Association between MicroRNA-373 and Long Noncoding RNA NORAD in Hepatitis C Virus-Infected Hepatocytes Impairs Wee1 Expression for Growth Promotion
Authors: Reina Sasaki
Ratna B Ray
Robert Steele
Subhayan Sur
Pradip Devhare
Ranjit Ray
Journal: Journal of Virology (2018)
Discipline: MICROBIOLOGY
Keywords: NORAD; Wee1; hepatitis C virus; miRNA-373.
Affiliations: Department of Pathology, Saint Louis University, St. Louis, Missouri, USA., Department of Internal Medicine, Saint Louis University, St. Louis, Missouri, USA., Department of Pathology, Saint Louis University, St. Louis, Missouri, USA .

Abstract:  Chronic hepatitis C virus (HCV) infection may lead to end-stage liver disease, including hepatocellular carcinoma (HCC). We have shown previously that microRNA-373 (miR-373) is upregulated in HCV-infected human liver biopsy specimens. To gain insight into the role of miR-373 in HCV-mediated pathogenesis, we investigated its interacting partner for hepatocyte growth regulation. Transcriptome sequencing (RNA-seq) data revealed that Wee1 is associated with miR-373 and is a direct target. Interestingly, higher expression of Wee1 was noted in HCV-infected hepatocytes than in uninfected hepatocytes, suggesting that other factors may block miR-373-mediated Wee1 inhibition. We subsequently found an association between the long noncoding RNA NORAD (LINC00657) and miR-373, and we demonstrated that NORAD binds to miR-373 and Wee1 independently. However, the high level of Wee1 expression in HCV-infected hepatocytes suggested that miR-373 forms a complex with NORAD. Depletion of miR-373 or the inhibitor Wee1 reduces the growth of Huh7.5 cells harboring the HCV genome as well as reducing Wee1 expression. Taken together, our data demonstrate a novel mechanism of hepatocyte growth promotion during HCV infection involving a miR-373-NORAD-Wee1 axis, which may be a target for future therapy against HCV-associated HCC.IMPORTANCE The mechanism of HCV-mediated liver pathogenesis is poorly understood. In this study, we observed that HCV infection upregulates miR-373 and Wee1, a pivotal player in the G2 checkpoint in the cell cycle, although Wee1 is a direct target for miR-373. Subsequent investigation demonstrated that miR-373 forms a complex with the long noncoding RNA NORAD, resulting in the release of their common target, Wee1, in HCV-infected cells, which, in turn, favors uncontrolled cell growth. Our study suggested a previously unknown mechanism for hepatocyte growth promotion following HCV infection, and this pathway can be targeted for future therapy against HCV-mediated liver pathogenesis. 

Retracted: No

PubPeer comments:
 Blue boxes: Control lanes in Figures 1D, 2B, and 4B are unexpectedly similar. Red and orange arrows indicate blemishes that are visible on all three blots. The methods describe having a different control treatment for each of the three groups, and the similarities are even more surprising upon reading that Huh7.5 cells were used for 1D and 2B, IHH were used for 4B ![file] Pink boxes: Control lanes in Figure 5A, 5B, and 5D appear remarkably similar as well. The authors acknowledge in the caption that panels A and B share a control lane; according to the methods section the control should have been different for each experiment (control pcDNA3 for NORAD transfections, control miR for miR-373 transfections, mock infection for HCV) ![file] ,