热疗诱导肺癌细胞凋亡机制的实验研究.docVIP

热疗诱导肺癌细胞凋亡机制的实验研究 蒋东1 李芳2 葛锦峰1 郑世营1 * （1．苏州大学附属第一医院心胸外科，江苏 苏州 215006； 2．苏州大学基础医学与生命科学学院 人体解剖与组织胚胎系 江苏 苏州 215006） 基金项目：江苏省社会发展重大课题资助项目（BS2005047） 通讯作者：郑世营* 摘要 目的:观察热疗对人肺癌A549、H1299细胞生长与凋亡的影响并探讨其机制。方法:采用四甲基偶氮唑蓝(MTT)比色法检测热疗对人肺癌A549、H1299细胞生长的抑制作用；采用流式细胞技术观察热疗对人肺癌A549、H1299细胞周期和凋亡的影响；采用RT-PCR检测凋亡相关基因P53及Casepase-3的表达。结果：热疗对人肺癌A549、H1299两种细胞的生长均有抑制作用。热疗可以明显改变人肺癌A549细胞生长周期的分布，G0/G1期细胞明显减少，但对人肺癌H1299细胞生长周期的分布影响不大。热疗A549细胞P53蛋白的表达明显增强，不能诱导H1299细胞P53蛋白的表达The Research of the Effect of Thermotherapy on Mechanism of Apoptosis in Lung Cancer Cells Abstract: Objective: To observe the effects of thermotherapy on the growth, apoptosis of Human Lung Cancer cells of A549, H1299, and to investigate its possible mechanism, under different temperature. Methods: The inhibition ability of thermotherapy on A549 cells and H1299 cells were determined by MTT, the apoptosis and cell cycle of A549 cells and H1299 cells were analyzed by FACS, and RT-PCT was adopted to detect the expression of the apoptosis related gene of p53 and Caspase-3. Results: The thermotherapy did have inhibitory effect both on the growth of A549 cells and H1299 cells, and it could change the cell cycle of A549 cells, and reduce its cellular proportion in G0/G1 phase, significantly. On the contrary the cell cycle of H1299 remained same before and after the thermotherapy. In addition, after thermotherapy the expression of P53 in A549 cells but not in H1299 was found increased, while the expression of Caspase-3 increased in both A549 and H1299 cells, and especially in A549 cells. Conclusions: The thermotherapy can prevent the growth, affect the growth circle and induce the apoptosis of lung cancer cells through P53. No matter under which heating temperature the inhibitory rate of cell growth and the rate of apoptosis in A549 cells was always high than it was in H1299 cells. 近年来肿瘤热疗已成为继手术、化疗、放疗、生物治疗之后的对肿瘤的第五类有效治疗手段。肺癌目前的发病率和死亡率在我国男性中均较高,约65%的肺癌患者确诊时已属晚期,失去手术机会，因而包括热疗在内的综合治疗在晚期肺癌的治疗中占有重要的地位。本实验以我们培养的A549、H1299细胞为实验对象,主要观察不