Silencing NID2 by DNA Hypermethylation Promotes Lung Cancer

Pathol Oncol Res. 2020 Apr;26(2):801-811. doi: 10.1007/s12253-019-00609-0. Epub 2019 Mar 2.

Abstract

To characterize the DNA methylation as well as exploring the relationship between NID2 methylation and the lung cancer development. Collecting chip data of 9 lung cancer samples and 11 adjacent normal samples from the Gene Expression Omnibus database. Tissues and cells NID2 gene methylation level was measured by methylation-specific PCR. NID2 mRNA level and protein level were validated by Real-Time PCR and Western blot separately. Functional study of lung cancer cells was performed with Cell Counting Kit-8 assay. Colony formation assay, transwell assay, wound healing assay and low cytometry were performed. Finally, NID2 tumorigenesis in vivo was tested in nude mice xenograft models. Microarray analysis outcome present NID2 hypermethylation status in lung cancer tissues. High methylation and low mRNA expression levels of NID2 were detected. After NID2 demethylation or overexpression in cancer cells, cell viability, proliferation, migration as well as invasion ability decreased. Nevertheless, a significant enhancement in apoptosis rate were observed. Overexpressing NID2 or demethylation in lung cancer cells inhibited the tumorigenesis of lung cancer in nude mice. The mRNA and protein level of NID2 in tumors obtained from nude mice xenograft were unanimous with the in vitro assays' outcome, which significantly decreased after overexpressing NID2 or demethylation. NID2 methylation reduces its expression level and promotes the development of lung cancer.

Keywords: DNA methylation; Lung cancer; NID2; Nude mice xenograft.

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Calcium-Binding Proteins / genetics*
  • Carcinogenesis / genetics
  • Cell Adhesion Molecules / genetics*
  • Cell Proliferation / genetics
  • DNA Methylation / genetics*
  • Female
  • Gene Silencing
  • Heterografts
  • Humans
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / pathology*
  • Male
  • Mice
  • Mice, Nude

Substances

  • Calcium-Binding Proteins
  • Cell Adhesion Molecules
  • NID2 protein, human