20 kHz超声联合微泡诱导人前列腺癌PC3细胞的细胞自噬及早期凋亡

目的探讨低频、低功率超声联合微泡造影剂诱导人雄激素非依赖性前列腺癌PC3细胞早期细胞凋亡及自噬。方法以频率20kHz、声功率80mW超声连续波辐照人前列腺癌PC3细胞悬液60s,之后分为单纯微泡组(A)、单纯超声组(B)、超声联合微泡组(C)、空白对照组(D),分别给予相应处理。辐照后继续培养24h,以流式细胞仪检测细胞早期凋亡情况,吖啶橙染色荧光显微镜观察细胞胞浆内酸性囊泡,透射电镜观察细胞自噬泡。结果 4组细胞早期凋亡率差异有统计学意义(P0.01);两两比较,除A组与D组差异无统计学意义(P0.05)外,余差异均有统计学意义(P0.01)。C组细胞核基本正常,胞浆内可见大量发红色荧光的酸性囊泡及大量由双层膜包裹的自噬泡或自噬体。D组细胞形态基本正常,胞浆内未见到明显自噬泡形成。结论低频低功率超声联合微泡造影剂能明显提高人雄激素非依赖型前列腺癌PC3细胞的早期凋亡率,并可促进细胞自噬。

Apoptosis and autophagy induced by 20 kHz ultrasound combined with microbubbles in human prostate cancer PC3 cells

Objective To investigate the early apoptosis and autophagy induced by low-frequency and low-power ultrasound combined with microbubbles in PC3 cells. Methods Ultrasound with frequency of 20 kHz and intensity of 80 mW in continuous wave mode was used. PC3 cells were divided into four groups, i.e. microbubbles group (group A), ultrasound group (group B), ultrasound combined with microbubbles group (group C) and control group (D). Twenty-four h after irradiation, the early apoptosis was detected using flow cytometry, the acid vesicular organelles were detected by acridine orange fluorescence staining, and transmission electron microscopy was used to observe autophagosomes. Results The early apoptosis rate among 4 groups were statistically different (P<0.01). Multiple comparison showed no statistical differences except the data between group A and D. Acridine orange fluorescence staining showed formation of acid vesicular organelles in the cytoplasm with normal nucleus and lots of autophagosomes with double-membrane structure were detected in group C, while in group D cells were basically normal. Conclusion Low-frequency and low-power ultrasound combined with microbubbles can induce apoptosis and autophay in human prostate cancer cells PC3 cells.