热疗通过提高活性caspase-3的表达促进食管癌EC109细胞凋亡
|
摘要
目的:探讨不同温度热疗诱导食管癌EC109细胞的生长抑制、细胞凋亡及其与天冬氨酸特异性半胱氨酸蛋白水解酶-3(caspase-3)蛋白表达变化的关系。方法:采用PCR仪精确控制温度,对食管癌EC109细胞行37℃(对照),43℃,45℃,47℃热疗处理,四甲基偶氮唑蓝(Tetrazolium bromide,MTT)法检测细胞生长抑制率,碘化丙啶(Propidium iodide,PI)/磷脂结合蛋白V(Annexin V)双染流式细胞仪测定细胞凋亡率,蛋白印迹(Western bolt)法检测caspase-3蛋白表达。结果:食管癌细胞在37℃,43℃,45℃处理下,细胞生长抑制率随温度升高而增多,并有显著性差异,43℃细胞凋亡率达到最高,三者差异显著。45℃和47℃生长抑制及凋亡率均无明显差异。总-caspase-3(pro-caspase-3)表达随温度升高而增多,而活性csapase-3(actived-caspase-3)在43℃表达最多。结论:热疗通过提高活性caspase-3表达诱导食管癌EC109细胞凋亡,在43℃凋亡率达到最高。
Objective:To explore the cell growth inhibition and apoptosis of esophageal cancer cells EC109 induced by hyperthermia in different temperature and its relation with the expression of caspase-3.Methods:Using PCR instrument to control the temperature precisely,cells treated at 37℃ as control,EC109 cells were treated at 43℃,45℃,47℃.The cell growth,apoptosis rate was detected by MTT assay and flow cytometry.The expression of caspase-3 was measured by Western Bolt.Results:The cell growth inhibition rates of EC109 cells were increased gradually at 37℃,43℃ and 45℃.The apoptosis rate at 43℃ was significantly higher than other groups.The cell growth inhibition rates and apoptosis rates had no significant difference between 45℃ and 43℃.Pro-Caspase-3 expression increased with increasing temperature,while actived-caspase-3 was expressed highest at 43℃.Conclusion:Hyperthermia induces apoptosis of EC109 cells by increasing the expression of activited-caspase-3 and the apoptosis rate reaches a peak at 43℃.
Objective:To explore the cell growth inhibition and apoptosis of esophageal cancer cells EC109 induced by hyperthermia in different temperature and its relation with the expression of caspase-3.Methods:Using PCR instrument to control the temperature precisely,cells treated at 37℃ as control,EC109 cells were treated at 43℃,45℃,47℃.The cell growth,apoptosis rate was detected by MTT assay and flow cytometry.The expression of caspase-3 was measured by Western Bolt.Results:The cell growth inhibition rates of EC109 cells were increased gradually at 37℃,43℃ and 45℃.The apoptosis rate at 43℃ was significantly higher than other groups.The cell growth inhibition rates and apoptosis rates had no significant difference between 45℃ and 43℃.Pro-Caspase-3 expression increased with increasing temperature,while actived-caspase-3 was expressed highest at 43℃.Conclusion:Hyperthermia induces apoptosis of EC109 cells by increasing the expression of activited-caspase-3 and the apoptosis rate reaches a peak at 43℃.
引文
[1]Parkin DM,Bray F,Ferlay J.Global cancer statistics,2002[J].CACancer J Clin,2005,55(2):74-108.
[2]中华人民共和国卫生部.全国第三次死因回顾抽样调查报告[M].北京:中国协和医科大学出版社,2008:10.
[3]詹宏杰.肿瘤热疗的研究[J].国外医学肿瘤学分册,2005,32:35-37.
[4]刘丽燕,王兴鹏,吴恺.胰腺癌SW1990细胞热疗温度及机制初探[J].胰腺病学,2005,5(3):143-145.
[5]Du C,Wang X.Smac,a mitochondrial protein that promotes cyto-chrome c-dependent caspase activation by eliminating IAP inhibi-tion[J].Cell,2000,102(1):33-42.
[6]Srinivasula SM,Hegde R,Saleh A,et al.A conserved XIAP-in-teraction motif in caspase-9 and Smac/DIABLO regulates caspaseactivity and apoptosis[J].Nature,2001,410:112-116.
