Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39.

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet: Figure 2. Loss of ARG1 and AGMAT enhances liver tumor formation (A) Immunoblots of arginine-to-polyamine-converting enzymes (ARG1 and AGMAT) and polyamine metabolism enzymes (ODC, SRM, SMS, SAT1, PAOX, and SMOX) in Ctrl liver and L-dKO tumor tissues. Calnexin serves as loading control (same samples were used as in Figure 1E). n = 4 (Ctrl), n = 8 (L-dKO). (B) Total polyamine content in Ctrl liver and L-dKO tumor tissues. n = 6. (C) Relative 3H-putrescine uptake into Ctrl liver and L-dKO tumor tissues. n = 8. (D) Immunohistochemistry of Ctrl and L-dKO liver tissues stained for ARG1 or AGMAT. NT, adjacent non-tumor tissue; T, tumor. (E) Representative images of livers from L-dKO mice injected with AAV-Ctrl, AAV-ARG1, or AAV-AGMAT. (F) Number of macroscopic tumors per liver of L-dKO mice injected with AAV-Ctrl, AAV-ARG1, or AAV-AGMAT. n = 9–10. (G) Arginine content in Ctrl liver and L-dKO non-tumor (NT) and tumor (T) tissues of mice injected with AAV-Ctrl, AAV-ARG1, or AAV-AGMAT. n = 4–10. *p < 0.05, **p < 0.01. ***p < 0.001, ****p < 0.0001 by unpaired t test (B and C) and one-way ANOVA (F and G).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER L-Arginine-coupled agarose beads gbiosciences Cat#GENO786-1361 L-leucine-coupled agarose beads gbiosciences Cat#GENO786-1370 Critical commercial assays Total Polyamine Assay Kit BioVision Cat#K475 L-arginine ELISA kit MyBiosource Cat#MBS728648-96 Nano-Glo dual luciferase reporter assay kit Promega Cat#N1610 Deposited data HCC patients’ RNA sequencing data Ng et al., 202250 EGAS00001005073, EGAS00001005074 HCC patients’ proteome data Ng et al., 202250 PRIDE (PXD025705, PXD025836) Mouse RNA sequencing data Dimitrakopoulos et al., 202125 SRP156216 Mouse proteome data Dimitrakopoulos et al., 202125 https://github.com/cbgethz/netics/tree/ master/mouse_data Mouse and HCC patients’ metabolomics This paper MASSIVE: MSV000092406 RNA sequencing data of ARG1/AGMAT-expressing (and control) and RBM39-depleted (and control) SNU-449 cells This paper GEO: PRJNA940402 Mouse, HCC patients, and SNU-449 proteome data of pulldowns using Leu- and Arg-coupled agarose beads This paper MASSIVE: MSV000091516 Experimental models: Cell lines SNU-182 Diego Calvisi (University of Sassari, Italy) N/A SNU-449 Diego Calvisi (University of Sassari, Italy) N/A HLE Diego Calvisi (University of Sassari, Italy) N/A PLC Diego Calvisi (University of Sassari, Italy) N/A Hep40 Diego Calvisi (University of Sassari, Italy) N/A Hep3B Diego Calvisi (University of Sassari, Italy) N/A Huh6 Diego Calvisi (University of Sassari, Italy) N/A Huh7 Diego Calvisi (University of Sassari, Italy) N/A HepG2 Diego Calvisi (University of Sassari, Italy) N/A SNU-475 Gerhard Christofori (University of Basel, Switzerland) N/A SNU-423 Gerhard Christofori (University of Basel, Switzerland) N/A Huh1 Gerhard Christofori (University of Basel, Switzerland) N/A HEK293 ATCC CRL-1573 HEK239T ATCC CRL-3216 SNU-449 control This paper N/A SNU-449 ARG1 This paper N/A SNU-449 AGMAT This paper N/A SNU-449 ARG1/AGMAT This paper N/A SNU-449 ASS1-FLAG This paper N/A SNU-449 ARG1/AGMAT +ASS1 This paper N/A SNU-449 ARG1/AGMAT +3xHA-ASS1 This paper N/A Huh7 sgCtrl This paper N/A Huh7 sgARG1 This paper N/A Huh7 sgAGMAT This paper N/A SNU-449 ARG1/AGMAT +control (for ASNS) This paper N/A (Continued on next page) ll OPEN ACCESS Cell 186, 5068–5083.e1–e12, November 9, 2023 e2 Article

Techniques: Western Blot, Control, Immunohistochemistry, Staining, Injection

Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39.

