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Hepatitis B virus-specific miRNAs and Argonaute2 play a role in the viral life cycle.

Hayes CN, Akamatsu S, Tsuge M, Miki D, Akiyama R, Abe H, Ochi H, Hiraga N, Imamura M, Takahashi S, Aikata H, Kawaoka T, Kawakami Y, Ohishi W, Chayama K - PLoS ONE (2012)

Bottom Line: AGO2 and HBcAg were found to physically interact and co-localize in the ER and other subcellular compartments.HBs was also found to co-localize with AGO2 and was detected in multiple subcellular compartments.These results suggest that AGO2 and HBV-specific miRNAs might play a role in the HBV life cycle.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Metabolism, Applied Life Sciences, Institute of Biomedical & Health Sciences, Hiroshima University, Hiroshima, Japan.

ABSTRACT

Unlabelled: Disease-specific serum miRNA profiles may serve as biomarkers and might reveal potential new avenues for therapy. An HBV-specific serum miRNA profile associated with HBV surface antigen (HBsAg) particles has recently been reported, and AGO2 and miRNAs have been shown to be stably associated with HBsAg in serum. We identified HBV-associated serum miRNAs using the Toray 3D array system in 10 healthy controls and 10 patients with chronic hepatitis B virus (HBV) infection. 19 selected miRNAs were then measured by quantitative RT-PCR in 248 chronic HBV patients and 22 healthy controls. MiRNA expression in serum versus liver tissue was also compared using biopsy samples. To examine the role of AGO2 during the HBV life cycle, we analyzed intracellular co-localization of AGO2 and HBV core (HBcAg) and surface (HBsAg) antigens using immunocytochemistry and proximity ligation assays in stably transfected HepG2 cells. The effect of AGO2 ablation on viral replication was assessed using siRNA. Several miRNAs, including miR-122, miR-22, and miR-99a, were up-regulated at least 1.5 fold (P<2E-08) in serum of HBV-infected patients. AGO2 and HBcAg were found to physically interact and co-localize in the ER and other subcellular compartments. HBs was also found to co-localize with AGO2 and was detected in multiple subcellular compartments. Conversely, HBx localized non-specifically in the nucleus and cytoplasm, and no interaction between AGO2 and HBx was detected. SiRNA ablation of AGO2 suppressed production of HBV DNA and HBs antigen in the supernatant.

Conclusion: These results suggest that AGO2 and HBV-specific miRNAs might play a role in the HBV life cycle.

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Related in: MedlinePlus

siRNA knock down of AGO2 expression.A) Knock down of AGO2 expression in T23 cells by specific siRNAs for AGO2 or control siRNAs, confirmed by real-time quantitative RT-PCR analysis. B) Supernatant HBs antigen, and C) HBV-DNA were measured. Both were higher in supernatant of cells transfected with si-control than in cells transfected with si-AGO2. D) There was no significant difference in cell viability between cells transfected with si-control compared to those with si-AGO2.
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pone-0047490-g004: siRNA knock down of AGO2 expression.A) Knock down of AGO2 expression in T23 cells by specific siRNAs for AGO2 or control siRNAs, confirmed by real-time quantitative RT-PCR analysis. B) Supernatant HBs antigen, and C) HBV-DNA were measured. Both were higher in supernatant of cells transfected with si-control than in cells transfected with si-AGO2. D) There was no significant difference in cell viability between cells transfected with si-control compared to those with si-AGO2.

Mentions: Antisense RNA directed against AGO2 strongly suppressed AGO2 expression (Fig. 4A) and resulted in lower HBV DNA (Fig. 4B) and HBsAg (Fig. 4C) levels in the supernatant. Cell viability was not significantly reduced (Fig. 4D).


Hepatitis B virus-specific miRNAs and Argonaute2 play a role in the viral life cycle.

Hayes CN, Akamatsu S, Tsuge M, Miki D, Akiyama R, Abe H, Ochi H, Hiraga N, Imamura M, Takahashi S, Aikata H, Kawaoka T, Kawakami Y, Ohishi W, Chayama K - PLoS ONE (2012)

siRNA knock down of AGO2 expression.A) Knock down of AGO2 expression in T23 cells by specific siRNAs for AGO2 or control siRNAs, confirmed by real-time quantitative RT-PCR analysis. B) Supernatant HBs antigen, and C) HBV-DNA were measured. Both were higher in supernatant of cells transfected with si-control than in cells transfected with si-AGO2. D) There was no significant difference in cell viability between cells transfected with si-control compared to those with si-AGO2.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3472984&req=5

pone-0047490-g004: siRNA knock down of AGO2 expression.A) Knock down of AGO2 expression in T23 cells by specific siRNAs for AGO2 or control siRNAs, confirmed by real-time quantitative RT-PCR analysis. B) Supernatant HBs antigen, and C) HBV-DNA were measured. Both were higher in supernatant of cells transfected with si-control than in cells transfected with si-AGO2. D) There was no significant difference in cell viability between cells transfected with si-control compared to those with si-AGO2.
Mentions: Antisense RNA directed against AGO2 strongly suppressed AGO2 expression (Fig. 4A) and resulted in lower HBV DNA (Fig. 4B) and HBsAg (Fig. 4C) levels in the supernatant. Cell viability was not significantly reduced (Fig. 4D).

Bottom Line: AGO2 and HBcAg were found to physically interact and co-localize in the ER and other subcellular compartments.HBs was also found to co-localize with AGO2 and was detected in multiple subcellular compartments.These results suggest that AGO2 and HBV-specific miRNAs might play a role in the HBV life cycle.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Metabolism, Applied Life Sciences, Institute of Biomedical & Health Sciences, Hiroshima University, Hiroshima, Japan.

ABSTRACT

Unlabelled: Disease-specific serum miRNA profiles may serve as biomarkers and might reveal potential new avenues for therapy. An HBV-specific serum miRNA profile associated with HBV surface antigen (HBsAg) particles has recently been reported, and AGO2 and miRNAs have been shown to be stably associated with HBsAg in serum. We identified HBV-associated serum miRNAs using the Toray 3D array system in 10 healthy controls and 10 patients with chronic hepatitis B virus (HBV) infection. 19 selected miRNAs were then measured by quantitative RT-PCR in 248 chronic HBV patients and 22 healthy controls. MiRNA expression in serum versus liver tissue was also compared using biopsy samples. To examine the role of AGO2 during the HBV life cycle, we analyzed intracellular co-localization of AGO2 and HBV core (HBcAg) and surface (HBsAg) antigens using immunocytochemistry and proximity ligation assays in stably transfected HepG2 cells. The effect of AGO2 ablation on viral replication was assessed using siRNA. Several miRNAs, including miR-122, miR-22, and miR-99a, were up-regulated at least 1.5 fold (P<2E-08) in serum of HBV-infected patients. AGO2 and HBcAg were found to physically interact and co-localize in the ER and other subcellular compartments. HBs was also found to co-localize with AGO2 and was detected in multiple subcellular compartments. Conversely, HBx localized non-specifically in the nucleus and cytoplasm, and no interaction between AGO2 and HBx was detected. SiRNA ablation of AGO2 suppressed production of HBV DNA and HBs antigen in the supernatant.

Conclusion: These results suggest that AGO2 and HBV-specific miRNAs might play a role in the HBV life cycle.

Show MeSH
Related in: MedlinePlus