Pyruvate kinase M2 is a novel diagnostic marker and predicts tumor progression in human biliary tract cancer

Cancer. 2013 Feb 1;119(3):575-85. doi: 10.1002/cncr.27611. Epub 2012 Aug 1.


Background: The early diagnosis of biliary tract cancer (BTC) remains challenging, and there are few effective therapies. This study investigated whether the M2 isotype of pyruvate kinase (M2-PK), which serves as the key regulator of cellular energy metabolism in proliferating cells, could play a role in the diagnosis and therapy of BTC.

Methods: Plasma and bile M2-PK concentrations were measured by enzyme-linked immunosorbent assay in 88 patients with BTC, 79 with benign biliary diseases, and 17 healthy controls. M2-PK expression was assayed in a BTC tissue array by immunohistochemistry. The role of M2-PK in tumor growth, invasion, and angiogenesis was evaluated in BTC cell lines by retrovirus-mediated M2-PK transfection and short hairpin RNA silencing techniques.

Results: Sensitivity (90.3%) and specificity (84.3%) of bile M2-PK for malignancy were significantly higher than those for plasma M2-PK and serum carbohydrate antigen 19-9. M2-PK expression was specific for cancer cells and correlated with microvessel density. M2-PK positivity was a significant independent prognostic factor by multivariable analysis. Transfection of M2-PK in a negatively expressed cell line (HuCCT-1 cells) increased cell invasion, whereas silencing in an M2-PK-positive cell line (TFK cells) decreased tumor nodule formation and cellular invasion. A significant increase in endothelial tube formation was noted when supernatants from M2-PK-transfected cells were added to an in vitro angiogenesis assay, whereas supernatants from silenced cells negated endothelial tube formation.

Conclusions: Bile M2-PK is a novel tumor marker for BTC and correlates with tumor aggressiveness and poor outcome. Short hairpin RNA-mediated inhibition of M2-PK indicates the potential of M2-PK as a therapeutic target.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Bile / chemistry
  • Bile / metabolism
  • Biliary Tract Neoplasms / diagnosis*
  • Biliary Tract Neoplasms / genetics
  • Biliary Tract Neoplasms / mortality
  • Biliary Tract Neoplasms / pathology
  • Biomarkers, Tumor* / blood
  • Biomarkers, Tumor* / genetics
  • Biomarkers, Tumor* / metabolism
  • Biomarkers, Tumor* / physiology
  • Carcinoma / diagnosis*
  • Carcinoma / genetics
  • Carcinoma / mortality
  • Carcinoma / pathology
  • Case-Control Studies
  • Cell Line, Tumor
  • Disease Progression
  • Female
  • Humans
  • Male
  • Middle Aged
  • Prognosis
  • Pyruvate Kinase / blood
  • Pyruvate Kinase / genetics
  • Pyruvate Kinase / metabolism
  • Pyruvate Kinase / physiology*
  • Survival Analysis


  • Biomarkers, Tumor
  • Pyruvate Kinase