S100A6蛋白对细胞中β-catenin水平的影响及可能机制

中国生物工程杂志

2011, Vol. 31

Issue (11): 18-23

研究报告

S100A6蛋白对细胞中β-catenin水平的影响及可能机制

黎玉叶, 李星星, 孙双双, 邹正渝, 张昀源, 段亮, 叶立伟, 武睿, 杨霞, 何通川, 周兰

重庆医科大学医学检验系临床检验诊断学教育部重点实验室重庆 400016

Effects and Possible Mechanism of Protein S100A6 on β-catenin

LI Yu-ye, LI Xing-xing, SUN Shuang-shuang, ZOU Zheng-yu, ZHANG Yun-yuan, DUAN Liang, YE Li-wei, WU Rui, YANG Xia, HE Tong-chuan, ZHOU Lan

Key Laboratory of Laboratory Medical Diagnostics, Department of Medical Laboratory, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

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摘要：

探讨S100A6蛋白对细胞中β-catenin水平的影响及可能机制。用表达S100A6及其siRNA的重组腺病毒AdS100A6和AdsiS100A6处理人骨肉瘤细胞系143B,Western blot分析处理前后细胞中β-catenin水平的变化;再以人胚肾细胞HEK293为研究对象,通过免疫共沉淀(co-immunoprecipitation,Co-IP)联合Western blot检测S100A6与Wnt经典信号通路主要成员β-catenin、糖原合酶激酶-3β(GSK-3β)、散乱蛋白(Dishevelled,DVL)和轴蛋白(Axin)之间的相互作用。结果:1)AdS100A6作用48h后,143B细胞中β-catenin水平较实验对照组(AdGFP组)增加了64%(P<0.01),而AdsiS100A6处理的细胞中β-catenin较AdGFP组减少了20.2%(P<0.05),AdGFP组与空白组之间的差异无统计学意义;2)在S100A6抗体的沉淀物中,检测到β-catenin、GSK-3β和DVL,而无Axin检出;3)β-catenin、GSK-3β和DVL的抗体沉淀物中,均有S100A6检出,而Axin抗体的沉淀物中无S100A6检出。结论:S100A6能够上调人骨肉瘤细胞系143B中的β-catenin水平;并与β-catenin、GSK-3β和DVL之间存在直接相互作用;S100A6与这三个分子之间的直接相互作用可能抑制了β-catenin的降解,使之在胞浆内聚集,水平升高。这可能是S100A6上调β-catenin的部分机制。

关键词： S100A6蛋白; Wnt经典信号通路; 蛋白质相互作用

Abstract:

The purpose is to analyze the effect of S100A6 on β-catenin and its possible molecular mechanisms. The recombinant adenovirus AdS100A6, AdsiS100A6 and AdGFP were used to infect human osteosarcoma cell line 143B respectively, then Western blot was used to detect cytoplasmic β-catenin levels. Adβ-catenin,pCMV-DVL-Myc,pCMV-Axin-Myc,pcDNA-GSK-3β-HA, pHAHA-S100A6 and pGST-S100A6 were used to infect or transfect or cotransfected HEK293 respectively, then co-immunoprecipitation and Western blot were used to investigate the interactions between S100A6 and β-catenin／GSK-3β／DVL／Axin. Results: 1) Compared with the control group(143B infected with AdGFP), the cytoplasmic β-catenin levels of S100A6 group(143B infected with AdS100A6) and siS100A6 group(143B infected with AdsiS100A6) increased by 64% (P<0.01) and decreased by 20.2%(P<0.05), respectively; there was no difference between control group and blank group; 2) β-catenin, GSK-3β and DVL were found in the precipitate of S100A6 antibody, but no Axin was found in it;3) S100A6 was found in precipitates of β-catenin antibody,GSK-3β antibody and DVL antibody,but not in precipitate of Axin antibody. The results suggest that S100A6 can upregulate the β-catenin level of human osteosarcoma cell line 143B and that there are direct interactions between S100A6 and β-catenin／GSK-3β／DVL. The interactions maybe inhibit the degradation of β-catenin and therefore increase its cytoplasmic level.

Key words: Protein S100A6 The canonical Wnt signaling pathway Protein-protein interaction

收稿日期: 2011-07-26 出版日期: 2011-11-25

ZTFLH:

Q51

基金资助:

国家自然科学基金资助项目(30772548)

通讯作者: 周兰 E-mail: zhoulan0111@foxmail.com

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引用本文:

黎玉叶, 李星星, 孙双双, 邹正渝, 张昀源, 段亮, 叶立伟, 武睿, 杨霞, 何通川, 周兰. S100A6蛋白对细胞中β-catenin水平的影响及可能机制[J]. 中国生物工程杂志, 2011, 31(11): 18-23.

LI Yu-ye, LI Xing-xing, SUN Shuang-shuang, ZOU Zheng-yu, ZHANG Yun-yuan, DUAN Liang, YE Li-wei, WU Rui, YANG Xia, HE Tong-chuan, ZHOU Lan. Effects and Possible Mechanism of Protein S100A6 on β-catenin. China Biotechnology, 2011, 31(11): 18-23.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I11/18

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