發(fā)布時間：2018-06-16 12:33

  本文選題：前列腺腫瘤 + 根皮素　； 參考：《腫瘤》2017年06期

【摘要】：目的 :探討根皮素對前列腺癌PC-3細胞凋亡和周期的影響,及對其裸鼠移植瘤生長的影響。方法 :不同濃度的根皮素(0、20、50和100μmol/L)分別作用于人正常前列腺基質(zhì)細胞WPMY-1和前列腺癌PC-3和DU145細胞24 h后,倒置光學顯微鏡下觀察細胞形態(tài)的變化,并采用CCK-8法檢測各組細胞活性的變化。用不同濃度的根皮素(0、20、50和100μmol/L)處理PC-3細胞24 h后,采用DAPI染色法觀察細胞核破裂的情況,FCM法檢測根皮素處理后前列腺癌PC-3細胞凋亡率和細胞周期分布的變化;同時,采用蛋白質(zhì)印跡法檢測細胞凋亡相關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連鎖凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)、survivin、Bcl-2、Bax以及細胞周期蛋白cyclin B1表達水平的變化。建立人前列腺癌PC-3細胞裸鼠皮下移植瘤模型,觀察根皮素腹腔注射治療后小鼠移植瘤生長體積以及體質(zhì)量的變化,并計算抑瘤率。采用蛋白質(zhì)印跡法檢測小鼠腫瘤組織中凋亡相關蛋白的表達水平。結果:根皮素濃度為50和100μmol/L時,對人正常前列腺基質(zhì)細胞WPMY-1的形態(tài)沒有影響,僅對前列腺癌PC-3和DU145細胞的形態(tài)有影響。與空白對照組相比,根皮素濃度為20、50和100μmol/L時對前列腺癌PC-3和DU145細胞活力均有明顯的抑制作用(P值均0.01)。采用濃度為20、50和100μmol/L的根皮素處理PC-3細胞24 h后,PC-3細胞的核破裂率分別為(27.0±2.0)%、(47.0±1.5)%和(63.0±3.0)%,明顯高于空白對照組的(3.0±1.0)%(P值均0.001);凋亡率分別為(20.0±1.0)%、(32.0±2.0)%和(42.6±1.5)%,明顯高于空白對照組的(4.0±1.0)%(P值均0.001)。根皮素(100μmol/L)處理PC-3細胞24 h后,細胞中cleaved-PARP-1、cleaved-caspase 3、cleaved-caspase 8和cleaved-caspase 9蛋白的表達水平均較空白對照組明顯升高(P值均0.01),而XIAP、survivin和Bcl-2蛋白的表達水平均明顯降低(P值均0.001)。PC-3細胞被阻滯在G2/M期,并且cyclin B1蛋白的表達水平明顯降低(P0.001)。根皮素可以抑制裸鼠移植瘤的生長(P0.05),并上調(diào)腫瘤組織中cleaved-PARP-1和cleaved-caspase 3蛋白的表達水平,下調(diào)survivin蛋白的表達水平(P值均0.001),但是對小鼠體質(zhì)量無明顯影響(P0.05)。結論 :根皮素通過下調(diào)cyclin B1、XIAP、survivin和Bcl-2蛋白水平,以及上調(diào)cleaved-PARP-1、cleaved-caspase 3、cleaved-caspase 8和cleaved-caspase 9蛋白的表達水平,誘導PC-3細胞周期阻滯和細胞凋亡,最終抑制腫瘤細胞生長。
[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 鹵2.0% and 63.0 鹵3.0 鹵3.0 鹵3.0 鹵3.0 鹵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 鹵1.0) and the apoptotic rates were 20.0 鹵1.0 鹵2.0% and 42.6 鹵1.5, respectively, which were significantly higher than those in the blank control group (4.0 鹵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.
【作者單位】： 南京師范大學生命科學學院江蘇省分子與醫(yī)學生物技術重點實驗室;
【基金】：國家自然科學基金資助項目(編號:81272850,81472415)~~
【分類號】：R737.25

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