In Vitro Methods for Analyzing miRNA Roles in Cancer Cell Proliferation, Invasion, and Metastasis
MicroRNAs (miRNAs) are small noncoding, single-stranded RNAs consisting of 20–24 nucleotides (Bartel, Cell 116:281–297, 2004), which regulate target genes expression by interacting with 3′-untranslated regions (3′-UTRs) of target mRNAs, leading to translation repression or mRNA degradation (Filipowicz et al., Nat Rev Genet 9:102–114, 2008; Nilsen, Trends Genet 23:243–249, 2007). Accumulating evidence has elucidated them as oncogenes or tumor suppressors in cancers such as hepatocellular carcinoma, breast cancer, and lung cancer (Liu et al., Gastroenterology 136:683–693, 2009; Yu et al., Cell 131:1109–1123, 2007; Zhou et al., Sci Rep 7:42680, 2017; Iorio and Croce, Carcinogenesis 33:1126–1133, 2012). MiRNAs are involved in various biological processes, including cell proliferation (Liu et al., Mol Cancer Res 11:1314–1325, 2013), differentiation (Liu et al., Mol Cancer Res 11:1314–1325, 2013), apoptosis (Pan et al., Oncol Res 24:429–435, 2016), invasion and metastasis (Liu et al., Nat Commun 8:14270, 2017). Understanding the role of miRNAs in tumor gives new perspective on cancer diagnosis and therapy (Rupaimoole and Slack, Nat Rev Drug Discov 16:203–222, 2017; Berindan-Neagoe et al., CA Cancer J Clin 64:311–336, 2014). This chapter will focus on the in vitro methods for identifying miRNAs roles in cell proliferation, invasion, and metastasis in tumor development, which includes CCK-8 assay, Wound Healing assay, and Transwell assay.
Key wordsMiRNA Proliferation Invasion Metastasis
This study was supported by the Key Laboratory of Laboratory Medicine of Jiangsu Province of China and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions. We are grateful to the technical support from the National Key Clinical Department of Laboratory Medicine of Jiangsu Province Hospital.