School of Life Sciences, Shanxi University, Taiyuan, Shanxi, China.
Central Laboratory, Linyi People's Hospital, Linyi, Shandong, China.
J Cell Physiol. 2019 Jul;234(7):11511-11523. doi: 10.1002/jcp.27813. Epub 2018 Nov 27.
We have recently reported that type 2 diabetes promotes centrosome amplification via enhancing the expression, biding, and centrosome translocation of rho-associated coiled-coil containing protein kinase 1 (ROCK1)/14-3-3σ complex in HCT116 cells. In the functional proteomic study, we further investigated the molecular pathways underlying the centrosome amplification using HCT116 cells. We found that treatment of HCT116 cells with high glucose, insulin, and palmitic acid triggered the centrosome amplification and increased the expressions of proliferating cell nuclear antigen (PCNA), nucleophosmin (NPM), and 14-3-3σ. Individual knockdown of PCNA, NPM, or 14-3-3σ inhibited the centrosome amplification. Knockdown of PCNA inhibited the treatment-increased expression of ROCK1, whereas knockdown of ROCK1 did not affect the PCNA expression. High glucose, insulin, and palmitic acid also increased the expressions of c-Jun N-terminal kinase-1 (JNK1) and signal transducer and activator of transcription 3 (Stat3), individual knockdown of which upregulated the treatment-increased expression of 14-3-3σ and promoted the centrosome amplification. In contrast, overexpression of JNK1 inhibited the centrosome amplification. Knockdown of Stat3 enhanced the centrosome translocation of 14-3-3σ. Moreover, we showed that knockdown of JNK1 inhibited the treatment-increased expression of Stat3. Knockdown of PCNA, JNK1, or Stat3 did not have an effect on NPM and vice versa. In conclusion, our results suggest that PCNA and JNK1-Stat3 pathways respectively promotes and feedback inhibits the centrosome amplification by targeting at the ROCK1/14-3-3σ complex, and NPM serves as an independent signal for the centrosome amplification.
我们最近报道，2 型糖尿病通过增强 Rho 相关卷曲螺旋蛋白激酶 1（ROCK1）/14-3-3σ复合物在 HCT116 细胞中的表达、结合和向中心体易位，促进中心体扩增。在功能蛋白质组学研究中，我们使用 HCT116 细胞进一步研究了中心体扩增的分子途径。我们发现，用高葡萄糖、胰岛素和棕榈酸处理 HCT116 细胞会触发中心体扩增，并增加增殖细胞核抗原（PCNA）、核仁磷酸蛋白（NPM）和 14-3-3σ 的表达。单独敲低 PCNA、NPM 或 14-3-3σ 可抑制中心体扩增。敲低 PCNA 抑制了治疗增加的 ROCK1 表达，而敲低 ROCK1 并不影响 PCNA 的表达。高葡萄糖、胰岛素和棕榈酸也增加了 c-Jun N-末端激酶-1（JNK1）和信号转导和转录激活因子 3（Stat3）的表达，单独敲低 JNK1 或 Stat3 均可上调治疗增加的 14-3-3σ 的表达并促进中心体扩增。相反，JNK1 的过表达抑制了中心体扩增。敲低 Stat3 增强了 14-3-3σ 的中心体易位。此外，我们还表明，敲低 JNK1 抑制了 Stat3 的治疗增加表达。敲低 PCNA、JNK1 或 Stat3 对 NPM 没有影响，反之亦然。总之，我们的结果表明，PCNA 和 JNK1-Stat3 途径分别通过靶向 ROCK1/14-3-3σ 复合物促进和反馈抑制中心体扩增，而 NPM 则作为中心体扩增的独立信号。
