Function of miR-10a Overexpression in Ovarian Clear Cell Carcinoma
Davis, Noah, Indiana University School of Medicine; Kaitlyn Collins, Indiana University School of Medicine Department of Biochemistry and Molecular Biology; Shannon Hawkins, Indiana University School of Medicine Department of Obstetrics and Gynecology
Background/Significance/Rationale: Ovarian cancer is the most lethal cancer of the female reproductive tract in the United States, responsible for the death of 14,000 US women each year. Ovarian clear cell carcinoma (OCCC) has higher rates of chemoresistance, and lower overall survival compared to women with high grade serous ovarian carcinoma. Previous RNA sequencing of human OCCC samples in the Hawkins Lab, found an 11- fold upregulation of miR-10a in OCCC with concurrent endometriosis compared to benign endometrioma. MiR-10a overexpression has been correlated with chemoresistance in cervical, breast, and lung cancers. We hypothesize that miR-10a drives progression of OCCC by dysregulating genes associated with cell proliferation and migration.
Methods: OVISE, SMOV2, OVTOKO, ES2, IGROV1, and RMG1 ovarian clear cell carcinoma cell lines were utilized to evaluate the role of miR-10a on cell proliferation and gene expression. In vitro transfection of miR-10a mimic, miR-10a inhibitor, and negative control was performed. Cell proliferation was visualized using MTS assay. Gene expression analysis was performed by qPCR. Varying concentrations of miR-10a mimic – 20nM, 50nM, and 100nM – were transfected to evaluate dose response in potential miR-10a target genes: CDK6, CHL1, EPHA4, SERPINE1, and NFASC.
Results/Findings: The presence of miR-10a variably influenced cell proliferation. However, in the SMOV2 cell line, miR-10a mimic significantly decreased cell proliferation. Increased miR-10a concentrations may be associated with decreased gene expression of select genes in the OVISE cell line.
Conclusions/Discussion: Our findings provide some insight into the functions of miR-10a, though more investigation is warranted. Future work will investigate the relationship between varying miR-10a mimic concentrations, gene expression, and cell growth. Cell cycle analysis will also be completed to determine cell-cycle dependent functions of miR-10a.
Translational/Human Health Impact: Understanding miR-10a function in OCCC may allow miR-10a to be utilized as a biomarker to detect cancer progression or indicate levels of chemoresistance.