[7]Chang HY,Yang X.Proteases for cell suicide:functions and regula-tion of caspases[J].Microbiol Mol Biol Rev,2000,64:821-846.
[8]Franckena M,Stalpers LJ,Koper PC,et al.Long-term improve-ment in treatment outcome after radiotherapy and hyperthermia inlocoregionally advanced cervix cancer:an update of the Dutch deephyperthermia trial Int[J].Radiat Oncol Biol Phys,2008,70(4):1176-1182.
[9]张明五,吴皓,吴逸明,等.不同温度下紫杉醇对人肺癌A549细胞抑制效应[J].中国公共卫生,2006,22(5):550.
[10]李晓红.放疗联合热疗治疗中晚期肺癌的临床疗效分析[J].临床肺科杂志,2008,13:501-502.
[11]黄培,周菊英.热疗在肿瘤治疗中的研究进展[J].现代肿瘤医学,2010,18(7):1460-1462.
[12]周梅菊.不同温度对人宫颈癌细胞凋亡和增殖的影响[A].中南大学博士学位论文[C].长沙:中南大学出版社,2009,77-78.
[13]Cheung TH,Chung TK,Lo KW,et al.Apoptosis-related pro-teins in cervical intraepithelial and squamous cell carcinoma of thecervix[J].Gynecol Oncol,2002,86:14-18.
[14]汪胜林,张振玉.Survivin与Caspase-3在光动力诱导肝癌细胞SMMC-7721凋亡中的作用[J].现代肿瘤医学,2010,18(7):1282-1284.
[15]Fukami T,Nakasu S,Baba K,et al.Hyperthermia induces translo-cation of apoptosis-inducing factor(AIF)an apoptosis in humanglioma cell lines[J].J Neurooncol,2004,70(3):319-331.
[2]中华人民共和国卫生部.全国第三次死因回顾抽样调查报告[M].北京:中国协和医科大学出版社,2008:10.
[3]詹宏杰.肿瘤热疗的研究[J].国外医学肿瘤学分册,2005,32:35-37.
[4]刘丽燕,王兴鹏,吴恺.胰腺癌SW1990细胞热疗温度及机制初探[J].胰腺病学,2005,5(3):143-145.
[5]Du C,Wang X.Smac,a mitochondrial protein that promotes cyto-chrome c-dependent caspase activation by eliminating IAP inhibi-tion[J].Cell,2000,102(1):33-42.
[6]Srinivasula SM,Hegde R,Saleh A,et al.A conserved XIAP-in-teraction motif in caspase-9 and Smac/DIABLO regulates caspaseactivity and apoptosis[J].Nature,2001,410:112-116.
[7]Chang HY,Yang X.Proteases for cell suicide:functions and regula-tion of caspases[J].Microbiol Mol Biol Rev,2000,64:821-846.
[8]Franckena M,Stalpers LJ,Koper PC,et al.Long-term improve-ment in treatment outcome after radiotherapy and hyperthermia inlocoregionally advanced cervix cancer:an update of the Dutch deephyperthermia trial Int[J].Radiat Oncol Biol Phys,2008,70(4):1176-1182.
[9]张明五,吴皓,吴逸明,等.不同温度下紫杉醇对人肺癌A549细胞抑制效应[J].中国公共卫生,2006,22(5):550.
[10]李晓红.放疗联合热疗治疗中晚期肺癌的临床疗效分析[J].临床肺科杂志,2008,13:501-502.
[11]黄培,周菊英.热疗在肿瘤治疗中的研究进展[J].现代肿瘤医学,2010,18(7):1460-1462.
[12]周梅菊.不同温度对人宫颈癌细胞凋亡和增殖的影响[A].中南大学博士学位论文[C].长沙:中南大学出版社,2009,77-78.
[13]Cheung TH,Chung TK,Lo KW,et al.Apoptosis-related pro-teins in cervical intraepithelial and squamous cell carcinoma of thecervix[J].Gynecol Oncol,2002,86:14-18.
[14]汪胜林,张振玉.Survivin与Caspase-3在光动力诱导肝癌细胞SMMC-7721凋亡中的作用[J].现代肿瘤医学,2010,18(7):1282-1284.
[15]Fukami T,Nakasu S,Baba K,et al.Hyperthermia induces translo-cation of apoptosis-inducing factor(AIF)an apoptosis in humanglioma cell lines[J].J Neurooncol,2004,70(3):319-331.