JOURNAL ARTICLE

The functional characterization of long noncoding RNA SPRY4-IT1 in human melanoma cells

Joseph Mazar, Wei Zhao, Ahmad M Khalil, Bongyong Lee, John Shelley, Subramaniam S Govindarajan, Fumiko Yamamoto, Maya Ratnam, Muhammad Nauman Aftab, Sheila Collins, Brian N Finck, Xianlin Han, John S Mattick, Marcel E Dinger, Ranjan J Perera
Oncotarget 2014 October 15, 5 (19): 8959-69
25344859
Expression of the long noncoding RNA (lncRNA) SPRY4-IT1 is low in normal human melanocytes but high in melanoma cells. siRNA knockdown of SPRY4-IT1 blocks melanoma cell invasion and proliferation, and increases apoptosis. To investigate its function further, we affinity purified SPRY4-IT1 from melanoma cells and used mass spectrometry to identify the protein lipin 2, an enzyme that converts phosphatidate to diacylglycerol (DAG), as a major binding partner. SPRY4-IT1 knockdown increases the accumulation of lipin2 protein and upregulate the expression of diacylglycerol O-acyltransferase 2 (DGAT2) an enzyme involved in the conversion of DAG to triacylglycerol (TAG). When SPRY4-IT1 knockdown and control melanoma cells were subjected to shotgun lipidomics, an MS-based assay that permits the quantification of changes in the cellular lipid profile, we found that SPRY4-IT1 knockdown induced significant changes in a number of lipid species, including increased acyl carnitine, fatty acyl chains, and triacylglycerol (TAG). Together, these results suggest the possibility that SPRY4-IT1 knockdown may induce apoptosis via lipin 2-mediated alterations in lipid metabolism leading to cellular lipotoxicity.

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