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, 9 (12), 2105-2116

IL-8 Induces Transdifferentiation of Mature Hepatocytes Toward the Cholangiocyte Phenotype

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IL-8 Induces Transdifferentiation of Mature Hepatocytes Toward the Cholangiocyte Phenotype

Tokio Sasaki et al. FEBS Open Bio.

Abstract

The adult mammalian liver exhibits a remarkable regenerative capacity, with different modes of regeneration according to the type and extent of injury. Hepatocyte-cholangiocyte biphenotypic liver progenitor cell populations appear under conditions of excessive injury. It has been reported that mature hepatocytes can transdifferentiate toward a cholangiocyte phenotype and be a cellular source of progenitor cell populations. Here, we determined that among various plasma cytokines, interleukin (IL)-8 levels were significantly elevated in acute liver failure and severe acute liver injury patients. In vitro assays revealed that administration of IL-8 homologues increases the expression of Sry HMG box protein 9 (SOX9). In liver biopsies of acute liver injury patients, we observed the appearance of SOX9-positive biphenotypic hepatocytes accompanied by elevation of plasma IL-8 levels. Our results suggest that IL-8 regulates the phenotypic conversion of mature hepatocytes toward a cholangiocyte phenotype.

Keywords: Sry HMG box protein 9; cholangiocyte; interleukin-8; mature hepatocyte; transdifferentiation.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Serum cytokine levels of ALF and SLI patients. Results are presented as median and interquartile range.
Figure 2
Figure 2
Effects of KC and MIP‐2 on the proliferative activity of mouse liver cell lines. (A) Cxcr1 and Cxcr2 mRNA levels in AML12, LPCs, and 603B cells. Glyceraldehyde 3‐phosphate dehydrogenase (Gapdh) mRNA was used as the internal control. (B) Cell viability of AML12, LPCs, and 603B cells were estimated in response to administration with KC or MIP‐2 (100 pg·mL−1, 1 ng·mL−1, or 10 ng·mL−1). Student's unpaired t‐test was performed to compare treated and control samples. Results are presented as mean ± SEM (n = 3), *P < 0.05. (C) Western blotting analysis was performed using cell lysates of AML12 and LPCs and antibodies specific for cyclin D1 as indicated. Both AML12 and LPCs were treated with KC or MIP‐2 (100 pg·mL−1, 1 ng·mL−1, or 10 ng·mL−1). β‐Actin was used as the internal control.
Figure 3
Figure 3
Gene expression and immunohistochemical analysis of SOX9. (A) Timeline of experimental protocols of AML12 for quantitative reverse transcription PCR (qRT‐PCR) and immunofluorescence labeling of cells. AML12 cells were seeded at 5.0 × 104 cells·mL−1 into 6‐well plates (1.5 × 105 cells/well). After incubation for 24 h, medium with 10 ng·mL−1 of KC or MIP‐2 was changed at 24 and 72 h. (B) Sox9 mRNA levels in AML12 cells collected after 120 h of incubation was determined using qRT‐PCR. mRNA levels were normalized using 18s rRNA as a housekeeping gene. Student's unpaired two‐tailed t‐test was performed. Results are represented as mean ± SEM (n = 3). (C) Immunofluorescent staining of SOX9 in AML12 cells treated with or without KC and MIP‐2 after 120 h of incubation. The number of SOX9‐positive cells was quantified per 50 cells. Fisher's exact test was performed to compare treated and control samples. Scale bar, 100 μm. (D) Timeline of the experimental protocols of primary hepatocytes for qRT‐PCR and immunofluorescence labeling of cells. Primary hepatocytes were seeded at 1.0 × 105 cells·mL−1 into 6‐well plates (2.0 × 105 cells/well). After incubation for 24 h, medium with 10 ng·mL−1 of KC or MIP‐2 was changed at 24 and 48 h. (E) Sox9 mRNA levels in primary hepatocytes collected after 72 h of incubation was determined using qRT‐PCR. mRNA levels were normalized using 18s rRNA as a housekeeping gene. Student's unpaired two‐tailed t‐test was performed. Results are represented as mean ± SEM (n = 3). (F) Immunofluorescent staining of SOX9 in primary hepatocytes treated with or without KC and MIP‐2 after 72 h of incubation. Primary hepatocytes were cultured in the absence or presence of 10 ng·mL−1 of KC or MIP‐2. The number of SOX9‐positive cells was quantified per 50 cells. Fisher's exact test was performed to compare treated and control samples. Scale bar, 100 μm. PH, primary hepatocyte.
Figure 4
Figure 4
Colorimetric immunohistochemical staining and immunofluorescent detection of liver biopsy specimens. (A) SOX9 staining of liver biopsy specimens of ALI and SS patients. Mean ± SEM (SS, n = 3; ALI, n = 8). (B) A 53‐year‐old woman with autoimmune hepatitis. (C) A 35‐year‐old man with acute hepatitis A. (D) A 64‐year‐old man with acute hepatitis B. Scale bars: 100 μm for colorimetric immunohistochemical staining images and 30 µm for immunofluorescent staining images. SS, simple steatosis.

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References

    1. Edwards JL and Klein RE (1961) Cell renewal in adult mouse tissues. Am J Pathol 38, 437–453. - PMC - PubMed
    1. Wabik A and Jones PH (2015) Switching roles: the functional plasticity of adult tissue stem cells. Embo J 34, 1164–1179. - PMC - PubMed
    1. Wang B, Zhao L, Fish M, Logan CY and Nusse R (2015) Self‐renewing diploid Axin2(+) cells fuel homeostatic renewal of the liver. Nature 524, 180–185. - PMC - PubMed
    1. Malato Y, Naqvi S, Schurmann N, Ng R, Wang B, Zape J, Kay MA, Grimm D and Willenbring H (2011) Fate tracing of mature hepatocytes in mouse liver homeostasis and regeneration. J Clin Invest 121, 4850–4860. - PMC - PubMed
    1. Miyaoka Y, Ebato K, Kato H, Arakawa S, Shimizu S and Miyajima A (2012) Hypertrophy and unconventional cell division of hepatocytes underlie liver regeneration. Curr Biol 22, 1166–1175. - PubMed

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