The Joint Annual Scientific Meetings of the Endocrine Society of Australia and the Society for Reproductive Biology 2018

miR-29c overexpression in endometrial cancer reduced proliferation and increased apoptosis and altered cell cycle and migration related genes (#61)

Michelle Van Sinderen 1 2 , Meaghan Griffiths 1 2 3 , Keith Niven 4 , Evdokia Dimitriadis 1 2 3
  1. Department of Molecular and Translational Medicine, Monash University, Clayton, vic, Australia
  2. Hudson Institute of Medical Research, Clayton, VIC, Australia
  3. Anatomy and Development, Monash University, Clayton, Victoria
  4. FlowCore, Technology Research Platforms, Monash University, Clayton, Victoria, Australia

Endometrial cancer is the most common gynaecological cancer and advanced disease has a very poor prognosis. There are also no biomarkers for endometrial cancer. Micro RNA (miRs) are known to be dysregulated in endometrial cancer. The expression of miR-29 family members (-a and –b) are downregulated in endometrial cancer.  However, a role for miR-29c and its targets in endometrial cancer growth remain to be elucidated.  The aim of this study was to investigate the functional effects of re-introducing miR-29c expression in endometrial cancer cell lines and identify which miR-29c targets may contribute to cancer progression.

Real-time PCR for miR-29c expression was performed on human endometrial tumour grades 1-3 and benign tissue as well as endometrial cancer cell lines Ishikawa, HEC1A and AN3CA.   Cell lines were transfected with miR-29c mimic and negative control.  xCELLigence real-time cell monitoring was performed to analyse proliferation and migration and Flow cytometry for apoptosis and cell cycle population percentages.  miR-29c target genes in the 3 cell lines were analysed using real-time PCR. 

miR-29c was down regulated in grade 1-3 endometrial cancer samples compared to benign endometrium (p<0.0001 respectively).  miR-29c expression was reduced in Ishikawa (p<0.0001) and AN3CA (p<0.01) but not HEC1A cell lines compared to non-cancerous primary human endometrial epithelial cells. Overexpression of miR-29c reduced proliferation (p<0.05 HEC1A and p<0.001 AN3CA at 72hr), increased apoptosis (p<0.001 Ishikawa and p<0.05 AN3CA) and reduced miR-29c target genes (CAV1, CDC42, SIRT1, FBN1, HBP1, ITGB1, NUMB, MCL1, MCL1, SGK1, VEGFA and MDM2) across the three cell lines. Inhibition of miR-29c in HEC1A cells increased proliferation (p<0.05 at 72hr) and COL4A1 expression (p<0.01).

The re-introduction of miR-29c to endometrial cancer cell lines reduced proliferation, increased apoptosis and reduced miR-29c target genes in vitro. This suggests miR-29c may be a therapeutic target for endometrial cancer.