秦娟;宋国林;王建三;林虹;刘卿;陈锦云;王嫣;
QIN Juan;SONG Guolin;WANG Jiansan;LIN Hong;LIU Qin;CHEN Jinyun;WANG Yan;Department of Gynaecology, Guiyang Maternal and Child Health Hospital;Department of ICU, 2nd Affiliated Hospital of Guizhou University of Traditional Chinese Medicine;State Key Laboratory of Ultrasound Engineering in Medicine Co-Founded by Chongqing and the Ministry of Science and Technology, College of Biomedical Engineering, Chongqing Key Laboratory of Biomedical Engineering,Chongqing Collaborative Innovation Center for Mi

• 1:贵阳市妇幼保健医院妇科
• 2:贵州省中医药大学第二附属医院重症医学科
• 3:重庆医科大学生物医学工程学院省部共建国家重点实验室培育基地-重庆市超声医学工程重点实验室重庆市生物医学工程学重点实验室

摘要(Abstract)：

目的:探讨低强度超声辐照对人宫颈癌HeLa细胞凋亡及半胱氨酸天冬氨酸特异性蛋白酶-12(Casepase-12)蛋白表达的影响。方法:采用低强度超声辐照人子宫颈癌HeLa细胞株,于辐照后6及24 h时,Annexin V/PI双染法检测HeLa细胞的凋亡率,透射电子显微镜观察细胞的结构变化,Western blot法测定细胞匀浆中Caspase-12蛋白表达水平,免疫组织化学法检测Caspase-12蛋白在HeLa细胞中的阳性表达率。结果:低强度超声辐照后6 h时HeLa细胞的凋亡率、细胞Caspase-12蛋白表达水平及Caspase-12蛋白在HeLa细胞中的阳性表达率显著高于对照组,差异有统计学意义(P<0.05);辐照后24 h时变化更显著,差异有统计学意义(P<0.01);透射电子显微镜下可见,辐照后6 h时HeLa细胞内开始出现凋亡小体,24 h时胞浆浓缩、内质网变疏松并与胞膜融合、形成空泡;辐照后6 h及24 h时,Caspase-12蛋白在HeLa细胞胞浆中表达、呈棕黄色颗粒,但在对照组细胞中无表达。结论:低强度超声辐照可以诱导人宫颈癌HeLa细胞凋亡,并随着辐照后时间延长而增加,其机制可能与相关凋亡蛋白Caspase-12表达水平上调有关。
Objective: To study the effect of ultrasonic irradiation with low intensity on the protein expressions of Caspase 12 and apoptosis of HeLa cells. Methods: Human cervical cancer HeLa cells were cultured with ultrasonic irradiation of low intensity. Flow cytometry was used to detect the apoptosis rate in 6 and 24 hours after ultrasound irradiation. The structural changes of cells under ultrasound irradiation were observed by scanning electron microscope. The protein expression level of Caspase 12 in the cervical cancer cells was determined. Results: The apoptosis rate of HeLa cells 6 hours after ultrasound irradiation was significantly higher than that of the control group(P<0.05). The apoptosis rate increased in 24 hours after surgery, significantly higher than that of the control group(P<0.01). Apoptosis body in HeLa cells was induced in 6 hours after ultrasound irradiation. The expression of related apoptosis protein Caspase 12 was activated after ultrasound irradiation, compared with the control group(P<0.05). The expression of Caspase 12 in 24 hours after irradiation increased significantly compared with the control group(P<0.01). Conclusion: Low intensity ultrasound can induce apoptosis of HeLa cells and the apoptosis rate increases with the time of irradiation. Caspase 12 can be activated under ultrasound irradiation, and the expression of Caspase 12 increases with time of irradiation.

关键词(KeyWords)： 超声多普勒,低强度;宫颈癌;HeLa细胞;细胞凋亡;凋亡蛋白;半胱氨酸天冬氨酸特异性蛋白酶-12;免疫组织化学法
ultrasound,low intensity;cervical cancer;HeLa cells;apoptosis;apoptosis protein;Caspase 12;immunohistochemistry

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81560287);; 贵州省科技厅基金[黔科合J字(2013)2020]

作者(Authors): 秦娟;宋国林;王建三;林虹;刘卿;陈锦云;王嫣;
QIN Juan;SONG Guolin;WANG Jiansan;LIN Hong;LIU Qin;CHEN Jinyun;WANG Yan;Department of Gynaecology, Guiyang Maternal and Child Health Hospital;Department of ICU, 2nd Affiliated Hospital of Guizhou University of Traditional Chinese Medicine;State Key Laboratory of Ultrasound Engineering in Medicine Co-Founded by Chongqing and the Ministry of Science and Technology, College of Biomedical Engineering, Chongqing Key Laboratory of Biomedical Engineering,Chongqing Collaborative Innovation Center for Mi

