Korean J Physiol Pharmacol.  2013 Oct;17(5):455-461. 10.4196/kjpp.2013.17.5.455.

The Effect of Metformin Treatment on CRBP-I Level and Cancer Development in the Liver of HBx Transgenic Mice

• Kim JH
• Alam MM
• Park DB
• Cho M
• Lee SH
• Jeon YJ
• Yu DY
• Kim TD
• Kim HY
• Cho CG
• Lee DH

• KMID: 1500242
• DOI: http://doi.org/10.4196/kjpp.2013.17.5.455

Abstract

Retinoids regulate not only various cell functions including proliferation and differentiation but also glucose and lipid metabolism. After we observed a marked up-regulation of cellular retinol-binding protein-I (CRBP-I) in the liver of hepatitis B virus x antigen (HBx)-transgenic (HBx Tg) mice which are prone to hepatocellular carcinoma (HCC) and fatty liver, we aimed to evaluate retinoid pathway, including genes for the retinoid physiology, CRBP-I protein expression, and retinoid levels, in the liver of HBx Tg mice. We also assessed the effect of chronic metformin treatment on HCC development in the mice. Many genes involved in hepatic retinoid physiology, including CRBP-I, were altered and the tissue levels of retinol and all-trans retinoic acid (ATRA) were elevated in the liver of HBx Tg mice compared to those of wild type (WT) control mice. CRBP-I protein expression in liver, but not in white adipose tissue, of HBx Tg mice was significantly elevated compared to WT control mice while CRBP-I protein expressions in the liver and WAT of high-fat fed obese and db/db mice were comparable to WT control mice. Chronic treatment of HBx Tg mice with metformin did not affect the incidence of HCC, but slightly increased hepatic CRBP-I level. In conclusion, hepatic CRBP-I level was markedly up-regulated in HCC-prone HBx Tg mice and neither hepatic CRBP-I nor the development of HCC was suppressed by metformin treatment.

Keyword

Cellular retinol-binding protein-I; HBx protein; Hepatocellular carcinoma; Metformin; Retinoids

MeSH Terms

Adipose Tissue, White
Animals
Carcinoma, Hepatocellular*
Fatty Liver
Hepatitis B virus
Incidence
Lipid Metabolism
Liver*
Metformin*
Mice
Mice, Transgenic*
Retinoids
Retinol-Binding Proteins, Cellular*
Trans-Activators
Tretinoin
Up-Regulation
Vitamin A
Glucose
Metformin
Retinoids
Trans-Activators
Tretinoin
Vitamin A

Figure

• 

  Fig. 1 Real-time PCR analysis of the genes involved in retinoid metabolism (A) and retinoid signaling and other physiologic pathway (B) in the liver of WT and HBx Tg mice. The expression level of each gene was normalized to GAPDH. *p<0.05; **p<0.01 vs. WT mice.

• 

  Fig. 2 CRBP-I protein expression in liver and eWAT of WT control mice (C), HF-fed control mice (C-HF), db/db mice, and HBx Tg mice (n=6 mice in each group). (A) The representative immunoblotting images. (B, C) The levels of protein expression presented as the percentage of the mean expression level of the WT control group (% of control). Alphabetical letters on the bars indicated statistical differences (p<0.05) among the experimental groups: the same letter indicates no statistical difference.

• 

  Fig. 3 Hepatic retinol (A) and ATRA (B) levels in WT control and HBx Tg mice. The level of retinoid was presented as µg per gram of dry extract. *p<0.05 vs. WT control mice.

• 

  Fig. 4 Hepatic expression levels of Akt, CRBP-I, and pAMPK in WT and HBx Tg mice, and metformin-treated HBx Tg mice (Met-HBx Tg). The representative immunoblotting images (A) and the levels of protein expression were presented (% of control) (B). *p<0.05; **p<0.01 vs. WT mice. †p<0.05; ††p<0.01 vs. HBx Tg mice.

• 

  Fig. 5 The fatty liver change and the development of HCC in HBx Tg mice. Hematoxylin and eosin-stained representative images are presented. (A) Relatively normal-looking portion in the liver of HBx Tg mice (×100). (B) Fatty changes and chronic inflammatory cell infiltrations are noted in the dysplastic liver of HBx Tg mice (×100). (C) Microscopic appearance of grossly identified HCC. The un-encapsulated and well differentiated HCC lesion has a trabecular pattern and is compressing the surrounding dysplastic hepatocytes (×40). Arrows indicate the boundary of HCC. (D) Microscopic appearance of grossly identified HCC shows nuclear pleomorphism, nucleomegaly and frequent mitoses (×200). Arrows indicate mitosis.

• 

  Fig. 6 The summary of retinoid pathway. TTR, transthyretin.

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