低频超声联合微泡造影剂诱导人胆囊癌GBC-SD细胞凋亡的实验研究

目的探讨低频超声联合微泡造影剂对人胆囊癌GBC-SD细胞凋亡的影响。 方法将培养的人胆囊癌GBC-SD细胞分为4组，每组设5个复孔。其中空白对照组不进行任何处理；微泡组加200 μl的微泡造影剂混匀，使微泡浓度为20%，不进行超声辐照；低频超声组以超声辐照声强为0.45 W/cm²、超声脉冲波频率为1 MHz的低频超声连续波辐照，连续辐照30 s，不加造影剂；联合组加入200 μl的微泡造影剂，混匀后以1 MHz低频超声，连续波辐照30 s。各组以CCK-8法检测细胞增殖活性，并用流式细胞仪测定各组细胞的凋亡情况。 结果空白对照组、微泡组、低频超声组、联合组GBC-SD细胞增殖活性分别为0.9272±0.1173、1.0088±0.0628、0.2116±0.0101、0.1470±0.0029。联合组细胞增殖活性低于空白对照组、微泡组、低频超声组，差异均有统计学意义（t=14.846、30.637、13.661，P均<0.05）。各处理组间细胞凋亡率差异有统计学差异（F=2390.900，P<0.05），联合组细胞凋亡率为0.682±0.022，显著高于空白对照组的0.073±0.005、微泡组的0.031±0.003、低频超声组的0.259±0.012，差异均有统计学意义（P均<0.05）。 结论低频超声联合微泡造影剂能降低体外培养的人胆囊癌GBC-SD细胞增殖活性，明显诱导人胆囊癌GBC-SD细胞凋亡。

Apoptosis induced by low-intensity ultrasound combined with microbubbles in human gallbladder cancer GBC-SD cells

ObjectiveTo investigate the effect of low-intensity ultrasound combined with microbubbles on apoptosis of human gallbladder cancer GBC-SD cells. MethodsUltrasound with a frequency of 1 MHz and intensity of 0.45 W/cm² in continuous wave mode was used. Human gallbladder cancer GBC-SD cells were divided into four groups, including control group (A, without any treatment), microbubbles group (B, mixed with 20% microbubbles), ultrasound group (C, low frequency ultrasonic continuous wave irradiation for 30 s) and combined group (D, microbubbles combined with ultrasonic waves). Cell proliferation activity of each group was detected by CCK-8 assay, and apoptosis was detected by flow cytometry. ResultsProliferative activity of group A, B, C and D was 0.9272±0.1173, 1.0088±0.0628, 0.2116±0.0101 and 0.1470±0.0029, respectively. The proliferative activity of group D was lower than that of the other groups (t=14.846, 30.637 and 13.661, all P<0.05). Apoptosis was observed in all treated cells. Cell apoptosis rates of group D, A, B and C was 0.682±0.022, 0.073±0.005, 0.031±0.003 and 0.259±0.012, respectively. The apoptosis rate of group D was obviously higher than those of the other groups (F=2390.900, P<0.05). ConclusionLow-frequency ultrasound combined with microbubbles can reduce proliferative activity of human gallbladder cancer GBC-SD cells in vitro, and obviously induce the apoptosis of human gallbladder cancer GBC-SD cells.