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Viral non-coding RNA inhibits HNF4α expression in HCV associated hepatocellular carcinoma.

Wang Z, Ceniccola K, Florea L, Wang BD, Lee NH, Kumar A - Infect. Agents Cancer (2015)

Bottom Line: Results show inhibition of HNF4α expression by targeting of HNF4α 3'-UTR by HCV-derived small non-coding RNA, vmr11.Vmr11 enhances the invasive properties of HCV-infected cells.These results suggest a direct viral role in the development of hepatocellular carcinoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, The George Washington University, Washington, DC USA.

ABSTRACT

Background: Hepatitis C virus (HCV) infection is an established cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma (HCC); however, it is unclear if the virus plays a direct role in the development of HCC. Hepatocyte nuclear factor 4α (HNF4α) is critical determinant of epithelial architecture and hepatic development; depletion of HNF4α is correlated with oncogenic transformation. We explored the viral role in the inhibition of HNF4α expression, and consequent induction of tumor-promoting genes in HCV infection-associated HCC.

Methods: Western blot analysis was used to monitor the changes in expression levels of oncogenic proteins in liver tissues from HCV-infected humanized mice. The mechanism of HNF4α depletion was studied in HCV-infected human hepatocyte cultures in vitro. Targeting of HNF4α expression by viral non-coding RNA was examined by inhibition of Luciferase HNF4α 3'-UTR reporter. Modulation of invasive properties of HCV-infected cells was examined by Matrigel cell migration assay.

Results: Results show inhibition of HNF4α expression by targeting of HNF4α 3'-UTR by HCV-derived small non-coding RNA, vmr11. Vmr11 enhances the invasive properties of HCV-infected cells. Loss of HNF4α in HCV-infected liver tumors of humanized mice correlates with the induction of epithelial to mesenchymal transition (EMT) genes.

Conclusions: We show depletion of HNF4α in liver tumors of HCV-infected humanized mice by HCV derived small non-coding RNA (vmr11) and resultant induction of EMT genes, which are critical determinants of tumor progression. These results suggest a direct viral role in the development of hepatocellular carcinoma.

No MeSH data available.


Related in: MedlinePlus

Inhibition of HNF4α in human hepatocytes transfected with HCV genomic RNA. Upper part (a) shows Western blots of HNF4α (from left to right), from mock transfected control cells, cells transfected with 1 μg HCV (H77) genomic RNA and (far right lane), cells transfected with 1 μg HCV RNA plus 50nM LNA-vmR-11antagomir. Cells were harvested 48 h post transfection. Lower part (b) shows relative change in HNF4α protein levels as compared to the mock-transfected cells (analyzed in triplicate Mean =/- SE, *p < 0.03)
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Fig3: Inhibition of HNF4α in human hepatocytes transfected with HCV genomic RNA. Upper part (a) shows Western blots of HNF4α (from left to right), from mock transfected control cells, cells transfected with 1 μg HCV (H77) genomic RNA and (far right lane), cells transfected with 1 μg HCV RNA plus 50nM LNA-vmR-11antagomir. Cells were harvested 48 h post transfection. Lower part (b) shows relative change in HNF4α protein levels as compared to the mock-transfected cells (analyzed in triplicate Mean =/- SE, *p < 0.03)

Mentions: To assess a possible direct role of HCV-derived vmr11 in the depletion of HNF4α, we first determined the effects of vmr11 on HNF4α depletion in human hepatocytes transfected with HCV genomic RNA. Results (shown in Fig. 3) indicate that the loss of HNF4α protein in cells transfected with HCV genomic RNA is largely restored by the introduction of antisense vmr11 oligonucleotides. Interestingly, we observed similar decline in HNF4α protein levels of cells transfected with vmr11 oligonucleotides alone. Competition of the effects of vmr11 RNA in vivo by co-transfection of wild-type vmr11 and antisense vmr11 oligonucleotides restored HNF4α protein levels to that of mock-transfected control cells (Fig. 4). The results suggest that the loss of HNF4α protein, either in HCV replicating cells (transfected with HCV genomic RNA), or in cells transduced with vmr11 oligonucleotides alone, is due to the effects of vmr11 RNA. The results suggest that HCV-derived vmr11 RNA contributes to the loss of HNF4α protein in HCV-infected human hepatocytes.Fig. 3


Viral non-coding RNA inhibits HNF4α expression in HCV associated hepatocellular carcinoma.

