Characterisation of Levonorgestrel-Resistant Endometrial Cancer Cells

Endometrial cancer (EC) is the most common gynaecologic malignancy in the developed world, and incidence is increasing in premenopausal women. The levonorgestrel intrauterine system (LNG-IUS) is gaining traction as an alternative treatment for hyperplasia and early-stage EC for women who are unable to undergo surgery. Thirty to 60% of the women do not respond to this treatment, making the unknown mechanisms of levonorgestrel (LNG) resistance a critical obstacle for the conservative management of EC. This study aimed to characterise LNG-IUS treatment resistance in early-stage endometrial cancer in cell-line models.

LNG-resistant endometrial cancer cell lines (MFE296^R and MFE319^R) and cultures from three early stage endometrial cancer patients were developed. The behavioural profile of MFE296^R and MFE319^R was analysed using proliferation, adhesion, migration (wound healing and transwell) and invasion (spheroid) assays. LNG-sensitive cell lines (MFE296^S and MFE319^S) were compared to LNG^R cell lines (MFE296^R and MFE319^R). A literature search was conducted to identify possible candidate biomarkers of LNG resistance. RT-qPCR was used to analyse the mRNA expression of 17 candidate biomarkers in MFE296^R and MFE319^R. mRNA expression of the top differentially expressed genes was measured using RT-qPCR in primary cultures.

LNG resistance did not affect proliferation or invasion in immortalised endometrial cancer cells. Transwell migration was significantly increased in MFE319^R cells (p=0.03). Cellular adhesion significantly decreased in both MFE296^R cells (p=0.012) and MFE319^R cells (p=0.04). mRNA expression of KLF4 and SATB2 was significantly amplified in MFE296^R and MFE319^R cells. mRNA expression of KLF4 was significantly upregulated LNG-resistant primary cell lines.

LNG-resistant cells may have more oncogenic potential than their LNG-sensitive counterparts. Significant changes in the mRNA expression of KLF4 and SATB2 of LNG-resistant cells is a promising preliminary result in biomarker discovery for guiding LNG-IUS treatment of early stage endometrial cancer.