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生长抑制因子诱导膀胱肿瘤细胞凋亡的研究
引用本文:Li L,He DL,He H,Wang XY,Zhang LL,Luo Y,Nan XY. 生长抑制因子诱导膀胱肿瘤细胞凋亡的研究[J]. 中华医学杂志, 2005, 85(25): 1766-1769
作者姓名:Li L  He DL  He H  Wang XY  Zhang LL  Luo Y  Nan XY
作者单位:710061,西安交通大学医学院第一附属医院泌尿外科
基金项目:国家自然科学基金资助项目(30170937)
摘    要:目的探讨生长抑制因子(PML)诱导膀胱肿瘤细胞凋亡的分子机制。方法应用脂质体Lipofectamine2000分别将重组可诱导表达的真核表达载体PMEP4/PML和对照PMEP4空载体转染到膀胱肿瘤细胞系UM-UC-2中,300μg/ml潮霉素B筛选稳定表达PML的抗性克隆。激光共聚焦检测PML蛋白表达。DNAladder法检测肿瘤细胞凋亡。Western印迹法检测凋亡相关蛋白表达情况。结果经5μmol/L的CdSO4诱导表达后,激光共聚焦显微镜观察发现转染PMEP4/PML组的细胞核内有散在的斑点样的黄绿色荧光亮点,而转染空载体PMEP4组细胞未见特异性的荧光斑点表达。DNA梯度法显示转染PML细胞在诱导PML表达后24h出现梯状凋亡条带。Western印迹法检测凋亡相关蛋白半胱氨蛋白水解酶3、活化PARP上调,而survivin的表达水平下调。结论PML诱导膀胱肿瘤细胞凋亡可能与上调半胱氨蛋白水解酶3、活化PARP蛋白,抑制survivin的表达有关。

关 键 词:肿瘤细胞凋亡 生长抑制因子 Western印迹法 膀胱 半胱氨蛋白水解酶 survivin 凋亡相关蛋白 PML蛋白表达 激光共聚焦 真核表达载体 诱导表达 肿瘤细胞系 mol/L 分子机制 方法应用 抗性克隆 稳定表达 表达情况 细胞核内

The molecular mechanism of PML in inducing apoptosis of bladder cancer cell line
Li Lei,He Da-lin,He Hui,Wang Xin-yang,Zhang Lin-lin,Luo Yong,Nan Xun-yi. The molecular mechanism of PML in inducing apoptosis of bladder cancer cell line[J]. Zhonghua yi xue za zhi, 2005, 85(25): 1766-1769
Authors:Li Lei  He Da-lin  He Hui  Wang Xin-yang  Zhang Lin-lin  Luo Yong  Nan Xun-yi
Affiliation:Institute of Urology, No.1 Hospital, Xi'an Jiaotong University, Shanxi 710061, China.
Abstract:OBJECTIVE: Study the affection of inducing expression of PML in the apoptosis and the molecular mechanism of bladder cancer. METHODS: PMEP4/PML inducible expression vector was transfected into bladder cancer UM-UC-2 cells by lipofectamine 2000 system. The positive clone cells were selected by 300 microg/ml hygromycin B and confirmed by laser confocal imaging system. Then, using the in vitro DNA ladder apoptotic assay and Western blot, the affection of inducing expression of PML on apoptosis and its molecular mechanism of bladder cancer cell was studied. RESULTS: Comparing with the vector control group, PML specific nuclear speckle significantly increased in the PMEP4/PML bladder cells. DNA ladder assay demonstrated bladder cancer cell expressing PML occurred apoptosis while the control vector cells were not influenced. Overexpression of PML could reduce Survivin expression and upregulate caspase3 and cleaved PARP protein expression CONCLUSION: Our results demonstrated that overexpression of PML could induce bladder cancer cell apoptosis through the caspase dependent pathways.
Keywords:Bladder cancer  Growth inhibitors  Promyelocytic Leukemia   gene  Apoptosis
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