Ye Xiong, Fu Qiang, Xiao Hui
School of Clinical Medicine, Shanghai University of Medicine & Health Sciences, Shanghai, China.
Department of Respiratory and Critical Care Medicine, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai, China.
Front Genet. 2022 Apr 5;13:813268. doi: 10.3389/fgene.2022.813268. eCollection 2022.
The overexpression of human antigen R (HuR) has been proven in various types of cancer and is associated with the poor survival lung cancer patients. HuR overexpression stabilizes the mRNA of tumor-promoting genes by binding with 3'-UTR AU-rich elements. However, the role of HuR in the proliferation of lung cancer is unclear. HuR expression was assessed using immunohistochemistry of tumor tissue samples from ten patients with lung cancer and ten patients with benign lung disease. Gene, protein, mRNA, and lncRNA changes in A549 HuR knockdown (KD) cells were assessed by single-cell RNA sequencing analysis. Furthermore, cell proliferation, migration, and invasion were determined by Cell Counting Kit-8 (CCK-8) assays and Transwell assays with or without Matrigel. The cell cycle was assessed by propidium iodide staining. The protein level, mRNA level and half-life of PLK1 were detected by western blotting and RT-qPCR. In clinical patients, the expression of HuR was significantly higher in lung cancer patients than in patients with benign lung disease. RNA sequencing analysis of A549 HuR knockdown cells revealed that the main function of HuR was related to ribonucleoprotein complex biogenesis. HuR was found to regulate signaling pathways mainly related to the spliceosome, RNA transport and the cell cycle. HuR KD suppressed the proliferation, migration and invasion of A549 cells, indicating its promotive role in these processes. These results demonstrate that HuR plays an important role in the progression of lung cancer.
人类抗原R（HuR）的过表达已在多种类型的癌症中得到证实，并且与肺癌患者的不良生存状况相关。HuR过表达通过与3'-UTR富含AU的元件结合来稳定促癌基因的mRNA。然而，HuR在肺癌增殖中的作用尚不清楚。使用免疫组织化学方法对10例肺癌患者和10例良性肺病患者的肿瘤组织样本进行HuR表达评估。通过单细胞RNA测序分析评估A549 HuR敲低（KD）细胞中的基因、蛋白质、mRNA和lncRNA变化。此外，通过细胞计数试剂盒-8（CCK-8）检测以及有或没有基质胶的Transwell检测来确定细胞增殖、迁移和侵袭。通过碘化丙啶染色评估细胞周期。通过蛋白质印迹和RT-qPCR检测PLK1的蛋白质水平、mRNA水平和半衰期。在临床患者中，肺癌患者的HuR表达明显高于良性肺病患者。对A549 HuR敲低细胞的RNA测序分析表明，HuR的主要功能与核糖核蛋白复合体生物合成有关。发现HuR主要调节与剪接体、RNA转运和细胞周期相关的信号通路。HuR KD抑制了A549细胞的增殖、迁移和侵袭，表明其在这些过程中起促进作用。这些结果表明，HuR在肺癌进展中起重要作用。
