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Identification of a bona fide microRNA biomarker in serum exosomes that predicts hepatocellular carcinoma recurrence after liver transplantation

Sugimachi, K, Matsumura, T, Hirata, H, Uchi, R, Ueda, M, Ueo, H, Shinden, Y, Iguchi, T, Eguchi, H, Shirabe, K, Ochiya, T, Maehara, Y, Mimori, K·2015·British Journal of Cancer
DOI 10.1038/bjc.2014.621PMC PMC4453648
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Figure 1 Exosomal miR in the serum was associated with malignant progression of HCC. (A, B) Exosomes were purified from serum of HCC patients using ultracentrifuge method and examined by electron microscopy; (A) scale bar, 200 nm and (B) scale bar, 100 nm. (C) The exosome extract from serum after an ultracentrifuge method was analysed by western blotting. The data demonstrated that extracts were enriched with exosomal marker protein TSG101. Supernatant of cultured cancer cell lines were used as control. (D–G) miR-718 expression levels in the exosomes from serum of HCC patients who underwent LT were examined according to clinicopathological parameters. (D) Tumour diameter, (E) histological differentiation, (F) vascular invasion, (G) recurrence of the disease. Data represent the means±s.d.
Figure 1 Exosomal miR in the serum was associated with malignant progression of HCC. (A, B) Exosomes were purified from serum of HCC patients using ultracentrifuge method and examined by electron microscopy; (A) scale bar, 200 nm and (B) scale bar, 100 nm. (C) The exosome extract from serum after an ultracentrifuge method was analysed by western blotting. The data demonstrated that extracts were enriched with exosomal marker protein TSG101. Supernatant of cultured cancer cell lines were used as control. (D–G) miR-718 expression levels in the exosomes from serum of HCC patients who underwent LT were examined according to clinicopathological parameters. (D) Tumour diameter, (E) histological differentiation, (F) vascular invasion, (G) recurrence of the disease. Data represent the means±s.d.CC BY-NC-SA · PubMed Central
Figure 2 miR-718 inhibited cell proliferation and targeted HOXB8 gene. (A, B) Induction of miR-718 inhibited the proliferation of HCC cells. (A) miR-718 expression levels in HuH7 cells transfected with pre-miR-718 were significantly higher than those of negative control (nc) cells. (B) The cell proliferation ratio of HuH7 cells transfected with pre-miR-718 were significantly suppressed compared to the negative control (*P<0.01). (C) Overall survival curves of the low and high HOXB8 expression groups for 58 HCC patients who underwent hepatectomy. The patients with high HOXB8 expression showed significantly poorer long-term prognosis after hepatectomy.
Figure 2 miR-718 inhibited cell proliferation and targeted HOXB8 gene. (A, B) Induction of miR-718 inhibited the proliferation of HCC cells. (A) miR-718 expression levels in HuH7 cells transfected with pre-miR-718 were significantly higher than those of negative control (nc) cells. (B) The cell proliferation ratio of HuH7 cells transfected with pre-miR-718 were significantly suppressed compared to the negative control (*P<0.01). (C) Overall survival curves of the low and high HOXB8 expression groups for 58 HCC patients who underwent hepatectomy. The patients with high HOXB8 expression showed significantly poorer long-term prognosis after hepatectomy.CC BY-NC-SA · PubMed Central
Figure 3 miR-718 inhibited the transcription of HOXB8 gene. (A) Sequences of the miR-718 binding sites in the 3′UTRs of transcripts encoding HOXB8. (B) Luciferase analyses in HuH7 and PLC/PRF/5 cell lines. HOXB8 3′ UTR luciferase vector+miR-718 transfectants showed lower luciferase activities than did control cells. Relative luciferase levels were calculated by the following: (Sample Luc/Sample Renilla)/(Control Luc/Control Renilla). Luc, raw firefly luciferase activity; Renilla, Renilla activity (internal transfection control); Pre-miR n.c., pre-miR negative control.
Figure 3 miR-718 inhibited the transcription of HOXB8 gene. (A) Sequences of the miR-718 binding sites in the 3′UTRs of transcripts encoding HOXB8. (B) Luciferase analyses in HuH7 and PLC/PRF/5 cell lines. HOXB8 3′ UTR luciferase vector+miR-718 transfectants showed lower luciferase activities than did control cells. Relative luciferase levels were calculated by the following: (Sample Luc/Sample Renilla)/(Control Luc/Control Renilla). Luc, raw firefly luciferase activity; Renilla, Renilla activity (internal transfection control); Pre-miR n.c., pre-miR negative control.CC BY-NC-SA · PubMed Central

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Metadata

Authors
Sugimachi, K., Matsumura, T., Hirata, H., Uchi, R., Ueda, M., Ueo, H., Shinden, Y., Iguchi, T., Eguchi, H., Shirabe, K., Ochiya, T., Maehara, Y., & Mimori, K.
Venue
British Journal of Cancer
Publisher
Springer Science and Business Media LLC
Published
2015-01-13
Volume
112
Issue
3
Pages
532-538
ISSN
  • 0007-0920
  • 1532-1827
DOI
10.1038/bjc.2014.621
PMC
PMC4453648
Cited by
263
References
2
Language
en
Licence
www.springer.com/tdm (unspecified)

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