The and Huh7-cell lines Transcriptome profiling was performed by next generation sequencing (ProfileXpert, Lyon, France). expression level of is usually inversely correlated to expression, and is associated to poor prognosis for individual SIRT3 survival. To further explore functional effects of the GCK-to-HK2 isoenzyme switch occurring during carcinogenesis, was knocked-out in the HCC cell collection Huh7 and replaced by instead of PP1 vs HCC cell lines provided a unique opportunity to look into HK isoenzyme-dependent metabolic features, lipoprotein production and resistance to immune signals of liver malignancy cells. Results Relative expression level of GCK and HK2 in HCC patients Although an isoenzyme switch from GCK to HK2 has been observed during the carcinogenesis process16, whether hexokinase isoenzymes expression is usually predictive of patient survival is usually unclear. We first analyzed the transcriptomes (RNA-seq data) of 365 HCC biopsies from your Malignancy Genome Atlas (TCGA) database17,18 (Supplementary Data?1). For each HK, the individual gene expression level was used to stratify patients into two subgroups according to Uhlen et al.18 and overall survival in the two subgroups was determined using a Kaplan-Meiers estimator. Although or expression level were not associated to patient survival rate (Fig.?1a), highest expression levels of as previously described19 and least expensive expression levels of in the tumors were associated with a lower survival rate. We thus stratified patients based on the expression ratio to combine these two markers (Fig.?1b). When patients were stratified on PP1 the basis of or expression levels, the median survival between the corresponding subgroups differed by 33.8 and 36.5 months, respectively (Fig.?1a). This difference reached 42.8 months when the stratification of patients was based on the ratio (Fig.?1b). This exhibited that the ratio outperforms or expression alone as predictor of patient survival. Finally, correlation coefficients between patient survival in months and or expression level were decided. For this, we only considered the subset of 130 patients for whom the period between diagnosis and death is usually precisely known (uncensored data), and performed a Spearmans rank correlation test (Fig.?1c). Patient survival was positively correlated to expression but inversely correlated to expression in line with the Kaplan-Meier analysis. In addition, and expression tends to be inversely correlated in tumor samples (Fig.?1c). Therefore, there is a pattern for mutual exclusion of and expression in HCC tumors, and this profile is usually associated to clinical end result. Open in a separate window Fig. 1 Correlation between hexokinase expression levels in HCC tumors and patient survival.a KaplanCMeier estimates of the survival of HCC patients depending on the expression of and (gene expression ratio. The stratification showing the lowest p value when comparing subgroups of patients with the highest to the lowest expression ratio is usually displayed. Patient TCGA-DD-AAE9 exhibiting undetectable levels of and was removed from this analysis as the ratio could not be calculated. c Correlations between patient survival, expression and expression. Spearmans rank correlation test around the subset 130 patients for whom the period between diagnosis and death is usually precisely known (uncensored data). Engineering a cellular model of the hexokinase isoenzyme switch To decipher functional effects of PP1 GCK or HK2 expression in a HCC model, we restored GCK expression by lentiviral transduction in the reference HCC cell collection Huh7, and knocked-out the endogenous gene by CRISPR/Cas9. The.