正交设计法优化超声联合微泡诱导人前列腺癌细胞凋亡的研究

Optimization of ultrasound combined with microbubbles induced apoptosis in human prostate cancer cells according to the orthogonal design

目的采用正交实验设计,探讨超声联合微泡诱导人前列腺癌细胞的凋亡的最优参数组合。方法选定超声功率、辐照时间、微泡/细胞悬液体积比三个因素,又分别按超声功率15、20、25、30 mW/cm2,辐照时间30、60、90、120 s和微泡/细胞悬液体积比10%、20%、30%、40%,每因素四个水平设定正交设计表,按正交设计表进行超声辐照,辐照后细胞继续培养24 h,用流式细胞仪进行细胞凋亡检测。结果对细胞凋亡的影响因素,超声功率>微泡/细胞悬液体积>辐照时间;各因素水平影响:超声功率:15 mW/cm2>25 mW/cm2>20 mW/cm2>30 mW/cm2,辐照时间:30 s>60 s>120 s>90 s,微泡/细胞悬液体积:20%>10%>30%>40%。结论超声联合微泡诱导人前列腺癌细胞凋亡最佳组合为超声功率15 mW/cm2,辐照时间30 s,微泡/细胞悬液体积比20%。

Objective To investigate the optimal parameters of the ultrasonic combined with microbuble induced apoptosis in human prostate cancer cells according to the orthogonal design. Methods The ultrasonic power, irradiation time and microbubble/cell suspension volume were selected, then four levels of each factor were chosen, ultrasonic power （ 15 mW/cm2, 20 mW/cm2, 25 mW/cm2, 30 mW/cm2）, irradiation time （30 s, 60 s, 90 s, 120 s）, and micro-bubble / cell suspension volume （ 10%, 20%, 30%, 40% ）. Three-factor four-level orthogonal design table was set, and the human prostate cancer cells were radiated according to the orthogonal experimental design. After ultrasound radiation, the cells were cultured for 24 h. The apoptosis was detected by flow eytometry. Results The influence factors on the cell apoptosis： ultrasonic power 〉microbubble/cell suspension volume〉irradiation time; the influence of each factor level： uhrasonic power： 15 mW/cm2〉25 mW/cm2〉20 mW/cm2〉 30 mW/cm2; radiation time： 30 s〉60 s〉120 s〉90 s; microbubbles/eell suspension volume： 20%〉10%〉30%〉40%. Conclusion The optimal parameters of uhrasound combined with microbubble induced apoptosis in human prostate cancer cells are ultrasonic power of 15 mW/cm2, irradiation time of 30 s and micro-bubble/cell suspension volume ratio of 20%.