Knockdown of long non-coding RNA GHET1 suppresses cervical carcinoma in vitro and in vivo

Cancer Biomark. 2020;28(1):21-32. doi: 10.3233/CBM-190269.

Abstract

Introduction: The present study evaluated the effects of the long non-coding RNA (lncRNA) gastric carcinoma high-expressed transcript (GHET1) in cervical carcinoma development.

Methods: The expression levels of GHET1 and PTEN were measured using in situ hybridisation, immunohistochemistry (IHC) and quantitative reverse transcription PCR assay to investigate their correlations. In an in vitro study, the effects of GHET1 knockdown on the biological activities of SiHa and HeLa cells were evaluated by MTT, flow cytometry, transwell and wound-healing assays and relative protein expression was measured using western blotting. In an in vivo experiment, cell apoptosis and relative protein expression were measured in nude mice using TUNEL and IHC assays, respectively.

Results: The expression levels of lncRNA GHET1 and PTEN protein differed significantly between cancer and adjacent normal tissues (P< 0.05) and were negatively correlated in the clinical data. In vitro, proliferation rateswere significantly down-regulated in SiHa and HeLa cells. The GHET1 knockdown (si-GHET1) groups showed significantly higher G1 phase and apoptosis rates and significantly suppressed invasion and migration abilities compared with the normal control (NC) group (P< 0.05 for all). The expression levels of PTEN, PI3 K, AKT, P53, matrix metalloproteinase (MMP)-2 and MMP-9 proteins differed significantly between the si-GHET1 and NC groups (P< 0.05 for all). In vitro, the lncRNA group showed significantly suppressed tumour volume and weight, increased cell apoptosis and different relative protein expression levels compared with the NC group (P< 0.05 for all).

Conclusion: GHET1 knockdown suppressed cervical carcinoma development via the PTEN/PI3 K/AKT signalling pathway.

Keywords: AKT; GHET1; HeLa; PI3 K; PTEN; SiHa.

MeSH terms

  • Adult
  • Animals
  • Cell Movement / physiology
  • Cell Proliferation / physiology
  • Female
  • Gene Knockdown Techniques
  • HeLa Cells
  • Heterografts
  • Humans
  • Immunohistochemistry
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • PTEN Phosphohydrolase / biosynthesis
  • PTEN Phosphohydrolase / genetics
  • PTEN Phosphohydrolase / metabolism
  • RNA, Long Noncoding / biosynthesis
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism*
  • Uterine Cervical Neoplasms / genetics*
  • Uterine Cervical Neoplasms / metabolism
  • Uterine Cervical Neoplasms / pathology

Substances

  • GHET1 transcript, human
  • RNA, Long Noncoding
  • PTEN Phosphohydrolase
  • PTEN protein, human