Journal: PLoS ONE

Article Title: Switching of Pyruvate Kinase Isoform L to M2 Promotes Metabolic Reprogramming in Hepatocarcinogenesis

doi: 10.1371/journal.pone.0115036

Figure Lengend Snippet: MiR-122 targeted PKM2 and suppressed PKM2 expression. (A) Seed sequence of miR-122 in the 3UTR of PKM2 was underlined. WT and mutated sequences of PKM2 were inserted into pmiRGLO luciferase vector. (B) Re-expression of miR-122 suppressed luciferase activity of the WT but not the Mut 3UTR of PKM2. EV and sensor sequences served as negative and positive controls, respectively. (C) Re-expression of miR-122 in SMMC-7221 and MHCC-97L cells suppressed PKM2 but not PKL/R expression. (D) Linear regression model demonstrated that PKM2 mRNA expression was inversely correlated with miR-122 expression in HCC (pink) and NT liver tissues (blue). (E) Expression levels of 667 miRNA species in HCC and NT were plotted. Each dot represents one individual miRNA. MiR-122 is the most abundant miRNA in NT liver. (F) MiR-122 expression in NT, HCC, and venous metastases (VM). Data were retrieved from low density microarray in which expressions of 667 miRNA species were compared between NT, HCC, and VM. B , * P

Article Snippet: Antibodies, HCC tissue microarray sections, immunohistochemistry and histology PKM1 (Sigma), phospho-PKM2 (Tyr105) (Cell Signaling Technology, Danvers, MA), PKM2 (Cell Signaling Technology), PKL (Abcam), β actin (Sigma) were used for Western Blotting.

Techniques: Expressing, Sequencing, Luciferase, Plasmid Preparation, Activity Assay, Microarray

Journal: PLoS ONE

Article Title: Switching of Pyruvate Kinase Isoform L to M2 Promotes Metabolic Reprogramming in Hepatocarcinogenesis

doi: 10.1371/journal.pone.0115036

Figure Lengend Snippet: PKM2 expression in human HCC. (A) mRNA expression of PKL, PKM1, and PKM2 in HCC and NT tissues. Values = 2 ΔΔCT , ΔΔCT = (CT PK – CT HPRT ) of HCC - (CT PK – CT HPRT ) of NT. P values, Wilcoxin signed rank test (B) Waterfall plot shows that, at the mRNA level, PKM2 was up-regulated (HCC/NT2 folds) in 29/60 (48.33%) human HCC samples. (C) Representative pictures of IHC staining with antibody against PKM2 in HCC tissue microarray. PKM2 protein was drastically up-regulated in human HCCs as compared to the paired NT tissues. (D) Mann Whitney test showed that PKM2 over-expression was associated with multiple aggressive clinicopathological features in HCC including the presence of tumor microsatellites, presence of venous invasion, and absence of tumor encapsulation. (E) Over-expression of PKM2 in human HCC was associated with poor prognosis. HCC patients were categorized into two groups: PKM2 over-expression and PKM2 normal/under-expression. PKM2 was considered to be over-expressed when HCC/NT2 folds and was considered to be normal/under-expressed otherwise. HCC patients with PKM2 over-expression had a higher 1-year tumor recurrence rate after surgical resection than HCC patients without PKM2 over-expression, 46.667% Vs 25%. (F) Patients with PKM2 over-expression had lower 5-year overall survival rates after surgical resection. P values were calculated by Kaplan-Meir log rank test.

Article Snippet: Antibodies, HCC tissue microarray sections, immunohistochemistry and histology PKM1 (Sigma), phospho-PKM2 (Tyr105) (Cell Signaling Technology, Danvers, MA), PKM2 (Cell Signaling Technology), PKL (Abcam), β actin (Sigma) were used for Western Blotting.

Techniques: Expressing, Immunohistochemistry, Staining, Microarray, MANN-WHITNEY, Over Expression