Wang Z, Ceniccola K, Florea L, Wang BD, Lee NH, Kumar A - Infect. Agents Cancer (2015)

Inhibition of HNF4α in human hepatocytes transfected with HCV genomic RNA. Upper part (a) shows Western blots of HNF4α (from left to right), from mock transfected control cells, cells transfected with 1 μg HCV (H77) genomic RNA and (far right lane), cells transfected with 1 μg HCV RNA plus 50nM LNA-vmR-11antagomir. Cells were harvested 48 h post transfection. Lower part (b) shows relative change in HNF4α protein levels as compared to the mock-transfected cells (analyzed in triplicate Mean =/- SE, *p < 0.03)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4495692&req=5

Fig3: Inhibition of HNF4α in human hepatocytes transfected with HCV genomic RNA. Upper part (a) shows Western blots of HNF4α (from left to right), from mock transfected control cells, cells transfected with 1 μg HCV (H77) genomic RNA and (far right lane), cells transfected with 1 μg HCV RNA plus 50nM LNA-vmR-11antagomir. Cells were harvested 48 h post transfection. Lower part (b) shows relative change in HNF4α protein levels as compared to the mock-transfected cells (analyzed in triplicate Mean =/- SE, *p < 0.03)
Mentions: To assess a possible direct role of HCV-derived vmr11 in the depletion of HNF4α, we first determined the effects of vmr11 on HNF4α depletion in human hepatocytes transfected with HCV genomic RNA. Results (shown in Fig. 3) indicate that the loss of HNF4α protein in cells transfected with HCV genomic RNA is largely restored by the introduction of antisense vmr11 oligonucleotides. Interestingly, we observed similar decline in HNF4α protein levels of cells transfected with vmr11 oligonucleotides alone. Competition of the effects of vmr11 RNA in vivo by co-transfection of wild-type vmr11 and antisense vmr11 oligonucleotides restored HNF4α protein levels to that of mock-transfected control cells (Fig. 4). The results suggest that the loss of HNF4α protein, either in HCV replicating cells (transfected with HCV genomic RNA), or in cells transduced with vmr11 oligonucleotides alone, is due to the effects of vmr11 RNA. The results suggest that HCV-derived vmr11 RNA contributes to the loss of HNF4α protein in HCV-infected human hepatocytes.Fig. 3

Bottom Line: Results show inhibition of HNF4α expression by targeting of HNF4α 3'-UTR by HCV-derived small non-coding RNA, vmr11.Vmr11 enhances the invasive properties of HCV-infected cells.These results suggest a direct viral role in the development of hepatocellular carcinoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, The George Washington University, Washington, DC USA.

ABSTRACT

Background: Hepatitis C virus (HCV) infection is an established cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma (HCC); however, it is unclear if the virus plays a direct role in the development of HCC. Hepatocyte nuclear factor 4α (HNF4α) is critical determinant of epithelial architecture and hepatic development; depletion of HNF4α is correlated with oncogenic transformation. We explored the viral role in the inhibition of HNF4α expression, and consequent induction of tumor-promoting genes in HCV infection-associated HCC.

Methods: Western blot analysis was used to monitor the changes in expression levels of oncogenic proteins in liver tissues from HCV-infected humanized mice. The mechanism of HNF4α depletion was studied in HCV-infected human hepatocyte cultures in vitro. Targeting of HNF4α expression by viral non-coding RNA was examined by inhibition of Luciferase HNF4α 3'-UTR reporter. Modulation of invasive properties of HCV-infected cells was examined by Matrigel cell migration assay.

Results: Results show inhibition of HNF4α expression by targeting of HNF4α 3'-UTR by HCV-derived small non-coding RNA, vmr11. Vmr11 enhances the invasive properties of HCV-infected cells. Loss of HNF4α in HCV-infected liver tumors of humanized mice correlates with the induction of epithelial to mesenchymal transition (EMT) genes.

Conclusions: We show depletion of HNF4α in liver tumors of HCV-infected humanized mice by HCV derived small non-coding RNA (vmr11) and resultant induction of EMT genes, which are critical determinants of tumor progression. These results suggest a direct viral role in the development of hepatocellular carcinoma.

No MeSH data available.


Related in: MedlinePlus