Identification and Functional Validation of Differentially Expressed microRNAs in Ascites-Derived Ovarian Cancer Cells Compared with Primary Tumour Tissue

• Main Authors: Jiang Y, Shi Y, Lyu T, Liu H, Shen L, Zhou T, Feng W
• Format: Article
• Language: English
• Published: Dove Medical Press 2021-08-01
• Series: Cancer Management and Research
• Subjects: ovarian cancer; metastasis; ascites-derived spheroids; primary tumour tissue; micrornas
• Online Access: https://www.dovepress.com/identification-and-functional-validation-of-differentially-expressed-m-peer-reviewed-fulltext-article-CMAR

Yahui Jiang,* Yiwen Shi,* Tianjiao Lyu, Hua Liu, Lifei Shen, Tianyu Zhou, Weiwei Feng Department of Gynecology and Obstetrics, Ruijin Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Weiwei FengDepartment of Gynecology and Obstetrics, Ruijin Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai, 200025, People’s Republic of ChinaTel/Fax [phone]Email [e-mail]: Ovarian cancer, manifested by malignant ascites, is the most lethal gynaecological cancer. Suspended ascites-derived spheroids may contribute to ovarian cancer metastasis. MicroRNAs (miRNAs) are also associated with ovarian cancer metastasis. Here, we aimed to investigate the differentially expressed miRNAs (DE-miRNAs) in ascites-derived spheroids compared with primary tumour tissues, which may regulate ovarian cancer metastasis.Methods: The DE-miRNAs between ovarian cancer primary tumour tissues and ascites-derived spheroids were identified by GEO2R screening in samples from 3 high-grade serous ovarian cancer (HGSOC) patients of dataset GSE65819. We used MiRTarBase, TargetScanHuman7.2 and STRING to predict the target hub genes of DE-miRNAs and DAVID to perform functional analysis of hub genes. ALGGEN PROMO and TransmiR v2.0 were used to predict transcription factors (TFs) that potentially regulate DE-miRNAs expression. The observed differences in DE-miRNAs expression were validated with samples from 12 HGSOC patients and 2 ovarian cancer cell lines using PCR. The functions of DE-miRNAs on ovarian cancer progression were verified by invasion, adherent, and angiogenesis assays.Results: Through bioinformatics screening and experimental validation, miR-199a-3p, miR-199b-3p, miR-199a-5p, miR-126-3p and miR-145-5p were identified as being significantly downregulated in ascites-derived spheroids compared with primary tumour tissues. In addition, TFAP2A was identified as a potentially common upstream TF regulating the expression of the above mentioned DE-miRNAs. The overexpression of miR-199a-3p, miR-199b-3p, miR-199a-5p lead to invasion inhibition, and the overexpression of miR-126-3p, miR-145-5p, miR-199a-5p and miR-199b-3p lead to adhesion inhibition of suspended ovarian cancer cells. High-expressed miR-126-3p, miR-199a-3p, miR-199a-5p and miR-199b-3p contributed to apoptosis of suspended ovarian cancer cells.Conclusion: The downregulated expression of miR-199a-3p, miR-199b-3p, miR-199a-5p, miR-126-3p and miR-145-5p in ascites-derived spheroids plays a key role in promoting ovarian cancer progression, which may represent novel molecules for targeted therapy for ovarian cancer.Keywords: ovarian cancer, metastasis, ascites-derived spheroids, primary tumour tissue, microRNAs