Upregulation of miR-129-5p affects laryngeal cancer cell proliferation, invasiveness, and migration by affecting STAT3 expression

Tumour Biol. 2016 Feb;37(2):1789-96. doi: 10.1007/s13277-015-3969-y. Epub 2015 Aug 29.

Abstract

microRNA-129-5p may have a relationship with laryngeal squamous cell carcinoma (LSCC), but the expression and function of miRNA-129-5p are not fully understood. This study was performed to investigate the expression and function of miRNA-129-5p in LSCC and its effects on STAT3. HBE and Hep-2 cells were cultured and analyzed by western blotting and real-time PCR. miR-129-5p expression was detected by real-time PCR. Hep-2 cells were transfected with a miR-129-5p expression vector. Cell proliferation, invasion, migration, cell cycle, and apoptosis assays were used to determine the role of miRNA-129-5p in the growth, invasion, and migration of LSCC cells and to determine the importance of STAT3 in these effects. STAT3 mRNA expression in Hep-2 cells was significantly higher than that in HBE cells (P < 0.05). miR-129-5p expression detected by real-time PCR showed that it was expressed at a lower level in Hep-2 cells than in HBE cells (P < 0.05). Compared with the control cells, the transfected cells showed lower STAT3 mRNA expression. For up to 7 days in culture, the transfected cells showed lower proliferation than the control cells (P < 0.05). After 24 h in culture, the apoptosis rate in miR-129-5p-transfected cells was 3.48 ± 0.38 %, while the rate in control cells was 0.92 ± 0.09 % (P = 0.0028), but the statistical significance was lost after 72 h in culture (P = 0.3180). The invasion and migration of the cells were inhibited after 24 and 72 h in culture when the miR-129-5p expression in Hep-2 cells was upregulated (P = 0.0037 and 0.00383, respectively), while there was no statistically significant difference at 48 h (P = 0.0712). STAT3 expression could be suppressed by the upregulation of miR-129-5p expression. Both the proliferation and migration of tumor cells were suppressed when the level of STAT3 expression was decreased. The apoptosis rate of tumor cells was also increased. Based on these data, we suggest that miR-129-5p may directly inhibit STAT3 expression and play an important role in the development of LSCC.

Keywords: Apoptosis; Invasion; Laryngeal squamous cells carcinoma; STAT3; miRNA-129-5p.

MeSH terms

  • Apoptosis / genetics
  • Carcinoma, Squamous Cell / genetics
  • Carcinoma, Squamous Cell / pathology
  • Cell Cycle / genetics
  • Cell Line, Tumor
  • Cell Movement / genetics*
  • Cell Proliferation / genetics*
  • Gene Expression Regulation, Neoplastic / genetics
  • Humans
  • Laryngeal Neoplasms / genetics*
  • Laryngeal Neoplasms / pathology
  • MicroRNAs / genetics*
  • Neoplasm Invasiveness / genetics*
  • Neoplasm Invasiveness / pathology
  • RNA, Messenger / genetics
  • STAT3 Transcription Factor / genetics*
  • Transfection / methods
  • Up-Regulation / genetics*

Substances

  • MicroRNAs
  • Mirn129 microRNA, human
  • RNA, Messenger
  • STAT3 Transcription Factor
  • STAT3 protein, human