曲古抑菌素A对膀胱癌BIU-87细胞PTEN和RUNX3基因表达的影响
发布时间：2019-02-16 13:48
【摘要】：目的： 了解曲古抑菌素A（trichostatinA，TSA）对人膀胱癌BIU-87细胞PTEN(phosphatase and tensin homolog deleted on chromosome ten)基因和Runx3（humanrunt2related transcription fac2tor3）表达的影响，并探讨曲古抑菌素A作为治疗膀胱癌新化疗药的可行性及可能的相关作用机制。 方法： 取对数生长期的BIU-87细胞，按照一定的细胞培养方法制成单细胞悬液，用含有不同浓度药物的培养基作为药物组，培养一定时间后用噻唑蓝（MTT）法检测膀胱癌BIU-87细胞的存活率，观察TSA对BIU-87细胞存活率的影响；用半定量RT-PCR法探讨抑癌基因PTEN mRNA、RUNX3mRNA水平改变情况；用Western-blotting法观察抑癌基因PTEN蛋白、RUNX3蛋白的表达。 结果： 1.以TSA浓度0nmol/L作为正常组来参照，以浓度为150nmol/L、300n mol/L、600nmol/L、1200nmol/L的TSA作为处理组，作用于BIU-87细胞分别处理24、48和72h后，每时间组内部细胞存活率明显降低，，细胞活性具有剂量相关性下降。当TSA浓度为600nmol/L和1200nmol/L时，分别处理BIU-87细胞24、48和72h后，细胞活性呈时间相关性降低（P0.05，P0.01）。 2.半定量RT-PCR结果，与对照组比较TSA在300nmol/L～1200nmol/L作用72h后使膀胱癌BIU-87细胞PTEN mRNA、RUNX3mRNA表达升高，且二者都表现出剂量相关性上升（P0.05，P0.01）。 3. Western-blotting结果，与对照组相比TSA在300nmol/L～1200nmol/L处理72h后使膀胱癌BIU-87细胞PTEN蛋白和RUNX3蛋白表达上调，且二者分别具有剂量相关性上升（P0.05，P0.01）。 结论： 1. TSA以150nmol/L～1200nmol/L浓度处理时，对BIU 87细胞增殖抑制表现为剂量依赖性；同时TSA浓度为150nmol/L和1200nmol/L之间时，其对BIU 87细胞增殖抑制还存在时间依赖性。 2. TSA可以诱使膀胱癌BIU 87细胞凋亡且在浓度150nmol/L和1200nmol/L之间凋亡率具有剂量和时间相关性。 3. TSA促进膀胱癌BIU 87细胞凋亡的作用机制可能与上调抑癌基PTENmRNA、RUNX3mRNA的转录水平以及两者的蛋白的表达量来实现的。
[Abstract]:Objective: to investigate the effect of trigostatin A (trichostatinA,TSA) on the expression of PTEN (phosphatase and tensin homolog deleted on chromosome ten) gene and Runx3 (humanrunt2related transcription fac2tor3) in human bladder cancer BIU-87 cells. To explore the feasibility and mechanism of trigostatin A as a new chemotherapeutic agent for bladder cancer. Methods: BIU-87 cells in logarithmic growth period were collected and prepared into single cell suspension according to a certain cell culture method. The culture medium containing different concentrations of drugs was used as the drug group. The survival rate of bladder cancer BIU-87 cells was detected by thiazolyl (MTT) assay for a certain time, and the effect of TSA on the survival rate of BIU-87 cells was observed. The changes of tumor suppressor gene PTEN mRNA,RUNX3mRNA and the expression of PTEN protein and RUNX3 protein were studied by semi-quantitative RT-PCR and Western-blotting method respectively. Results: 1. Compared with the normal group with TSA concentration 0nmol/L and the TSA treated with 150nmol / L ~ (300n) mol/L,600nmol/L,1200nmol/L as the treatment group, the survival rate of BIU-87 cells in each time group was significantly decreased after treated with BIU-87 cells for 24 ~ 48 h and 72 h, respectively. Cell activity decreased in a dose-dependent manner. When the concentration of TSA was 600nmol/L and 1200nmol/L, the activity of BIU-87 cells decreased in a time-dependent manner (P0.05, P0.01) after 24 h and 72 h treatment, respectively. 2. Compared with the control group, TSA increased the expression of PTEN mRNA,RUNX3mRNA in BIU-87 cells of bladder cancer after 72 h of 300nmol/L~1200nmol/L treatment, and both of them showed a dose-dependent increase (P0.05, P0.01). 3. Western-blotting results showed that TSA increased the expression of PTEN protein and RUNX3 protein in BIU-87 cells of bladder cancer at 72 h after 300nmol/L~1200nmol/L treatment, and the expression of PTEN protein and RUNX3 protein increased in a dose-dependent manner (P0.05, P0.01). Conclusion: 1. When TSA was treated with 150nmol/L~1200nmol/L, the inhibition of proliferation of BIU Thun87 cells was dose-dependent, while that of TSA between 150nmol/L and 1200nmol/L was time-dependent. 