2. Academic Validation
  3. Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

  • J Exp Clin Cancer Res. 2022 Jan 24;41(1):33. doi: 10.1186/s13046-021-02230-z.
Hongyan Li 1 2 3 4 5 6,  Hong Zhang 7 8 9 10 11 12,  Guomin Huang 1 3 4 5 6 13,  Zhitong Bing 5 6 14,  Duling Xu 1 3 4 5 6 13,  Jiadi Liu 1 3 4 5 6 13,  Hongtao Luo 15 16,  Xiaoli An 14
Affiliations

Affiliations

  • 1 Department of Medical Physics, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000, China.
  • 2 Gansu Provincial Isotope Laboratory, Lanzhou, 730300, China.
  • 3 Key Laboratory of Heavy Ion Radiation Biology and Medicine of Chinese Academy of Sciences, Lanzhou, 730000, China.
  • 4 Key Laboratory of Basic Research on Heavy Ion Radiation Application in Medicine, Lanzhou, 730000, China.
  • 5 School of Nuclear Science and Technology, University of Chinese Academy of Sciences, Beijing, 101408, China.
  • 6 Advanced Energy Science and Technology Guangdong Laboratory, Huizhou, 516029, China.
  • 7 Department of Medical Physics, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000, China. [email protected].
  • 8 Gansu Provincial Isotope Laboratory, Lanzhou, 730300, China. [email protected].
  • 9 Key Laboratory of Heavy Ion Radiation Biology and Medicine of Chinese Academy of Sciences, Lanzhou, 730000, China. [email protected].
  • 10 Key Laboratory of Basic Research on Heavy Ion Radiation Application in Medicine, Lanzhou, 730000, China. [email protected].
  • 11 School of Nuclear Science and Technology, University of Chinese Academy of Sciences, Beijing, 101408, China. [email protected].
  • 12 Advanced Energy Science and Technology Guangdong Laboratory, Huizhou, 516029, China. [email protected].
  • 13 University of Chinese Academy of Sciences, Beijing, 100039, China.
  • 14 Department of Computational Physics, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000, China.
  • 15 Department of Radiation Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, 730000, China.
  • 16 Radiation Oncology of Gansu Provincial Cancer Hospital, Lanzhou, 730030, China.
PMID: 35073964 DOI: 10.1186/s13046-021-02230-z
Abstract

Background: Depletion of certain ribosomal proteins induces p53 activation, which is mediated mainly by ribosomal protein L5 (RPL5) and/or ribosomal protein L11 (RPL11). Therefore, RPL5 and RPL11 may link RPs and p53 activation. Thus, this study aimed to explore whether RPs interact with RPL11 and regulate p53 activation in lung adenocarcinoma (LUAD) cells.

Methods: The endogenous RPL11-binding proteins in A549 cells were pulled down through immunoprecipitation and identified with a proteomics approach. Docking analysis and GST-fusion protein assays were used to analyze the interaction of ribosomal protein S27a (RPS27a) and RPL11. Co-immunoprecipitation and in vitro ubiquitination assays were used to detect the effects of knockdown of RPS27a on the interaction between RPS27a and RPL11, and on p53 accumulation. Cell cycle, Apoptosis, cell invasion and migration, cell viability and colony-formation assays were performed in the presence of knockdown of RPS27a. The RPS27a mRNA expression in LUAD was analyzed on the basis of the TCGA dataset, and RPS27a expression was detected through immunohistochemistry in LUAD samples. Finally, RPS27a and p53 expression was analyzed through immunohistochemistry in A549 cell xenografts with knockdown of RPS27a.

Results: RPS27a was identified as a novel RPL11 binding protein. GST pull-down assays revealed that RPS27a directly bound RPL11. Knockdown of RPS27a weakened the interaction between RPS27a and RPL11, but enhanced the binding of RPL11 and murine double minute 2 (MDM2), thereby inhibiting the ubiquitination and degradation of p53 by MDM2. Knockdown of RPS27a stabilized p53 in an RPL11-dependent manner and induced cell viability inhibition, cell cycle arrest and Apoptosis in a p53-dependent manner in A549 cells. The expression of RPS27a was upregulated in LUAD and correlated with LUAD progression and poorer prognosis. Overexpression of RPS27a correlated with upregulation of p53, MDM2 and RPL11 in LUAD clinical specimens. Knockdown of RPS27a increased p53 activation, thus, suppressing the formation of A549 cell xenografts in nude mice.

Conclusions: RPS27a interacts with RPL11, and RPS27a knockdown enhanced the binding of RPL11 and MDM2, thereby inhibiting MDM2-mediated p53 ubiquitination and degradation; in addition, RPS27a as important roles in LUAD progression and prognosis, and may be a therapeutic target for patients with LUAD.

Keywords

Apoptosis; Lung adenocarcinoma; Ribosomal protein S27a (RPS27a).

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