根皮素诱导前列腺癌PC-3细胞凋亡并抑制其裸鼠移植瘤的生长

发布时间：2018-06-16 12:33

本文选题：前列腺肿瘤 + 根皮素　；参考：《肿瘤》2017年06期

【摘要】：目的 :探讨根皮素对前列腺癌PC-3细胞凋亡和周期的影响,及对其裸鼠移植瘤生长的影响。方法 :不同浓度的根皮素(0、20、50和100μmol/L)分别作用于人正常前列腺基质细胞WPMY-1和前列腺癌PC-3和DU145细胞24 h后,倒置光学显微镜下观察细胞形态的变化,并采用CCK-8法检测各组细胞活性的变化。用不同浓度的根皮素(0、20、50和100μmol/L)处理PC-3细胞24 h后,采用DAPI染色法观察细胞核破裂的情况,FCM法检测根皮素处理后前列腺癌PC-3细胞凋亡率和细胞周期分布的变化;同时,采用蛋白质印迹法检测细胞凋亡相关蛋白剪切型聚腺苷二磷酸核糖聚合酶1(cleaved-poly ADP-ribose polymerase-1,cleaved-PARP-1)、cleaved-caspase 3、cleaved-caspase 8、cleaved-caspase 9、X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)、survivin、Bcl-2、Bax以及细胞周期蛋白cyclin B1表达水平的变化。建立人前列腺癌PC-3细胞裸鼠皮下移植瘤模型,观察根皮素腹腔注射治疗后小鼠移植瘤生长体积以及体质量的变化,并计算抑瘤率。采用蛋白质印迹法检测小鼠肿瘤组织中凋亡相关蛋白的表达水平。结果:根皮素浓度为50和100μmol/L时,对人正常前列腺基质细胞WPMY-1的形态没有影响,仅对前列腺癌PC-3和DU145细胞的形态有影响。与空白对照组相比,根皮素浓度为20、50和100μmol/L时对前列腺癌PC-3和DU145细胞活力均有明显的抑制作用(P值均0.01)。采用浓度为20、50和100μmol/L的根皮素处理PC-3细胞24 h后,PC-3细胞的核破裂率分别为(27.0±2.0)%、(47.0±1.5)%和(63.0±3.0)%,明显高于空白对照组的(3.0±1.0)%(P值均0.001);凋亡率分别为(20.0±1.0)%、(32.0±2.0)%和(42.6±1.5)%,明显高于空白对照组的(4.0±1.0)%(P值均0.001)。根皮素(100μmol/L)处理PC-3细胞24 h后,细胞中cleaved-PARP-1、cleaved-caspase 3、cleaved-caspase 8和cleaved-caspase 9蛋白的表达水平均较空白对照组明显升高(P值均0.01),而XIAP、survivin和Bcl-2蛋白的表达水平均明显降低(P值均0.001)。PC-3细胞被阻滞在G2/M期,并且cyclin B1蛋白的表达水平明显降低(P0.001)。根皮素可以抑制裸鼠移植瘤的生长(P0.05),并上调肿瘤组织中cleaved-PARP-1和cleaved-caspase 3蛋白的表达水平,下调survivin蛋白的表达水平(P值均0.001),但是对小鼠体质量无明显影响(P0.05)。结论 :根皮素通过下调cyclin B1、XIAP、survivin和Bcl-2蛋白水平,以及上调cleaved-PARP-1、cleaved-caspase 3、cleaved-caspase 8和cleaved-caspase 9蛋白的表达水平,诱导PC-3细胞周期阻滞和细胞凋亡,最终抑制肿瘤细胞生长。
[Abstract]:Aim: to investigate the effects of hirsutin on apoptosis and cell cycle of prostate cancer PC-3 cells and the growth of xenografts in nude mice. Methods: human normal prostate stromal cells (WPMY-1) and prostate cancer PC-3 and DU145 cells were treated with different concentrations of rotadermatine (50 渭 mol / L) and 100 渭 mol / L (100 渭 mol / L) for 24 h, respectively. The morphological changes of the cells were observed under inverted optical microscope. The changes of cell activity in each group were detected by CCK-8 method. PC-3 cells were treated with different concentrations of xanthoxylin 20 and 100 渭 mol / L for 24 h. The nuclear breakdown was observed by DAPI staining and the apoptosis rate and cell cycle distribution of PC-3 cells were detected by FCM. The expression of 1(cleaved-poly ADP-ribose Polymerase 1(cleaved-poly ADP-ribose Polymerase (1(cleaved-poly ADP-ribose Polymerase 1) cleaved-caspase 3 cleaved-caspase 8 cleaved-caspase 9 X linked apoptosis inhibitor protein X-linked inhibitor of apoptosis protein XIAPP survivin Bcl-2 Bcl-2 Bax and cell cycle protein cyclin B1 were detected by Western blot. The subcutaneous transplanted tumor model of human prostate cancer PC-3 cells was established in nude mice. The growth volume and body mass of transplanted tumor were observed after intraperitoneal injection of keratin, and the tumor inhibition rate was calculated. The expression of apoptosis-related proteins in mouse tumor tissues was detected by Western blot. Results: at concentrations of 50 渭 mol / L and 100 渭 mol / L, there was no effect on the morphology of human normal prostate stromal cells WPMY-1, but only on the morphology of prostate cancer PC-3 and DU145 cells. Compared with the control group, there were significant inhibitory effects on PC-3 and DU145 cell viability of prostate cancer at concentrations of 20 渭 mol / L and 100 渭 mol / L respectively. The nuclear breakdown rate of PC-3 cells was 27.0 卤2.0% and 63.0 卤3.0 卤3.0 卤3.0 卤3.0 卤3.0%, respectively, after treated with 20 渭 mol / L and 100 渭 mol / L kapotonin for 24 h, respectively, which was significantly higher than that in the blank control group (P = 3.0 卤1.0) and the apoptotic rates were 20.0 卤1.0 卤2.0% and 42.6 卤1.5, respectively, which were significantly higher than those in the blank control group (4.0 卤1.0). The expression levels of cleaved-PARP-1 cleaved-caspase-8 and cleaved-caspase 9 protein in PC-3 cells were significantly higher than those in the control group (P < 0.01), while the expression levels of XIAP survivin and Bcl-2 protein were significantly decreased by 0.001n.PC-3 in G _ 2 / M phase, and the expression levels of XIAPsurvivin and Bcl-2 protein were significantly decreased after treatment with 100 渭 mol 路L ~ (-1) of hirsucharin for 24 h, and the expression levels of cleaved-caspase-8 and cleaved-caspase _ 9 protein in PC-3 cells were significantly higher than those in the control group (P < 0.01). Moreover, the expression of cyclin B1 protein decreased significantly. Root dermatine inhibited the growth of xenografts in nude mice and up-regulated the expression of cleaved-PARP-1 and cleaved-caspase _ 3, and down-regulated the expression level of survivin protein (P = 0.001), but had no significant effect on the body mass of mice. Conclusion: by down-regulating the protein levels of cyclin B1-XIAPP, and up-regulating the expression of cleaved-PARP-1, cleaved-caspase-8 and cleaved-caspase 9, kaponin induces cell cycle arrest and apoptosis of PC-3 cells, and ultimately inhibits the growth of tumor cells.
【作者单位】：南京师范大学生命科学学院江苏省分子与医学生物技术重点实验室;
【基金】：国家自然科学基金资助项目(编号:81272850,81472415)~~
【分类号】：R737.25

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