Overexpression of IGF2BP3 As a Potential Oncogene in Ovarian Clear Cell Carcinoma

Abstract

Ovarian Clear Cell Carcinoma (OCCC) displays distinctive clinical and molecular characteristics and confers the worst prognosis among all ovarian carcinoma histotypes when diagnosed at advanced stage because of the lack of effective therapy. IGF2BP3 is an RNA binding protein that modulates gene expression by posttranscriptional action. In this study, we investigated the roles of IGF2BP3 in the progression of OCCC. We used 328 OCCCs from the AOVT (the Alberta Ovarian Tumor Type study) and the COEUR (the Canadian Ovarian Experimental Unified Resource) cohorts to elucidate the associations between IGF2BP3 expression and clinicopathological parameters, with positive IGF2BP3 expression defined as diffuse block staining, being more frequently observed at stage III (P=0.0056) and significantly associated with unfavorable overall survival (HR=1.59, 95% CI 1.09 - 2.33) in multivariate analysis. IGF2BP3 mRNA gene expression was markedly increased in OCCC cell lines compared to normal tissue such as ovarian surface epithelium. We chose two IGF2BP3-overexpressing cell lines ES2 and OVMANA for in vitro and in vivo knockdown experiments. The proliferation and viability of both cell lines were significantly inhibited by 2 IGF2BP3 siRNAs and similar suppression was observed in cell migration and invasion by Wound Healing and Transwell assays. The percentage of apoptotic cancer cells was enhanced by both IGF2BP3 siRNAs. In vivo experiments showed a significantly reduced size of tumors when treated with IGF2BP3 siRNA compared to controls. Furthermore, cancer metastasis-indicators MMP2 and MMP9 proteins were down-regulated. In conclusion, our study shows that IGF2BP3 expression is a promising biomarker for prognostication of women diagnosed with OCCC with multiple effects on key cell functions, supporting its role as an important cellular regulator with potential oncogenic activity, and as a potential target for future intervention strategies. Funding Statement: This study used resources provided by the Canadian Ovarian Cancer Research Consortium’s COEUR biobank funded by the Terry Fox Research Institute (grant #2009-15) and supervised by the Centre hospitalier de l’Universite de Montreal (CHUM). The Consortium acknowledges contributions to its COEUR biobank from institutions across Canada. Wenrang Kan participated in basic experimental work as a volunteer. This work was supported by grants of the National Natural Science Foundation of China (NSFC30970119, 81030029, 81271786, NSFC-NIH 81161120416, 81671980, 81871623) and College students’ Innovation & Entrepreneurship project in Heilongjiang Province (201410226047 J.J.K, D.Y.; 201510226020 D.S.L, L.Y., T.L., L.Q., L.L.G.; 201610226095 H.Y.W., Z.H.S. T.T.G., S.J.H., S.G.; 201610226094 Y.Y.Q., M.Y.; 201710226073 S.J.H., S.G.). H.D.L. is supported by a scholarship from China Scholarship Council, CSC No. 201508230143, for an academic visit to the University of Calgary (Univ. of Calgary ID number: 30016355). We also thank the Health and Family Planning Commission of Heilongjiang Province (2016-188), the Fundamental Research Funds for the Provincial Universities (2017JCZX57), the Young Innovative Talents in Universities of Heilongjiang Province (UNPYSCT-2018064), the China Postdoctoral Science Foundation (2018M630380) and the Heilongjiang Postdoctoral Financial Assistance (LBH-Z18198) program for their support. Declaration of Interests: There are no competing interest concerns with any of the authors. Ethics Approval Statement: All animal experiments were executed in accordance with the Institutional Ethics Committee of Harbin Medical University.