中国肿瘤临床
中國腫瘤臨床
중국종류림상
CHINESE JOURNAL OF CLINICAL ONCOLOGY
2013年
1期
16-20
,共5页
赵恩宏%许倩%肖丽君%刘镭%赵爽%高亚贤%郑鑫
趙恩宏%許倩%肖麗君%劉鐳%趙爽%高亞賢%鄭鑫
조은굉%허천%초려군%류뢰%조상%고아현%정흠
17-AAG%细胞增殖%STAT3%VEGF
17-AAG%細胞增殖%STAT3%VEGF
17-AAG%세포증식%STAT3%VEGF
17-AAG%cell proliferation%VEGF%STAT3
目的:观察17-烯丙胺-17-脱甲氧格尔德霉素(17-allylamino-17-desmethoxy-geldanamycin,17-AAG)对人胃癌MKN-45细胞信号转导和转录活化蛋白3(signal transducer and activator of transcriptions 3,STAT3)、血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA 及蛋白表达水平的影响,并探寻 STAT3通路所发挥的作用.方法:体外培养人胃癌MKN-45细胞,分别给予不同剂量及不同作用时间17-AAG进行作用,四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞增殖能力;RT-PCR法和Western Blotting法检测各组细胞STAT3和VEGF mRNA和蛋白的表达水平.结果:0.165~10 mg/L的17-AAG作用24、48 h后对MKN-45细胞有显著的抑制作用,且有明显的时间、剂量依赖性;1.0、2.0、3.0、5.0 mg/L的17-AAG作用48 h后,各组细胞STAT3和VEGF mRNA和蛋白表达均下调,且具有浓度依赖性;3.0 mg/L 17-AAG作用12、24、48h后,各组细胞STAT3和VEGF mRNA和蛋白表达均下调,且具有时间依赖性.结论:17-AAG对人胃癌MKN-45细胞的增殖具有明显的抑制作用,并能抑制STAT3和VEGF mRNA和蛋白的表达,17-AAG可能通过对STAT3通路的负性调控作用而抑制VEGF表达.
目的:觀察17-烯丙胺-17-脫甲氧格爾德黴素(17-allylamino-17-desmethoxy-geldanamycin,17-AAG)對人胃癌MKN-45細胞信號轉導和轉錄活化蛋白3(signal transducer and activator of transcriptions 3,STAT3)、血管內皮生長因子(vascular endothelial growth factor,VEGF)mRNA 及蛋白錶達水平的影響,併探尋 STAT3通路所髮揮的作用.方法:體外培養人胃癌MKN-45細胞,分彆給予不同劑量及不同作用時間17-AAG進行作用,四甲基偶氮唑鹽微量酶反應比色法(MTT法)檢測細胞增殖能力;RT-PCR法和Western Blotting法檢測各組細胞STAT3和VEGF mRNA和蛋白的錶達水平.結果:0.165~10 mg/L的17-AAG作用24、48 h後對MKN-45細胞有顯著的抑製作用,且有明顯的時間、劑量依賴性;1.0、2.0、3.0、5.0 mg/L的17-AAG作用48 h後,各組細胞STAT3和VEGF mRNA和蛋白錶達均下調,且具有濃度依賴性;3.0 mg/L 17-AAG作用12、24、48h後,各組細胞STAT3和VEGF mRNA和蛋白錶達均下調,且具有時間依賴性.結論:17-AAG對人胃癌MKN-45細胞的增殖具有明顯的抑製作用,併能抑製STAT3和VEGF mRNA和蛋白的錶達,17-AAG可能通過對STAT3通路的負性調控作用而抑製VEGF錶達.
목적:관찰17-희병알-17-탈갑양격이덕매소(17-allylamino-17-desmethoxy-geldanamycin,17-AAG)대인위암MKN-45세포신호전도화전록활화단백3(signal transducer and activator of transcriptions 3,STAT3)、혈관내피생장인자(vascular endothelial growth factor,VEGF)mRNA 급단백표체수평적영향,병탐심 STAT3통로소발휘적작용.방법:체외배양인위암MKN-45세포,분별급여불동제량급불동작용시간17-AAG진행작용,사갑기우담서염미량매반응비색법(MTT법)검측세포증식능력;RT-PCR법화Western Blotting법검측각조세포STAT3화VEGF mRNA화단백적표체수평.결과:0.165~10 mg/L적17-AAG작용24、48 h후대MKN-45세포유현저적억제작용,차유명현적시간、제량의뢰성;1.0、2.0、3.0、5.0 mg/L적17-AAG작용48 h후,각조세포STAT3화VEGF mRNA화단백표체균하조,차구유농도의뢰성;3.0 mg/L 17-AAG작용12、24、48h후,각조세포STAT3화VEGF mRNA화단백표체균하조,차구유시간의뢰성.결론:17-AAG대인위암MKN-45세포적증식구유명현적억제작용,병능억제STAT3화VEGF mRNA화단백적표체,17-AAG가능통과대STAT3통로적부성조공작용이억제VEGF표체.
Objective: This study aims to investigate the effects of 17-allylamino -17-desmethoxy-geldanamycin (17-AAG) on the gene and protein expressions of VEGF and STAT3 pathways in MKN-45 human gastric cancer cells, and explore the potential mechanisms of the STAT3 signaling pathway that mediate these effects. Methods: MKN-45 cells were seeded in Dulbecco's Modified Eagle Medium. The thiazolyl blue method, i. e., methyl thiazolyl tetrazolium assay, was performed to evaluate the inhibitory effects of 17-AAG on the proliferation of MKN-45 cells at different times, with different doses. The gene and protein expression of VEGF and STAT3 were determined by reverse transcription polymerase chain reaction (RT–PCR) and Western blot analysis. Results: MKN-45 cells were treated with 17-AAG at 0.165 mg/L to 10 mg/L for 24 and 48 h, respectively. 17-AAG significantly inhibited MKN-45 cell proliferation in a time- and dose-dependent manner. The results of the RT–PCR and Western blot assays showed that the gene and protein expressions of VEGF and STAT3 in MKN-45 cells induced by 17-AAG were significantly down-regulated in a dose-dependent manner when the cells were treated with 17-AAG at 1.0, 2.0, 3.0, and 5.0 mg/L for 48 h. The gene and protein expressions of VEGF and STAT3 in the MKN-45 cells were significantly down-regulated in a time-dependent manner when the cells were treated with 17-AAG at 3.0 mg/L for 12, 24, and 48 h. Conclusion: The drug 17-AAG can inhibit cell proliferation and down-regulate the gene and protein expressions of VEGF and STAT3 in MKN-45 human gastric cancer cells in vitro; the down-regulation of VEGF expression may be associated with the STAT3 signaling pathway.