Knockdown of long non-coding RNA LINC00941 suppressed cell proliferation, colony-formation, and migration of human glioblastoma cell lines

Folia Neuropathol. 2023;61(2):209-216. doi: 10.5114/fn.2023.126894.

Abstract

Introduction: Glioblastoma (GBM) represents the most common and lethal type of primary brain tumour in adults, and due to its high invasiveness, treatment of GBM remains challenging. This work is aimed to elucidate the role of LINC00941 in GBM.

Material and methods: Expression of LINC00941 in two GBM cell lines U251 and U87-MG was knocked down using siRNA. Cell proliferation and colony-formation ability of LINC00941 knockdown were examined. Apoptosis of the knockdown was evaluated using flow cytometry, with the levels of Bax, Bcl-2, cleaved caspase-3, and phosphorylation of ERK and Akt to be examined using western blotting. Migration and invasion of the knockdown was studied using transwell assays.

Results: Expression of LINC00941 was significantly elevated in GBM compared to non-tumour tissues ( p < 0.01). Statistical analysis on the expression data further revealed the negative correlation between LINC00941 and miR-526b-5p ( r = 0.7494, p < 0.001). LINC00941 was successfully knocked down with RNA interference in U251 and U87-MG. The knockdown significantly suppressed cell proliferation and the ability to form colonies. Percentage of apoptotic cells was elevated by the knockdown in both cell lines as evidenced by flow cytometric analysis, which was accompanied by a significant decrease in Bcl-2 and substantial increases in Bax and cleaved caspase-3. Phosphorylation of ERK and Akt was also enhanced in both cell lines by the knockdown. In addition, knockdown of LINC00941 suppressed migration of both cell lines across transwell membrane and matrigel.

Conclusions: LINC00941 is overexpressed in GBM, exhibiting important roles in cell proliferation and survival, migration and invasion.

Keywords: LINC00941; glioblastoma; miR-526b-5p; long non-coding RNA.

MeSH terms

  • Adult
  • Caspase 3
  • Cell Line
  • Cell Proliferation / genetics
  • Glioblastoma* / genetics
  • Humans
  • Proto-Oncogene Proteins c-akt
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Long Noncoding* / genetics
  • bcl-2-Associated X Protein

Substances

  • Caspase 3
  • RNA, Long Noncoding
  • Proto-Oncogene Proteins c-akt
  • bcl-2-Associated X Protein
  • Proto-Oncogene Proteins c-bcl-2