The lncRNA H19-Derived MicroRNA-675 Promotes Liver Necroptosis by Targeting FADD

SIMPLE SUMMARY: Liver cancer develops mostly in a chronically inflamed liver. The inflammation process can enhance or suppress the development of liver cancer. H19 is a noncoding RNA that is upregulated in inflamed livers. The role of H19 in liver cancer was intensely investigated, but the reported findings are conflicting. Some reported that H19 is a tumor suppressor and others that it has oncogenic properties. To understand the contribution of H19 to liver cancer development, we investigated miR-675, which is generated from the first exon of the H19 RNA message. Interestingly, we found that miR-675 suppresses liver cancer cell growth by inducing cell death. Following our investigation of the mechanism of this killing effect, we established that miR-675 represses the protein called Fas-associated protein with death domain (FADD) and that this repression leads to the development of a specific type of necrosis named necroptosis. These findings can have future therapeutic implications. ABSTRACT: The H19-derived microRNA-675 (miR-675) has been implicated as both tumor promoter and tumor suppressor and also plays a role in liver inflammation. We found that miR-675 promotes cell death in human hepatocellular carcinoma (HCC) cell lines. We show that Fas-associated protein with death domain (FADD), a mediator of apoptotic cell death signaling, is downregulated by miR-675 and a negative correlation exists between miR-675 and FADD expression in mouse models of HCC (p = 0.014) as well as in human samples (p = 0.017). We demonstrate in a mouse model of liver inflammation that overexpression of miR-675 promotes necroptosis, which can be inhibited by the necroptosis-specific inhibitor Nec-1/Nec-1s. miR-675 induces the level of both p-MLKL (Mixed Lineage Kinase Domain-Like Pseudokinase) and RIP3 (receptor-interacting protein 3), which are key signaling molecules in necroptosis, and enhances MLKL binding to RIP3. miR-675 also inhibits the levels of cleaved caspases 8 and 3, suggesting that miR-675 induces a shift from apoptosis to a necroptotic cellular pathway. In conclusion, downregulation of FADD by miR-675 promotes liver necroptosis in response to inflammatory signals. We propose that this regulation cascade can stimulate and enhance the inflammatory response in the liver, making miR-675 an important regulator in liver inflammation and potentially also in HCC.