【摘要】：目的： 了解曲古抑菌素A（trichostatinA，TSA）對人膀胱癌BIU-87細(xì)胞PTEN(phosphatase and tensin homolog deleted on chromosome ten)基因和Runx3（humanrunt2related transcription fac2tor3）表達(dá)的影響，并探討曲古抑菌素A作為治療膀胱癌新化療藥的可行性及可能的相關(guān)作用機(jī)制。 方法： 取對數(shù)生長期的BIU-87細(xì)胞，按照一定的細(xì)胞培養(yǎng)方法制成單細(xì)胞懸液，用含有不同濃度藥物的培養(yǎng)基作為藥物組，培養(yǎng)一定時間后用噻唑藍(lán)（MTT）法檢測膀胱癌BIU-87細(xì)胞的存活率，觀察TSA對BIU-87細(xì)胞存活率的影響；用半定量RT-PCR法探討抑癌基因PTEN mRNA、RUNX3mRNA水平改變情況；用Western-blotting法觀察抑癌基因PTEN蛋白、RUNX3蛋白的表達(dá)。 結(jié)果： 1.以TSA濃度0nmol/L作為正常組來參照，以濃度為150nmol/L、300n mol/L、600nmol/L、1200nmol/L的TSA作為處理組，作用于BIU-87細(xì)胞分別處理24、48和72h后，每時間組內(nèi)部細(xì)胞存活率明顯降低，，細(xì)胞活性具有劑量相關(guān)性下降。當(dāng)TSA濃度為600nmol/L和1200nmol/L時，分別處理BIU-87細(xì)胞24、48和72h后，細(xì)胞活性呈時間相關(guān)性降低（P0.05，P0.01）。 2.半定量RT-PCR結(jié)果，與對照組比較TSA在300nmol/L～1200nmol/L作用72h后使膀胱癌BIU-87細(xì)胞PTEN mRNA、RUNX3mRNA表達(dá)升高，且二者都表現(xiàn)出劑量相關(guān)性上升（P0.05，P0.01）。 3. Western-blotting結(jié)果，與對照組相比TSA在300nmol/L～1200nmol/L處理72h后使膀胱癌BIU-87細(xì)胞PTEN蛋白和RUNX3蛋白表達(dá)上調(diào)，且二者分別具有劑量相關(guān)性上升（P0.05，P0.01）。 結(jié)論： 1. TSA以150nmol/L～1200nmol/L濃度處理時，對BIU 87細(xì)胞增殖抑制表現(xiàn)為劑量依賴性；同時TSA濃度為150nmol/L和1200nmol/L之間時，其對BIU 87細(xì)胞增殖抑制還存在時間依賴性。 2. TSA可以誘使膀胱癌BIU 87細(xì)胞凋亡且在濃度150nmol/L和1200nmol/L之間凋亡率具有劑量和時間相關(guān)性。 3. TSA促進(jìn)膀胱癌BIU 87細(xì)胞凋亡的作用機(jī)制可能與上調(diào)抑癌基PTENmRNA、RUNX3mRNA的轉(zhuǎn)錄水平以及兩者的蛋白的表達(dá)量來實(shí)現(xiàn)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.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.14

【參考文獻(xiàn)】

中國期刊全文數(shù)據(jù)庫 前7條

1 張海元;云鵬;;肝癌細(xì)胞系中Runx3基因表達(dá)及啟動子區(qū)異常甲基化分析[J];長江大學(xué)學(xué)報(bào)(自科版)醫(yī)學(xué)卷;2009年01期

2 高鋒利;王海泉;劉瑞;李昭宇;;RUNX3在人類原發(fā)性肝細(xì)胞癌中的表達(dá)[J];寧夏醫(yī)學(xué)雜志;2010年06期

3 陳秀娟;鄧濤;羅和生;;RUNX3、Bcl-2和bax蛋白在大腸癌中的表達(dá)及其意義[J];胃腸病學(xué)和肝病學(xué)雜志;2009年03期

4 張春艷;劉志方;徐霞;曾季平;于晗;賈繼輝;;腫瘤抑制因子RUNX3對人胃癌細(xì)胞中凋亡相關(guān)基因表達(dá)的影響[J];中國病理生理雜志;2009年07期

5 溫機(jī)靈;周祥福;溫機(jī)智;周琦;王林;;抑癌基因RUNX3在膀胱癌T24細(xì)胞株中的表達(dá)以及對凋亡的影響[J];中華腔鏡泌尿外科雜志(電子版);2008年04期

6 李功成,張旭,李龍承,葉章群;曲古抑菌素A對膀胱癌細(xì)胞的抑制作用及其機(jī)制[J];中國現(xiàn)代醫(yī)學(xué)雜志;2005年07期

7 楊娜;李智;段建敏;徐耀庭;;膀胱癌組織RUNX3基因的甲基化改變[J];腫瘤;2008年08期

本文編號：2424511