engleski

Downregulation of Hedgehog-GLI signaling pathway genes by miRNA molecules in high-grade serous ovarian carcinoma

Ovarian carcinoma is the deadliest of all gynecologic cancers. Its high death rate is a result of the fact that >60% patients are diagnosed in advanced stage. Hedgehog-GLI signaling pathway (HH-GLI) is involved in embryonal ovariogenesis but its atypical activity can lead to different types of ovarian tumors. Aberrant HH-GLI activity was observed in some ovarian tumor types previously, molecular causes of which are not jet fully known. We hypothesize that changes in the expression of miRNA molecules related to HH-GLI genes could contribute to the development of high-grade serous ovarian carcinoma (HGSOC), the most malignant and most difficult to detect at an earlier stage. Gene and miRNA expression profiling of 16 HGSOC and 8 healthy fallopian tube tissue samples was performed using Agilent microarray platforms. Data was analyzed using R/Bioconductor packages. Four tools were used to predict target genes of differentially expressed miRNAs. Expression of selected miRNAs and genes was verified using qPCR on an extended set of 10 fallopian tube and 55 HGSOC samples. Various cell-based assays were used to examine biological functions of discovered miRNA/mRNA pairs. Analysis gave a list of 55 miRNAs and 2782 genes differentially expressed in HGSOC. By comparing a list of predicted target genes and differentially expressed HH-GLI genes we revealed several potential miRNA/mRNA interactions. Co-expression of the following combinations has been verified: GLI3/miR-200b-3p ; BCL2/miR-96-5p and miR-21-5p, and IHH/miR-107 and miR-16-5p. Luciferase assays proved physical interactions of miRNAs and GLI3 and IHH 3’UTRs, and their impact on ovarian carcinoma cell biology was identified. We discovered several new miRNAs targeting HH-GLI genes and further studies would verify their usefulness as potential diagnostic and prognostic markers or even therapeutic targets for HGSOC. This study was funded by Terry Fox Foundation, City of Zagreb and Croatian Science Foundation (IP-2016-06-1268).

Hedgehog-GLI ; ovarian cancer ; microRNA ; mRNA ; expression

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