低强度超声对HeLa肿瘤细胞凋亡的诱导作用及其机制

目的探讨低强度超声在诱导HeLa肿瘤细胞凋亡中的作用及其机制。 方法将体外培养的HeLa肿瘤细胞根据超声干预强度分为阴性对照组（切断电源后给予假的超声干预）和0.5、1.3、2.0 W/cm²超声干预组，分别通过MTT实验测定细胞活力、存活率，HE染色检测细胞形态，细胞凋亡实验检测细胞凋亡率，流式细胞术测定ROS含量及Western blot实验检测caspase-12、survivin和内参蛋白β-actin的表达情况。多组间比较采用单因素方差分析，各个实验组与对照组比较采用Dunnet-t检验。 结果与阴性对照组比较，0.5、1.3、2.0 W/ cm²超声干预组的HeLa肿瘤细胞活力（99.23±1.56）﹪比（80.52±1.72）﹪、（54.31±1.69）﹪（27.75±1.26）﹪]、细胞存活率（99.91±1.51）﹪比（76.69±1.92）﹪、（52.57±1.63）﹪、（29.81±1.22）﹪]、survivin蛋白表达（51.19±0.21比43.46±0.34、25.28±0.29、18.32±0.3）均降低，ROS含量（11.21±0.45比24.34±1.23、38.26±2.47、52.18±1.56），细胞凋亡率（4.23±1.21）﹪比（24.16±1.91）﹪、（48.34±1.66）﹪、（70.27±0.98）﹪]、caspase-12蛋白表达（13.05±0.21比20.23±0.19、33.17±0.32、41.52±0.21）均升高，差异具有统计学意义（P均< 0.05）。 结论低强度超声可能通过诱导HeLa肿瘤细胞中caspase-12蛋白表达增加和survivin蛋白表达的减少而使HeLa肿瘤细胞发生凋亡。

Effects and mechanisms of low intensity ultrasound-induced apoptosis of HeLa tumor cells

objectiveTo investigate the role and mechanism of low intensity ultrasound induced apoptosis of HeLa tumor cells. MethodsAccording to the intensity of ultrasound intervention, HeLa tumor cells cultured in vitro were divided into blank control group (pseudo-ultrasound after power cut off) and setting up three ultrasound intervention groups (0.5, 1.3, 2.0 W/ cm², respectively) . Cell viability and survival rate were determined by MTT, and cell morphology was detected by HE staining. Apoptotic rate was detected by apoptosis experiment. ROS content was determined by flow cytometry and the expression of caspase-12, survivin and the internal reference protein β-actin was detected by Western blot. One-way ANOVA was used for comparison among multiple groups, and Dunnet-t test was used for multiple comparisons. Compared with blank control group, the cell viability (99.23﹪±1.56﹪vs 80.52﹪±1.72﹪, 54.31﹪±1.69﹪, 27.75﹪±1.26﹪, P < 0.05) , cell survival rates (99.91﹪±1.51﹪vs 76.69﹪±1.92﹪, 52.57﹪±1.63﹪, 29.81﹪±1.22﹪, P < 0.05) , ROS content (11.21±0.45 vs 24.34±1.23, 38.26±2.47, 52.18±1.56, P < 0.05) as well as the expression of surviving protein (51.19±0.21 vs 43.46±0.34, 25.28±0.29, 18.32±0.3, P < 0.05) were decreased in ultrasound intervention groups (0.5, 1.3, 2.0 W/cm², respectively) . While the apoptosis rate (4.23﹪±1.21﹪vs 24.16﹪±1.91﹪, 48.34﹪±1.66﹪, 70.27﹪±0.98﹪, P < 0.05) and the expression of caspase-12 protein (13.05±0.21 vs 20.23±0.19, 33.17±0.32, 41.52±0.21, P < 0.05) were increased. The difference was statistically significant (all P < 0.05) . ConclusionLow intensity ultrasound may induce the increase of caspase-12 protein expression and decrease of survivin protein expression in HeLa tumor cells, thus leading to apoptosis of HeLa tumor cells.