High miR-100 expression is associated with aggressive features and modulates TORC1 complex activation in lung carcinoids

// Ida Rapa 1 , Arianna Votta 1 , Gaia Gatti 1 , Stefania Izzo 1 , Nicola Lo Buono 2 , Elisa Giorgio 2 , Simona Vatrano 1 , Francesca Napoli 1 , Aldo Scarpa 3 , Giorgio Scagliotti 1 , Mauro Papotti 1 and Marco Volante 1 1 Department of Oncology at San Luigi Hospital, University of Turin, Turin, Italy 2 Department of Medical Sciences, University of Turin, Turin, Italy 3 ARC-NET Applied Research on Cancer Centre at Department of Diagnostics and Public Health, Section of Pathology, University and Hospital Trust of Verona, Verona, Italy Correspondence to: Marco Volante, email: marco.volante@unito.it Keywords: miRNAs; miR-100; lung carcinoids; rapamycin sensitivity; mTOR Received: November 09, 2017 Accepted: May 14, 2018 Published: June 08, 2018 ABSTRACT Purpose: Mammalian target of rapamycin (mTOR) is a promising therapeutic target in advanced lung carcinoid patients. However, the mechanisms of mTOR modulation and of responsiveness to mTOR inhibitors are largely unclear. Our aim was to analyze the expression and functional role of specific miRNAs in lung carcinoids as an alternative mechanism targeting mTOR pathway. Experimental design: Seven miRNAs, selected by bioinformatic tools and literature search, were analyzed in 142 lung neuroendocrine neoplasms (92 carcinoids and a control group of 50 high grade neuroendocrine carcinomas), and compared with mTOR mRNA expression and clinical/pathological parameters. Tissue results were validated in vitro in two lung carcinoid cell lines by specific RNA interference and biological/pharmacological tests. Results: Tissutal expression of five miRNAs (miR-99b, miR-100, miR-155, miR-193a-3p, miR-193a-5p) was inversely correlated with mTOR mRNA expression, supporting their role in the negative regulation of mTOR transcription. High expression of miR-100, miR-193a-3p and miR-193a-5p was associated with aggressive features and, for the former two, with shorter time to progression. In H727 and UMC11 lung carcinoid cells, miR-100 modulated mTOR RNA and TORC1 complex protein expression, positively promoted cell migration and negatively influenced cell proliferation. Moreover, miR-100 directly influenced responsiveness of H727 and UMC11 cells to rapamycin. Conclusions: MiR-100 actively participates to the regulation of mTOR expression in lung carcinoids and represents a novel candidate prognostic biomarker for this tumor type; moreover, inhibition of its expression is associated to increased responsiveness to mTOR inhibitors and might represent a novel strategy to sensitize lung carcinoids to these target agents.