中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2008年
1期
12-15
,共4页
赖天霞%苗静琨%何焕玲%左国伟%李星星%王嫣%王胜%何通川%周兰
賴天霞%苗靜琨%何煥玲%左國偉%李星星%王嫣%王勝%何通川%週蘭
뢰천하%묘정곤%하환령%좌국위%리성성%왕언%왕성%하통천%주란
钙结合蛋白S100A6%Wnt/β-catenin信号途径%蛋白质相互作用
鈣結閤蛋白S100A6%Wnt/β-catenin信號途徑%蛋白質相互作用
개결합단백S100A6%Wnt/β-catenin신호도경%단백질상호작용
S100A6%Wnt/β-catenin signaling pathway%Protein-protein interaction
目的 研究钙结合蛋白S100A6对Wnt/β-catenin信号途径的影响及其机制.方法 异丙基β-D-硫代半乳糖苷(IPTG)诱导表达、谷胱甘肽-琼脂糖球珠分离纯化融合蛋白GST-hS100A6;Western blot和荧光素酶活性分析法检测S100A6对细胞内β-catenin水平和β-catenin/T细胞因子4(TCF4)活性的影响;GST-pulldown和Western blot技术研究S100A6与该信号途径主要成员β-catenin、糖原合酶激酶-3β(GSK-3β)、Dvl和Axin之间的相互作用.结果 S100A6可致MG63和HCT116细胞β-catenin水平升高,并可使HEK293细胞β-catenin/TCF4活性增强;S100A6与β-catenin、GSK-3β和Dvl之间存在着直接的相互作用,而与Axin之间无相互作用.结论 S100A6能够增强Wnt/β-catenin信号途径的活性,其机制可能涉及S100A6与该途径重要成员β-catenin、GSK-3β和Dvl之间存在着直接的相互作用有关.
目的 研究鈣結閤蛋白S100A6對Wnt/β-catenin信號途徑的影響及其機製.方法 異丙基β-D-硫代半乳糖苷(IPTG)誘導錶達、穀胱甘肽-瓊脂糖毬珠分離純化融閤蛋白GST-hS100A6;Western blot和熒光素酶活性分析法檢測S100A6對細胞內β-catenin水平和β-catenin/T細胞因子4(TCF4)活性的影響;GST-pulldown和Western blot技術研究S100A6與該信號途徑主要成員β-catenin、糖原閤酶激酶-3β(GSK-3β)、Dvl和Axin之間的相互作用.結果 S100A6可緻MG63和HCT116細胞β-catenin水平升高,併可使HEK293細胞β-catenin/TCF4活性增彊;S100A6與β-catenin、GSK-3β和Dvl之間存在著直接的相互作用,而與Axin之間無相互作用.結論 S100A6能夠增彊Wnt/β-catenin信號途徑的活性,其機製可能涉及S100A6與該途徑重要成員β-catenin、GSK-3β和Dvl之間存在著直接的相互作用有關.
목적 연구개결합단백S100A6대Wnt/β-catenin신호도경적영향급기궤제.방법 이병기β-D-류대반유당감(IPTG)유도표체、곡광감태-경지당구주분리순화융합단백GST-hS100A6;Western blot화형광소매활성분석법검측S100A6대세포내β-catenin수평화β-catenin/T세포인자4(TCF4)활성적영향;GST-pulldown화Western blot기술연구S100A6여해신호도경주요성원β-catenin、당원합매격매-3β(GSK-3β)、Dvl화Axin지간적상호작용.결과 S100A6가치MG63화HCT116세포β-catenin수평승고,병가사HEK293세포β-catenin/TCF4활성증강;S100A6여β-catenin、GSK-3β화Dvl지간존재착직접적상호작용,이여Axin지간무상호작용.결론 S100A6능구증강Wnt/β-catenin신호도경적활성,기궤제가능섭급S100A6여해도경중요성원β-catenin、GSK-3β화Dvl지간존재착직접적상호작용유관.
Objective To analyze the effects of S100A6 on Wnt/β-catenin signaling pathway and its molecular mechanism. Methods The expression of GST-hS10OA6 was induced with IPTG in Escherichia coli BL21, and the fusion protein was purified with glutathione-sepharose 4B beads. β-catenin level of human colon cancer cell line MG63 and human osteosarcoma cell line HCTl16 cells infected with AdS10OA6 was measured by Western blot. Luciferase activity assay was applied to analyze the effect of S100A6 on the β-catenin/TCF4 activity. The interactions between S100A6 and β-catenin/GSK-3β/Dvl/Axin were detected by GST-pulldown/Western blot. Results The β-catenin level in AdS100A6-infected MG63 and HCT116 cells was significantly increased in comparison with that in the AdGFP control group (P<0.01). The luciferase activity in human embryonic renal cell line 293 cells transfected with pTOP-Luc and followed by GST-hS100A6 treatment was increased by 20. 2-fold in comparison with that in the GST control group (P<0.01). The interaction between GST-hS100A6 and Axin was not found. Conclusion S100A6 up- regulates the Wnt/β-catenin signaling pathway, and this may be attributed to the interaction between S100A6 and β-catenin/GSK-3β/Dvl.