DOI: 10.19367/j.cnki.1000-2707.2019.10.009

参考文献(References)：

• [1] SCHOLZ C,WEISSENBACHER T,FRIESE K,et al.Cervical cancer-minimally invasive treatment[J].MMW Fortschr Med,2010,152(43):36-37.
• [2] KROKIDIS M E,KITROU P,SPILIOPOULOS S,et al.Image-guided minimally invasive treatment for small renal cell carcinoma[J].Insights Imaging,2018,9(3):385-390.
• [3] FERIL L B,KONDO T,CUI Z G,et al.Apoptosis induced by the sonomechanical effects of low intensity pulsed ultrasound in a human leukemia cell line[J].Cancer Lett,2005,221(2):145-152.
• [4]付贞花,汪炼,凌玲,等.宫颈上皮内瘤样病变Ⅰ级经聚焦超声治疗前后局部P16、Ki-67基因及Fas表达的变化[J].中国介入影像与治疗学,2016,(4):230-233.
• [5]NING Z,ZHU Z,WANG H,et al.High-intensity focused ultrasound enhances the effect of bufalin by inducing apoptosis in pancreatic cancer cells[J].Onco Targets Ther,2019,12(12):1161-1170.
• [6] GUO X,GUO S,PAN L,et al.Anti-microRNA-21/221 and microRNA-199a transfected by ultrasound microbubbles induces the apoptosis of human hepatoma HepG2 cells[J].Oncol Lett,2017,13(5):3669-3675.
• [7] SALIEV T,FERIL L B,OGAWA K,et al.Induction of apoptosis in U937 cells by using a combination of bortezomib and low-intensity ultrasound[J].Med Sci Monit,2016,22(22):5049-5057.
• [8] POFF J A,ALLEN C T,TRAUGHBER B,et al.Pulsed high-intensity focused ultrasound enhances apoptosis and growth inhibition of squamous cell carcinoma xenografts with proteasome inhibitor bortezomib [J].Radiology,2008,248(2):485-491.
• [9] 王萍,夏新蜀,王志刚.低强度超声壤联合光敏剂对癌细胞的杀伤作用[J].激光杂志,2009,30(4):75-76.
• [10]LI H X,ZHENG J H,JI L,et al.Effects of low-intensity ultrasound combined with low-dose carboplatin in an orthotopic hamster model of tongue cancer:A preclinical study[J].Oncol Rep,2018,2(13):1-9.
• [11]YU T,BAI J,HU K,et al.The effect of free radical scavenger and antioxidant on the increase in intracellular adriamycin accumulation induced by ultrasound[J].Ultrason Sonochem,2003,10(1):33-35.
• [12]杨志宏,王智彪,胡凯,等.聚焦超声对卵巢癌细胞株COC1细胞凋亡和Bcl-2/bax基因表达的影响[J].中国医学物理学杂志,2004,21(4):207-210.
• [13]吴晓辉,陶裕川,汪峰,等.低强度超声联合微泡的声空化效应介导大鼠C_6胶质瘤细胞凋亡的研究[J].华中科技大学学报(医学版),2014,43(1):6-11.
• [14]张震,戚勋,徐克,等.低强度超声逆转胶质瘤ABCG2表达的实验研究[J].中国医科大学学报,2013,42(6):488-492.
• [15]邱日想,王燕莉,陈洋娣,等.低强度超声联合托泊替康诱导人肝癌细胞凋亡的研究[J].中国临床药理学与治疗学,2012,17(12):1384-1388.
• [16]崔海东,吕岩红,曹斌,等.低强度超声联合原卟啉IX诱导人舌鳞癌SAS细胞凋亡作用的研究[J].口腔医学研究,2011,27(7):580-582.
• [17]林艳端,申锷,白文坤,等.低频低功率超声联合微泡对DU145细胞及PC3细胞早期凋亡的影响[J].临床超声医学杂志,2014,16(1):1-4.
• [18]曾宇鹏,谢席胜.高尿酸对肾小管上皮细胞HK-2增殖、凋亡的影响及其机制研究[J].实用医学杂志,2018,34(24):4042-4045.
• [19]何金玲,燕子,曾先燕,等.机械创伤升高大鼠心肌细胞凋亡蛋白表达[J].基础医学与临床,2019,39(1):16-21.
• [20]牛陵川,邹文兵,张炼,等.乳腺癌高强度聚焦超声消融后残留肿瘤增殖能力的变化[J].上海交通大学学报(医学版),2010,30(2):200-203;207.
• [21]WILLIAM D,O'BRIEN J R.Ultrasound-biophysics mechanisms[J].Prog Biophys Mol Bio,2007,93(1-3):212-255.
• [22]FENG Y,TIAN Z,WAN M.Bio-effects of low intensity ultrasound in vitro:apoptosis,protein profile alteration and potential molecular mechanism [J].J Ultrasound Med,2010,29(6):963-974.
• [23]YU T,BAI J,HUK,et al.The effect of free radical scavenger and antioxidant on the increase in intracellular adriamyc in accumulation induced by ultrasound [J].Ultrasond Sonochem,2003,10(1):33-35.
• [24]YU T,HU K,BAI J,et al.Reversal of adriamycin resistance in ovarian carcinoma cell line by combination of verapamil and low-level ultrasound [J].Ultrason Sonochem,2003,10(1):37-40.
• [25]崔海东,吕岩红,曹斌,等.低强度超声联合原卟啉IX诱导人舌鳞癌SAS细胞凋亡作用的研究[J].口腔医学研究,2011,27(7):580-582.

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