Eleni Birli
Nottingham Trent University, UK
Title: EMBH1 lncRNA is a major regulator of liver cancer
Biography
Biography: Eleni Birli
Abstract
Statement of the problem: Hepatocellular carcinoma (HCC) is the fourth leading cause of deaths worldwide, indicating the necessity of novel therapeutic approaches. Deregulation of long non-coding RNAs (lncRNAs) has been associated with oncogenic transformation.
Methodology: To identify transcripts involved in HCC chemoresistance, we have established liver cancer cell lines resistant to chemotherapy and performed whole-transcriptome analysis. To investigate the eff ect of lncRNA on cellular properties, lncRNA expression was manipulated and performed a series of functional studies. LncRNA depletion was achieved transiently or stably by means of antisense GapmeRs or shRNAs, respectively. LncRNA stable overexpression was employed through lentiviral transduction. Cell growth was interrogated through luminescence viability, live cell analysis and colony formation assays. Migration and invasion were studied through live cell analysis scratch wound assay and matrigel invasion chambers. Metabolic rate of lncRNA manipulated cells was profi led by real-time bioenergetic assays. To characterize the lncRNA interactome, mass spectrometry was employed following RNA immunoprecipitation. To study the relevance of lncRNA to human disease, expression and cellular localization was assessed by qRT-PCR and in situ hybridization.
Findings: Transcriptome analysis revealed 15,857 up-regulated and 22,332 down-regulated lncRNAs in chemoresistant liver cancer cell lines. Knockdown of the up-regulated lncRNAs followed by functional assays, suggest 31 novel lncRNAs as regulators of HCC. We focused our studies on the lncRNA with the strongest eff ect, EMBH1. EMBH1 silencing signifi cantly inhibits, while EMBH1 overexpression induces, liver cancer cell growth and aggressiveness. In agreement with its oncogenic role, EMBH1 induces a glycolytic metabolic shift a phenomenon known as the Warburg eff ect. Mass spectrometry data identified specific EMBH1 protein interactors. In patients, EMBH1 was identifi ed as a nuclear-dominant lncRNA and was overexpressed in 65 liver cancer human tissues by 100-fold (when compared to 17 normal tissues).
Conclusion & Significance: EMBH1 is a potential novel therapeutic target for HCC progression and aggressiveness.