组24 h>10 μg/mL化疗组24 h>5 μg/mL化疗组48 h>41℃热疗组>空白对照组。结论在体外,一定温度的热疗可杀伤肿瘤细胞,热疗联合化疗协同杀伤肿瘤细胞作用更明显,但并非化疗药物浓度越高越有效,温度越高越有效。
ObjectiveTo investigate the effect of thermochemotherapy on anti-tumor in vitro. Methods(1) The effective concentration of paclitaxel (PTX) on proliferation of human breast cancer cell line (MCF-7) was determined by using MTT assays. (2) MCF-7 cells cultured in vitro were divided into control group, chemotherapy group(PTX), hyperthermia group(microwave) and thermochemotherapy group (microwave and PTX), cells in each group recived corresponding intervention, flow cytometry was used to detect the cell apoptosis. Results(1) After 48 hours of thermotherapy, the cell apoptosis rate in 41℃ thermotherapy group was significantly higher than that in the control group, 40℃thermotherapy group, 40.5℃ thermotherapy group, and also significantly higher that in the thermotherapy for 24 hours group. (2) Chemotherapy for 48 hours in 5μg/mLPTX and chemotherapy for 24 hours in 10 μg/mL PTX got the highest cell apoptosis rate compared to the other groups. (3) Intervention for 24 hours and 48hours, the cell apoptosis rate in 5 μg/mL PTX thermochemotherapy group were both increased with increasing temperature , but in 10 μg/mL PTX thermochemotherapy group, intervention for 24 hours, the cell apoptosis rate was increased with increasing temperature ,while decreases with the rise of the temperature after treating for 48 hours. (4) The sequence of cell apoptosis rate was 5 μg/mL PTX thermochemotherapy group intervention for 48 hours >10 μg/mL PTX thermochemotherapy group intervention for 24 hours >10 μg/mL PTX intervention for 24 hours >5 μg/mL PTX intervention for 48 hours >41℃ thermotherapy group >control group. Compare with chemotherapy group and hyperthermia group, thermochemotherapy group(40.5℃) could induce more MCF-7 cells apoptosis, 41℃/(5 μg/ml) thermochemotherapy had more apoptosis cells than 40.5℃thermochemotherapy group. However, MCF-7 cells proliferation enhanced in 40℃/(5 μg/ml)thermochemotherapy group. ConclusionsIn vitro, certain temperature can kill tumor cells, hyperthermia therapy has synergistic effect with chemotherapy, however, it is not that the higher concentration of chemotherapy drugs, the more anti-tumor satisfaction is, so does the temperature.