J Pathol Transl Med.  2015 Mar;49(2):105-111. 10.4132/jptm.2015.01.19.

Overexpression of C-reactive Protein as a Poor Prognostic Marker of Resectable Hepatocellular Carcinomas

• Shin JH
• Kim CJ
• Jeon EJ
• Sung CO
• Shin HJ
• Choi J
• Yu E

• KMID: 2381364
• DOI: http://doi.org/10.4132/jptm.2015.01.19

Abstract

BACKGROUND
C-reactive protein (CRP) is an acute phase reactant synthesized in the liver. CRP immunoreactivity is a feature of inflammatory hepatocellular adenomas with a higher risk of malignant transformation. A high serum CRP level denotes poor prognosis in hepatocellular carcinoma (HCC) patients. This study was conducted to determine whether CRP is produced in HCC and to assess the clinicopathologic significance of CRP expression in cancer cells.
METHODS
CRP immunoreactivity was examined in treatment-naive HCCs (n=224) using tissue microarrays and was correlated with clinicopathologic parameters. The expression of CRP mRNA and protein was also assessed in 12 HCC cases by quantitative real-time polymerase chain reaction and immunoblotting. Hep3B and SNU-449 HCC cell lines were used for the analysis of CRP mRNA regulation by interleukin 6 (IL-6).
RESULTS
CRP was expressed in 133 of 224 HCCs (59.4%) with a variable degree of immunoreactivity (grade 1 in 25.9%; grade 2 in 20.1%; grade 3 in 13.4%). There was an inverse relationship between grade 3 CRP immunoreactivity and cancer-specific survival (p=.0047), while no associations were found with other parameters, including recurrence-free survival. The CRP mRNA expression level was significantly higher in CRP immunopositive cases than in immunonegative cases (p<.05). CRP mRNA expression was increased in Hep3B cells, but was not detected in SNU-449 cells even after IL-6 treatment.
CONCLUSIONS
We report the expression of CRP in HCC for the first time. CRP expression was associated with poor cancer-specific survival in patients with resectable HCC.

Keyword

Carcinoma, hepatocellular; C-reactive protein; Immunohistochemistry; Prognosis

MeSH Terms

Adenoma, Liver Cell
C-Reactive Protein*
Carcinoma, Hepatocellular*
Cell Line
Humans
Immunoblotting
Immunohistochemistry
Interleukin-6
Liver
Prognosis
Real-Time Polymerase Chain Reaction
RNA, Messenger
C-Reactive Protein
Interleukin-6
RNA, Messenger

Figure

• 

  Fig. 1. Cytoplasmic C-reactive protein (CRP) immunoreactivity in hepatocellular carcinoma cases. Immunoreactivity is analyzed using a 4-tier grading system: grade 0 (A), grade 1 (B), grade 2 (C), and grade 3 (D).

• 

  Fig. 2. Clinical significance of C-reactive protein (CRP) immunoreactivity. (A) There is a significant difference in cancer-specific survival between patients with CRP grade 3 hepatocellular carcinomas (HCCs) and those with CRP grade 0, 1, and 2 HCCs. (B) There is no difference in recurrence-free survival between the two groups.

• 

  Fig. 3. Correlation between C-reactive protein (CRP) mRNA and protein expression in hepatocellular carcinomas (HCCs). (A) Immunoblotting for CRP protein shows variable expression in HCCs (N, non-neoplastic liver; T, HCC). CRP protein expression is not found in the tumor of case 4, while CRP bands are readily detectable in both non-neoplastic and HCC samples of cases 1, 2, and 3. (B) Quantitative real-time polymerase chain reaction results are shown in box plots of ΔCt for CRP mRNA expression (Ct_CRP–Ct_RPLPO).

• 

  Fig. 4. Induction of C-reactive protein (CRP) mRNA in Hep3B and SNU-449 hepatocellular carcinoma cell lines after interleukin 6 (IL-6) treatment. (A) There are no significant changes in cellular morphology in either cell line after IL-6 treatment. (B) There is a 17.5-fold increase in CRP mRNA expression in IL-6–treated Hep3B cells (50 ng/mL for 6 hours), while CRP mRNA expression is not detected in SNU-449 cells. The yaxis represents fold-changes in CRP mRNA expression following IL-6 treatment.

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