上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (1): 28-.doi: 10.3969/j.issn.1674-8115.2019.01.006

• 论著·基础研究 • 上一篇    下一篇

PDGFRα下游新底物 CCT2对肿瘤细胞生长的影响及其机制

屈国君 1，陆元凤 2，李宇 1   

  1. 1.上海交通大学基础医学院免疫学与微生物学系，上海 200025；2.上海交通大学医学院附属瑞金医院胃肠外科，上海 200025
  • 出版日期:2019-01-28 发布日期:2019-02-27
  • 通讯作者: 李宇，电子信箱：yulihunan@126.com。
  • 作者简介:屈国君（1989—），女，助理实验师，硕士；电子信箱：xiaoguojun@126.com。
  • 基金资助:
    上海市卫生和计划生育委员会青年项目（ 20144Y0071）

Mechanism of CCT2, a new downstream substrate of PDGFRα, on proliferation of tumor cells

QU Guo-jun1, LU Yuan-feng2, LI Yu1   

  1. 1. Department of Immunology and Microbiology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 2. Department of Gastrointestinal Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2019-01-28 Published:2019-02-27
  • Supported by:
    Youth Program of Shanghai Municipal Commission of Health and Family Planning, 20144Y0071

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摘要： 目的 ·研究血小板源性生长因子受体 α（platelet derived growth factor receptor α，PDGFRα）的下游新靶点分子伴侣复合体 β亚基（ chaperonin containing TCP1 subunit 2，CCT2）对肿瘤细胞生长的影响及其机制。方法 ·选取非小细胞肺癌细胞系 H1703作为细胞模型。采用 Western blotting方法检测 H1703细胞在 PDGFRα抑制剂 Gleevec处理以及 PDGF刺激条件下 CCT2的磷酸化；将 H1703细胞分为对照组（ siCon组）、 siPDGFRα组和 siCCT2组，分别转染 siRNA，待 48 h后对细胞进行计数；将 H1703细胞分为对照组（ siCon组）、 siPDGFRα组、siAKT组和 siCCT2组，采用 Western blotting方法检测 4组细胞中 PDGFRα和 PARP蛋白水平；采用细胞组分分离方法检测 CCT2在 H1703细胞中的定位，通过免疫共沉淀方法检测 CCT2 与 PDGFRα的相互作用。结果 · Gleevec抑制 CCT2 的磷酸化， PDGF诱导 CCT2磷酸化；与对照组比较， siCCT2转染的细胞数量降低了约 30%（P0.006）；与对照组比较， siCCT2转染的细胞中 PDGFRα蛋白表达量减少， PARP切割增加； CCT2在细胞膜类组分和细胞质中都有分布，并可以与 PDGFRα相互作用。结论 · PDGFRα下游新靶点 CCT2通过与 PDGFRα相互作用，维持 PDGFRα蛋白稳定性，从而促进 H1703细胞生长。

关键词: 血小板源性生长因子受体&, alpha, 非小细胞肺癌, 分子伴侣复合体&, beta, 亚基, 细胞生长, 蛋白稳定性

Abstract:

Objective · To study the molecular mechanism of chaperonin containing TCP1 subunit 2(CCT2), a new downstream substrate of platelet derived growth factor receptor α(PDGFRα), in tumorigenesis. Methods · Non-small cell lung cancer cell line H1703 was used. Western blotting was used to measure the phosphorylation of CCT2 upon PDGFRα inhibitor Gleevec treatment and PDGF stimulation. H1703 cells were divided into siCon group, siPDGFRα group and siCCT2 group; 48 h later, cell number counting was used to test the effect of CCT2 on cell growth after siRNA transfection. H1703 cells were divided into siCon group, siPDGFRα group, siAKT group and siCCT2 group; Western blotting was used to measure the protein level of PDGFRα and PARP. Cell fractionation was used to detect the cellular localization of CCT2 and co-immunoprecipitation was used to test the interaction between CCT2 and PDGFRα. Results · CCT2 phosphorylation was inhibitedGleevec and inducedPDGF. Compared to the control group, the number of cells transfectedsiCCT2 reduced30% (P0.006). The protein level of PDGFRα was also decreased in siCCT2 transfected cells, whereas the cleavage of PARP was increased. CCT2 was localized in both cytoplasmic and membrane fractions and interacted with PDGFRα directly. Conclusion · CCT2 is a new downstream substrate of PDGFRα. CCT2 can promote tumor cells growthinteracting and stabilizing PDGFRα.

Key words: platelet derived growth factor receptor &, alpha, non-small cell lung cancer, chaperonin containing TCP1 subunit 2, cell growth, protein stability

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