LncRNA DANCR affected cell growth, EMT and angiogenesis by sponging miR-345-5p through modulating Twist1 in cholangiocarcinoma

Eur Rev Med Pharmacol Sci. 2020 Mar;24(5):2321-2334. doi: 10.26355/eurrev_202003_20498.

Abstract

Objective: LncRNA DANCR has been reported to play an important role in various cancers. Therefore, this study aimed at exploring the function and regulatory mechanism of DANCR in Cholangiocarcinoma (CCA).

Patients and methods: qRT-PCR was used to measure the expression of DANCR, miR-345-5p in tissues and cells. Western blot was applied to measure the protein expression of Twist, N-cadherin, Vimentin, E-cadherin, VEGF-A, VEGF-C, PCNA and C-caspase 3. The relationship between DANCR and miR-345-5p was determined by luciferase reporter assay. MTT assay and flow cytometry were used to assess cell proliferation and apoptosis, respectively. Transwell assay was performed to detect cell invasion and migration.

Results: We found that the expression of DANCR was significantly induced in CCA tissues and cells. Inhibition of DANCR remarkably suppressed CCA cell proliferation, migration, invasion, EMT and angiogenesis as well as induced cell apoptosis in vitro and in vivo. Luciferase reporter assay determined that DANCR directly targeted miR-345-5p and Twist1 was a target mRNA of miR-345-5p. Otherwise, miR-345-5p down-expression partially reversed the effect induced by the suppression of DANCR in CCA. Moreover, the suppressive effects of high miR-345-5p expression on CCA cells were reversed by improving Twist1 expression.

Conclusions: In this study, we verified that LncRNA DANCR affected cell proliferation, migration, invasion, angiogenesis, epithelial-mesenchymal transition (EMT) and induced apoptosis through modulating miR-345-5p/Twist1 axis in Cholangiocarcinoma.

MeSH terms

  • Animals
  • Apoptosis
  • Bile Duct Neoplasms / metabolism*
  • Bile Duct Neoplasms / pathology
  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Cholangiocarcinoma / metabolism*
  • Cholangiocarcinoma / pathology
  • Epithelial-Mesenchymal Transition / genetics
  • Female
  • Humans
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Middle Aged
  • Neovascularization, Pathologic / metabolism*
  • Neovascularization, Pathologic / pathology
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism*
  • Twist-Related Protein 1 / genetics
  • Twist-Related Protein 1 / metabolism*

Substances

  • DANCR long noncoding RNA, human
  • MIRN345 microRNA, human
  • MicroRNAs
  • Nuclear Proteins
  • RNA, Long Noncoding
  • TWIST1 protein, human
  • Twist-Related Protein 1