Chinese Medical Sciences Journal ›› 2018, Vol. 33 ›› Issue (3): 143-151.doi: 10.24920/11815

• Original Article • Previous Articles     Next Articles

RNA-binding Protein UNR Promotes Glioma Cell Migration and Regulates the Expression of Ribosomal Protein L9

Tian Ningyu, Qi Yingjiao, Hu Yan, Yin Bin, Yuan Jiangang, Qiang Boqin, Peng Xiaozhong(), Han Wei()   

  1. State Key Laboratory of Medical Molecular Biology, Department of Molecular Biology and Biochemistry, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences & School of Basic Medicine Peking Union Medical College, Beijing 100005, China
  • Published:2018-09-30 Online:2018-06-01
  • Contact: Peng Xiaozhong,Han Wei E-mail:hanwei2012@ibms.pumc.edu.cn;pengxiaozhong@pumc.edu.cn
The authors transfected UNR siRNAs in glioma cells and observed cell proliferation and migration, and scratch wound-healing assay revealing knockdown of UNR dramatically inhibited the cell migration.

Objective To investigate the role of RNA binding protein—upstream-of-N-Ras (UNR) in the development of glioma and its molecular mechanism.Methods First, bioinformatics analysis of CGGA database was performed to detect UNR expression level and prognosis of patients with glioma. Western blot and real-time PCR were used to detect UNR expression level in glioma cell lines and tissues. Next, UNR siRNAs were transfected in glioma cells, and MTS assay and scratch wound-healing assay were used to detect changes in cell proliferation and migration. Then, the candidate UNR target mRNAs were identified by analyzing the sequencing data of UNR iCLIP-seq, RNA sequencing and ribosome profiling databases of human melanoma. RNA immunoprecipitation and biotin pull-down assays were used to identify the UNR target mRNAs in glioma cells. Finally, western blot was used to detect the effect of UNR knockdown on ribosomal protein L9 (RPL9) and RPL9 protein expression level in glioma cell lines. RPL9 siRNA was transfected in A172 and T98G and the expression of vimentin in the cells was detected with western blot.Results Bioinformatics analysis showed that UNR mRNA expression level was significantly higher in high-grade glioma [Grade Ⅱ (n=126), Grade Ⅲ (n=51), Grade Ⅳ (n=128), P<0.001]. UNR high expression levels were associated with poor prognosis (P=0.0177). UNR had high expression level in glioma cell lines and patient samples compared with normal cell lines and normal brain samples (P<0.01). Knockdown of UNR inhibited glioma cells migration (P<0.05), but did not inhibit glioma cells growth in three glioma cell lines. UNR binded the 3’ untranslated region (UTR) of PTEN and RPL9 mRNAs. RPL9 protein was significantly highly expressed in most glioma cell lines (n=9) and knockdown of UNR resulted in a downregulation of RPL9 protein expression. Epithelial-mesenchymal transition (EMT)-related marker—vimentin was positively regulated by RPL9.Conclusions UNR could bind to the 3’UTR of PTEN and RPL9 in glioma cell lines, therefore promoting glioma cell migration and regulating the expression of RPL9. Here, we establish a link between UNR and RPL9 protein, which will provide new ideas for the further study of glioma.

Key words: UNR, glioma, migration, ribosomal protein L9, vimentin

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