Knockdown of NUSAP1 inhibits cell proliferation and invasion through downregulation of TOP2A in human glioblastoma

Cell Cycle. 2022 Sep;21(17):1842-1855. doi: 10.1080/15384101.2022.2074199. Epub 2022 May 13.

Abstract

Nucleolar and spindle associated protein 1 (NUSAP1), an indispensable mitotic regulator, has been reported to be involved in the development, progression, and metastasis of several types of cancer. Here, we investigated the expression and biological function of NUSAP1 in human glioblastoma (GBM), an aggressive brain tumor type with largely ineffective treatment options. Analysis of the molecular data in CGGA, TCGA and Rembrandt datasets demonstrated that NUSAP1 was significantly upregulated in GBM relative to low grade gliomas and non-neoplastic brain tissue samples. Kaplan-Meier analysis indicated that patients with tumors showing high NUSAP1 expression exhibited significantly poorer survival in both CGGA (P = 0.002) and Rembrandt cohorts (P = 0.017). Analysis of RNA sequencing data from P3-cells with stable knockdown of NUSAP1 revealed topoisomerase 2A (TOP2A) as a possible molecule downregulated by the loss of NUSAP1. Molecular analysis of the CGGA data revealed a strong correlation between NUSAP1 and TOP2A expression in primary gliomas and recurrent gliomas samples. SiRNA knockdown of either NUSAP1 or TOP2A in U251, T98 and GBM derived patient P3 cells inhibited GBM cell proliferation and invasion, and induced cell apoptosis. Finally, stable knockdown of NUSAP1 with shRNA led to decreased tumor growth in an orthotopic xenograft model of GBM in mice. Taken together, NUSAP1 gene silencing induced apoptosis possibly through the downregulation of the candidate downstream molecule TOP2A. Interference with the expression of NUSAP1 might therefore inhibit malignant progression in GBM, and NUSAP1 might thus serve as a promising molecular target for GBM treatment.

Keywords: GBM; NUSAP1; TOP2A; cell proliferation; invasion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain Neoplasms* / pathology
  • Cell Line, Tumor
  • Cell Proliferation
  • DNA Topoisomerases, Type II* / genetics
  • Down-Regulation / genetics
  • Gene Expression Regulation, Neoplastic
  • Glioblastoma* / pathology
  • Glioma* / genetics
  • Humans
  • Mice
  • Microtubule-Associated Proteins* / genetics
  • Poly-ADP-Ribose Binding Proteins* / genetics
  • RNA, Small Interfering / genetics

Substances

  • Microtubule-Associated Proteins
  • NUSAP1 protein, human
  • Poly-ADP-Ribose Binding Proteins
  • RNA, Small Interfering
  • DNA Topoisomerases, Type II
  • TOP2A protein, human

Grants and funding

This work was supported by the Natural Science Foundation of China (81972351, 81702475, 81803045 and 81702474), the Special Foundation for Taishan Scholars (tshw201502056 and tsqn201909173), the Department of Science & Technology of Shandong Province (2017CXGC1502), the China Postdoctoral Science Foundation (2018M642666, 2020T130371 and 2020M672072), the Jinan Science and Technology Bureau of Shandong Province (201704083 and 2019GXRC006), and the Shandong Research Institute of Industrial Technology.