2. TSA could induce apoptosis of bladder cancer BIU Thun87 cells, and the apoptotic rate between 150nmol/L and 1200nmol/L was dose-dependent and time-dependent. 3. The mechanism of TSA in promoting apoptosis of bladder cancer cell line BIU O87 may be related to the up-regulation of the transcription level of tumor suppressor PTENmRNA,RUNX3mRNA and the expression of both proteins.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14
本文编号：2424511
[Abstract]:Objective: to investigate the effect of trigostatin A (trichostatinA,TSA) on the expression of PTEN (phosphatase and tensin homolog deleted on chromosome ten) gene and Runx3 (humanrunt2related transcription fac2tor3) in human bladder cancer BIU-87 cells. To explore the feasibility and mechanism of trigostatin A as a new chemotherapeutic agent for bladder cancer. Methods: BIU-87 cells in logarithmic growth period were collected and prepared into single cell suspension according to a certain cell culture method. The culture medium containing different concentrations of drugs was used as the drug group. The survival rate of bladder cancer BIU-87 cells was detected by thiazolyl (MTT) assay for a certain time, and the effect of TSA on the survival rate of BIU-87 cells was observed. The changes of tumor suppressor gene PTEN mRNA,RUNX3mRNA and the expression of PTEN protein and RUNX3 protein were studied by semi-quantitative RT-PCR and Western-blotting method respectively. Results: 1. Compared with the normal group with TSA concentration 0nmol/L and the TSA treated with 150nmol / L ~ (300n) mol/L,600nmol/L,1200nmol/L as the treatment group, the survival rate of BIU-87 cells in each time group was significantly decreased after treated with BIU-87 cells for 24 ~ 48 h and 72 h, respectively. Cell activity decreased in a dose-dependent manner. When the concentration of TSA was 600nmol/L and 1200nmol/L, the activity of BIU-87 cells decreased in a time-dependent manner (P0.05, P0.01) after 24 h and 72 h treatment, respectively. 2. Compared with the control group, TSA increased the expression of PTEN mRNA,RUNX3mRNA in BIU-87 cells of bladder cancer after 72 h of 300nmol/L~1200nmol/L treatment, and both of them showed a dose-dependent increase (P0.05, P0.01). 3. Western-blotting results showed that TSA increased the expression of PTEN protein and RUNX3 protein in BIU-87 cells of bladder cancer at 72 h after 300nmol/L~1200nmol/L treatment, and the expression of PTEN protein and RUNX3 protein increased in a dose-dependent manner (P0.05, P0.01). Conclusion: 1. When TSA was treated with 150nmol/L~1200nmol/L, the inhibition of proliferation of BIU Thun87 cells was dose-dependent, while that of TSA between 150nmol/L and 1200nmol/L was time-dependent. 2. TSA could induce apoptosis of bladder cancer BIU Thun87 cells, and the apoptotic rate between 150nmol/L and 1200nmol/L was dose-dependent and time-dependent. 3. The mechanism of TSA in promoting apoptosis of bladder cancer cell line BIU O87 may be related to the up-regulation of the transcription level of tumor suppressor PTENmRNA,RUNX3mRNA and the expression of both proteins.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14
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本文编号：2424511
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