Long Non-Coding RNA HAND2 Antisense RNA 1 Inhibits Colorectal Cancer Development by Sponging microRNA-3118/Leptin Receptor Axis

Ann Clin Lab Sci. 2023 Jan;53(1):82-93.

Abstract

Objective: This study aimed to examine the function of long non-coding RNA HAND2 antisense RNA 1 (HAND2-AS1) in colorectal cancer (CRC) and explore its underlying mechanism of action.

Methods: HAND2-AS1, microRNA (miR)-3118, and leptin receptor (LEPR) levels were determined using western blot analysis and reverse transcription quantitative polymerase chain reaction (RT-qPCR). RNA-binding protein immunoprecipitation (RIP) and luciferase reporter assays were performed to evaluate the relationship between HAND2-AS1, miR-3118, and LEPR. Overexpression of genes in CRC cell lines was performed by transfection with the overexpression vector or miR-mimic. Cell proliferation, migration, and apoptosis-related protein levels were evaluated using the Cell Counting Kit-8 (CCK-8), Transwell, and western blotting assays. A CRC xenograft mouse model was established to verify the role of HAND2-AS1 in CRC in vivo.

Results: In both CRC cell lines and CRC tumor samples the HAND2-AS1 expression was reduced. Upregulation of HAND2-AS1 levels inhibited CRC cell line proliferation and migration, initiated apoptosis, and suppressed the growth of CRC xenografted tumors. In addition, HAND2-AS1 sponges miR-3118, which is up-regulated in CRC. Moreover, miR-3118 overexpression promoted CRC cell line proliferation along with cell migration, but hindered apoptosis of cells, along with altering the consequences of high expression levels of HAND2-AS1 in CRC cells. In addition, miR-3118 can target LEPR, which is downregulated in CRC. The effect of miR-3118 on CRC cells was blocked by LERP overexpression.

Conclusion: HAND2-AS1 effectively inhibited CRC progression by sponging the miR-3118-LEPR axis. Our results may facilitate the development of therapeutic interventions for CRC.

Keywords: Colorectal cancer; LEPR; lncRNA HAND2-AS1; miR-3118.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Proliferation / genetics
  • Colorectal Neoplasms* / metabolism
  • Gene Expression Regulation, Neoplastic / genetics
  • Humans
  • Mice
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • RNA, Long Noncoding* / genetics
  • RNA, Long Noncoding* / metabolism
  • Receptors, Leptin / genetics
  • Receptors, Leptin / metabolism

Substances

  • MicroRNAs
  • RNA, Long Noncoding
  • Receptors, Leptin