Background: Small RNAs play an essential role in fundamental biological processes such as differentiation, proliferation, apoptosis and homeostasis. Evidence suggests that small RNAs may also play function in Cancer progression. The conventional in-vitro two-dimensional (2D) monolayer cell culture models are not sufficient to explain the exact mechanism behind tumor invasion and metastasis. The objective of this study is to analyze the role of small RNAs in tumor invasion and metastasis using in vitro 2D and anchorage independent (3D) models in ovarian cancer cell lines. Methods: Total RNA samples were isolated from several 2D and 3D ovarian cancer cell lines. Small RNA libraries were prepared and sequenced using Next-Generation Sequencing. Results: Gene expression levels of 3D culture differ from 2D culture, especially in epithelial and mesenchymal target genes. We were able to identify the predominant pathways that small RNAs regulate in 3D cultures. These pathways were involved in cellular invasion, migration, metastasis, proliferation, and tumor growth. Conclusion: The Results from this experiment suggest that small RNAs play a crucial role in controlling tumor invasion and metastasis in the studied phenotypes. Therefore, small RNAs may offer a critical target for the development of anti-cancer drugs.


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