Loss of p16 contributes to p27 sequestration by cyclin D(1)-cyclin-dependent kinase 4 complexes and poor prognosis in hepatocellular carcinoma

Clin Cancer Res. 2003 Aug 15;9(9):3389-96.

Abstract

Purpose: p27(Kip1) (p27) might act as an adverse prognostic marker for various types of cancers. However, its clinical usefulness remains uncertain, because it is sometimes overexpressed in aggressive types.

Experimental design: To precisely evaluate the practical significance of p27 in hepatocellular carcinoma (HCC), we immunohistochemically compared the level of p27 expression with Ki-67 labeling in 74 HCCs and focused on tumors in which cell proliferation increased despite a high level of p27 expression. We then analyzed the status of p27 and related cell-cycle regulators using kinase and immunoprecipitation assays, Western blotting, and methylation-specific PCR to understand the rationale for the functional inactivation of p27 in HCC. We also evaluated relationships between the key biological characteristics of HCC and survival.

Results: Immunohistochemical studies showed that 40 (54%) of 74 HCCs expressed high levels of p27 (>50% of the tumor cells). Of these, the Ki-67 labeling index was low (<20%) in 26 (65%) and high (>20%) in 14 (35%). Increased proliferative activities were closely correlated with elevated kinase activities, sequestration of p27 protein, and p16 gene methylation. The association between a loss of p16 and poor prognosis was significant when p27 expression was high (P < 0.01).

Conclusions: The loss of p16 appears to be closely related to the functional inactivation of p27, and assessment of p16 status may be useful for a precise prognostic prediction of individuals with HCC expressing high levels of p27.

MeSH terms

  • Adult
  • Aged
  • Blotting, Western
  • Carcinoma, Hepatocellular / metabolism*
  • Cell Cycle
  • Cell Cycle Proteins / metabolism*
  • Cell Division
  • Cell Line, Tumor
  • Cyclin D1 / metabolism*
  • Cyclin-Dependent Kinase 4
  • Cyclin-Dependent Kinase Inhibitor p16 / genetics*
  • Cyclin-Dependent Kinase Inhibitor p27
  • Cyclin-Dependent Kinases / metabolism*
  • DNA Methylation
  • Female
  • Humans
  • Immunohistochemistry
  • Ki-67 Antigen / biosynthesis
  • Liver Neoplasms / metabolism*
  • Male
  • Middle Aged
  • Polymerase Chain Reaction
  • Precipitin Tests
  • Prognosis
  • Promoter Regions, Genetic
  • Proto-Oncogene Proteins*
  • Time Factors
  • Tumor Suppressor Proteins / metabolism*

Substances

  • Cell Cycle Proteins
  • Cyclin-Dependent Kinase Inhibitor p16
  • Ki-67 Antigen
  • Proto-Oncogene Proteins
  • Tumor Suppressor Proteins
  • Cyclin D1
  • Cyclin-Dependent Kinase Inhibitor p27
  • CDK4 protein, human
  • Cyclin-Dependent Kinase 4
  • Cyclin-Dependent Kinases