Favorable survival outcomes in this study. CK catalyzes the production of phosphocholine, a substrate used in the Kennedy (citidine diphosphocholine-choline) pathway to synthesize phosphatidylcholine 
and other membrane phospholipids [36]. Increased CK expression, along with increased intracellular levels of phosphocholine, occurs in a variety of cancers and is ostensibly related to the rate of cellular proliferation in tumors [8,37?1]. Thus, the association between tumor CK expression and survival in this study may be explained 
on the basis of tumor growth. However, several recent studies have not substantiated a direct relationship between tumor proliferative activity and choline metabolism [10,42?4]. An alternative explanation for why increased CK activity can lead to more aggressive tumors is the potential role of phosphocholine as a second-messenger in mitogenic signaling. Specifically, the production of phosphocholine appears necessary for the activation of Raf-1 in the mitogenactivated protein kinase (MAPK) pathway [45], as well as the activation of protein kinase B (PKB/Akt) in the phosphatidylinositol 3-kinase (PI3K)-Akt pathway [46]. This role of phosphocholine in mitogenic signal transduction may therefore also serve as another potential explanation for the association between tumor CKA expression and increased mortality found in the current study. Tumor HK2 expression was a weaker predictor of survival than tumor size and cancer stage in multivariable analysis. The impact of tumor CKA expression was also mitigated after adjustments for other clinicopathologic variables. Therefore, these immunohistochemical markers may have limited clinical value for patient risk stratification beyond the clinical parameters already used to assess patient prognosis. Nonetheless, the current study provides evidence that glycolysis and choline metabolism are biologically relevant to clinical outcomes in HCC, which should encourage further investigation of these metabolic pathways as potential therapeutic targets in HCC. In this study, tumor HK2 expression status identified a subset of patients with less favorable prognosis among patients with CKA-positive tumors. In turn, tumor CKA expression identified patients with worse prognosis among patients with HK2-positive tumors. The biologic basis of these observations may be uncertain since the relationship between glucose metabolism and choline metabolism in cancer is not yet well understood. In breast cancer cells, CKA down-regulation has been shown to exert little effect on cellular FDG metabolism, suggesting that glucose metabolism may not be coupled to choline metabolism [47]. This raises the KDM5A-IN-1 biological activity possibility that glucose metabolism and choline metabolism are independently advantageous to tumor cells. A recent PET imaging trial comparing FDG PET with fluorine-18 labeled choline (FC) PET for the detection of HCC reported that welldifferentiated tumors are more likely to be detected by only FC PET, while less differentiated tumors may be detected by either PET technique with similar efficacy [13]. In view of the current study results, it may be hypothesized that livers tumors showing abnormalities on both FDG and FC PET are potentially more aggressive and lethal. One limitation of this study is that the use of microscopy arrays limits the assessment of HK2 and CKA expression to very small portions of tumor. It was not possible to confirm the clinical tumor grade of these samples given the amount of ti.Favorable survival outcomes in this study. CK catalyzes the production of phosphocholine, a substrate used in the Kennedy (citidine diphosphocholine-choline) pathway to synthesize phosphatidylcholine and other membrane phospholipids [36]. Increased CK expression, along with increased intracellular levels of phosphocholine, occurs in a variety of cancers and is ostensibly related to the rate of cellular proliferation in tumors [8,37?1]. Thus, the association between tumor CK expression and survival in this study may be explained on the basis of tumor growth. However, several recent studies have not substantiated a direct relationship between tumor proliferative activity and choline metabolism [10,42?4]. An alternative explanation for why increased CK activity can lead to more aggressive tumors is the potential role of phosphocholine as a second-messenger in mitogenic signaling. Specifically, the production of phosphocholine appears necessary for the activation of Raf-1 in the mitogenactivated protein kinase (MAPK) pathway [45], as well as the activation of protein kinase B (PKB/Akt) in the phosphatidylinositol 3-kinase (PI3K)-Akt pathway [46]. This role of phosphocholine in mitogenic signal transduction may therefore also serve as another potential explanation for the association between tumor CKA expression and increased mortality found in the current study. Tumor HK2 expression was a weaker predictor of survival than tumor size and cancer stage in multivariable analysis. The impact of tumor CKA expression was also mitigated after adjustments for other clinicopathologic variables. Therefore, these immunohistochemical markers may have limited clinical value for patient risk stratification beyond the clinical parameters already used to assess patient prognosis. Nonetheless, the current study provides evidence that glycolysis and choline metabolism are biologically relevant to clinical outcomes in HCC, which should encourage further investigation of these metabolic pathways as potential therapeutic targets in HCC. In this study, tumor HK2 expression status identified a subset of patients with less favorable prognosis among patients with CKA-positive tumors. In turn, tumor CKA expression identified patients with worse prognosis among patients with HK2-positive tumors. The biologic basis of these observations may be uncertain since the relationship between glucose metabolism and choline metabolism in cancer is not yet well understood. In breast cancer cells, CKA down-regulation has been shown to exert little effect on cellular FDG metabolism, suggesting that glucose metabolism may not be coupled to choline metabolism [47]. This raises the possibility that glucose metabolism and choline metabolism are independently advantageous to tumor cells. A recent PET imaging trial comparing FDG PET with fluorine-18 labeled choline (FC) PET for the detection of HCC reported that welldifferentiated tumors are more likely to be detected by only FC PET, while less differentiated tumors may be detected by either PET technique with similar efficacy [13]. In view of the current study results, it may be hypothesized that livers tumors showing abnormalities on both FDG and FC PET are potentially more aggressive and lethal. One limitation of this study is that the use of microscopy arrays limits the assessment of HK2 and CKA expression to very small portions of tumor. It was not possible to confirm the clinical tumor grade of these samples given the amount of ti.
