Epigenetic silencing of Rab39a promotes epithelial to mesenchymal transition of cervical cancer through AKT signaling

Exp Cell Res. 2019 May 15;378(2):139-148. doi: 10.1016/j.yexcr.2019.02.025. Epub 2019 Feb 28.

Abstract

The objective of this study was to investigate the functional role of Rab39a in human cervical cancer (CC) and the underlying molecular mechanisms. We first measured Rab39a mRNA expression in CC tissues and paired non-tumor tissues by quantitative real-time PCR (QRT-PCR). Overall survival of CC patients with different mRNA levels of Rab39a in The Cancer Genome Atlas (TCGA) database was assessed by Kaplan-Meier survival curves analysis. Next methylation-specific PCR (MSP) was performed to determine the expression mechanism of Rab39a. Then cell proliferation, migration and invasion of Rab39a-transfected or mock-transfected cervical cancer cells were determined by CCK-8, flow cytometry, wound healing, transwell migration and invasion assays, respectively. Finally, the molecular mechanism by which Rab39a modulated CC cell epithelial-mesenchymal transition (EMT) was explored. It was found that Rab39a mRNA was significantly down-regulated in the high-risk patients compared to the low-risk patients (p = 0.0054). Six of seven cancer tissues with lymph node metastasis express low Rab39a mRNA compared to the surrounding non-tumor tissues. Cervical cancer patients with low level of Rab39a were showed a poorly clinical outcome (p = 0.004). Loss of Rab39a expression in cervical cancer tissues was associated with the aberrant DNA methylation in the promoter of Rab39a gene. Disrupted Rab39a expression in cervical cancer cells could be restored after treatment with the demethylated agent 5-Aza-2'-deoxycytidine. Furthermore, it was found that Rab39a hardly influenced cell growth but significantly suppressed cell migration, invasion and EMT process. Rab39a exerted its potential suppressor functions through inhibiting AKT phosphorylation. The inhibition effects of Rab39a could be blocked by AKT pathway inhibitor. Collectively, our data shows that Rab39a is a potential epigenetic silenced tumor suppressor inhibiting cancer invasion and migration through modulating the AKT signaling.

Keywords: AKT signaling; DNA methylation; EMT; Rab39a.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Movement
  • Down-Regulation
  • Epigenesis, Genetic*
  • Epithelial-Mesenchymal Transition / genetics*
  • Female
  • Gene Expression Regulation, Neoplastic
  • Gene Silencing*
  • Humans
  • Methylation
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Signal Transduction*
  • Tumor Suppressor Proteins / genetics*
  • Tumor Suppressor Proteins / metabolism
  • Uterine Cervical Neoplasms / genetics*
  • Uterine Cervical Neoplasms / metabolism
  • rab GTP-Binding Proteins / genetics*
  • rab GTP-Binding Proteins / metabolism

Substances

  • Tumor Suppressor Proteins
  • Proto-Oncogene Proteins c-akt
  • RAB39A protein, human
  • rab GTP-Binding Proteins