低频超声联合微泡诱导胰腺癌PANC-1细胞凋亡的线粒体机制  

Mitochondrial mechanism apoptosis induced by low-frequency ultrasound combined with microbubble agent in pancreatic carcinoma cell

作　　者：尚晓斌[1] 王子凡[1] 李卓[1] 魏林郁[2] 

机构地区：[1]新乡医学院第二附属医院功能检查科,453002 [2]新乡医学院生理学与神经生物学教研室,453002

出　　处：《实用医学杂志》2016年第23期3836-3840,共5页The Journal of Practical Medicine

基　　金：河南省教育厅科学技术研究重点项目(编号:14A310019)

摘　　要：目的:探讨低频超声联合微泡诱导人胰腺癌PANC-1细胞凋亡过程中线粒体凋亡通路关键分子表达和线粒体膜电位的改变。方法:对数生长期PANC-细胞分为空白对照组(A组)、单纯微泡组(B组)、单纯低频超声组(C组)和低频超声联合微泡组(D组);检测各组细胞凋亡形态、细胞凋亡率、线粒体膜电位和凋亡相关蛋白表达。结果:C组和D组出现绿染早期凋亡和红染中晚期凋亡PANC-1细胞;A、B、C和D组细胞凋亡率分别为(2.71±0.13)%、(3.69±0.18)%、(19.45±0.85)%和(33.36±1.12)%,D组凋亡率与余组比较差异均具有统计学意义(P<0.05);A、B、C和D组低Rho123荧光强度的细胞百分比分别为(3.63±0.34)%、(5.64±0.41)%、(21.78±0.48)%和(39.86±0.53)%,D组与余组比较差异均具有统计学意义(P<0.05);D组与其余各组比较,Bax表达上调、Bcl-2表达下调、CytC释放增加和活化Caspase-9蛋白表达增加。结论:低频超声联合微泡可以诱导人胰腺癌PANC-1细胞发生凋亡,其机制可能与线粒体凋亡通路关键信号分子Bcl-2、Bax、CytC、Caspase-9表达改变和线粒体膜电位△Ψ下降等密切相关。Objective To study whether low-frequency ultrasound combined with microbubble agent induced apoptosis in pancreatic carcinomaPANC-1 cell via mitochondrial pathway. Methods Cells were divided into blank group（A）, microbubble agent group（B）, low-frequency ultrasound group（C） and low frequency ultrasound combined with microbubble agent group （D）. Phase contrast microscope was used to observe the cells morphology changed; TUNEL staining flow Cytometry method was used to observe the apoptosis; The mitochondria transmembrane potential （AXiOm） change was calculated by Rho123fluorescence probe; The expression changes of apoptosis-related protein in mitochondrial pathwaywere detected by Western Blot. Results Annexin V-FITC/ PI double dye fluorescence microscope observed in group C and group D appear green dye early apoptosis cells and red dye middle-late apoptotic cells; Group D apoptosis rate is higher than the rest of the group and the A t^m value is lower than the rest of the group, the differences were statistically significant respectively; Group D compared to the rest of the group, the expression of Bax raised, Bel-2 expressiondecreased, CytC release increased and activated Caspase-9 protein expressionraised. Conclusion Low frequency ultrasound combined with microbubble agent induced the PANC-1 cells apoptosis may via the mitochondrial-dependent apoptosis pathway.

关 键 词：低频超声 微泡剂 细胞凋亡 线粒体凋亡通路 

分 类 号：R735.9[医药卫生—肿瘤]