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet: Figure 3. ARG1/AGMAT determine metabolic gene expression via arginine (A) Immunoblots of SNU-449 cells upon stable expression of ARG1 and/or AGMAT. Actin serves as loading control. (B) Representative clonogenic growth assay of control, ARG1-, and/or AGMAT-expressing SNU-449 cells grown in arginine-restricted medium. (C) Relative clonogenic growth of control, ARG1-, and/or AGMAT- expressing SNU-449 cells. N = 6. (D) Arginine content of control, ARG1-, and/or AGMAT-expressing SNU-449 cells. N = 4. (E) PCA analysis of RNA-seq data of control and ARG1/AGMAT-expressing SNU-449 cells. (F) Heatmap of a subset of differentially expressed metabolic genes in ARG1/AGMAT-expressing compared to control SNU-449 cells (log2 fold-change). (G) mRNA levels of ASNS, PSAT1, PSPH, GLSK, GLUT3, HK2, NNMT, and AOC3 in control and ARG1/AGMAT-expressing SNU-449 cells. N = 5–7. (H) Immunoblots of ASNS, PSAT, PSPH, and NNMT from two independent experiments of control and ARG1/AGMAT-expressing SNU-449 cells. Calnexin serves as loading control. (I) Immunoblots of ASNS, PSAT, PSPH, and NNMT of Ctrl liver and L-dKO tumor tissues. Calnexin serves as loading control. n = 4 (Ctrl), n = 8 (L-dKO). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA (C and D) and unpaired t test (G).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER L-Arginine-coupled agarose beads gbiosciences Cat#GENO786-1361 L-leucine-coupled agarose beads gbiosciences Cat#GENO786-1370 Critical commercial assays Total Polyamine Assay Kit BioVision Cat#K475 L-arginine ELISA kit MyBiosource Cat#MBS728648-96 Nano-Glo dual luciferase reporter assay kit Promega Cat#N1610 Deposited data HCC patients’ RNA sequencing data Ng et al., 202250 EGAS00001005073, EGAS00001005074 HCC patients’ proteome data Ng et al., 202250 PRIDE (PXD025705, PXD025836) Mouse RNA sequencing data Dimitrakopoulos et al., 202125 SRP156216 Mouse proteome data Dimitrakopoulos et al., 202125 https://github.com/cbgethz/netics/tree/ master/mouse_data Mouse and HCC patients’ metabolomics This paper MASSIVE: MSV000092406 RNA sequencing data of ARG1/AGMAT-expressing (and control) and RBM39-depleted (and control) SNU-449 cells This paper GEO: PRJNA940402 Mouse, HCC patients, and SNU-449 proteome data of pulldowns using Leu- and Arg-coupled agarose beads This paper MASSIVE: MSV000091516 Experimental models: Cell lines SNU-182 Diego Calvisi (University of Sassari, Italy) N/A SNU-449 Diego Calvisi (University of Sassari, Italy) N/A HLE Diego Calvisi (University of Sassari, Italy) N/A PLC Diego Calvisi (University of Sassari, Italy) N/A Hep40 Diego Calvisi (University of Sassari, Italy) N/A Hep3B Diego Calvisi (University of Sassari, Italy) N/A Huh6 Diego Calvisi (University of Sassari, Italy) N/A Huh7 Diego Calvisi (University of Sassari, Italy) N/A HepG2 Diego Calvisi (University of Sassari, Italy) N/A SNU-475 Gerhard Christofori (University of Basel, Switzerland) N/A SNU-423 Gerhard Christofori (University of Basel, Switzerland) N/A Huh1 Gerhard Christofori (University of Basel, Switzerland) N/A HEK293 ATCC CRL-1573 HEK239T ATCC CRL-3216 SNU-449 control This paper N/A SNU-449 ARG1 This paper N/A SNU-449 AGMAT This paper N/A SNU-449 ARG1/AGMAT This paper N/A SNU-449 ASS1-FLAG This paper N/A SNU-449 ARG1/AGMAT +ASS1 This paper N/A SNU-449 ARG1/AGMAT +3xHA-ASS1 This paper N/A Huh7 sgCtrl This paper N/A Huh7 sgARG1 This paper N/A Huh7 sgAGMAT This paper N/A SNU-449 ARG1/AGMAT +control (for ASNS) This paper N/A (Continued on next page) ll OPEN ACCESS Cell 186, 5068–5083.e1–e12, November 9, 2023 e2 Article

Techniques: Gene Expression, Western Blot, Expressing, Control, Growth Assay, RNA Sequencing

Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39.

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet: Figure 4. ASNS promotes arginine uptake in liver cancer (A) Relative 3H-arginine uptake in control and ARG1/AGMAT-expressing SNU-449 cells with or without pre-loading with asparagine (Asn) or glutamine (Gln). N = 5–6. (B) Immunoblots of ARG1/AGMAT-expressing SNU-449 cells upon stable expression of ASNS or control. Calnexin serves as loading control. (C) Relative 3H-arginine uptake in control and ASNS-expressing SNU-449 ARG1/AGMAT-ex- pressing cells. N = 5. (D) Representative clonogenic growth assay of control and ASNS-expressing SNU-449 ARG1/ AGMAT-expressing cells grown in arginine- restricted medium. (E) mRNA levels of PSAT1, PSPH, GLSK, GLUT3, HK2, NNMT, and AOC3 in control and ASNS- expressing SNU-449 ARG1/AGMAT-expressing cells. N = 6–8. (F) Immunoblots of ASNS, PSAT, PSPH, and NNMT from two independent experiments of control and ASNS-expressing SNU-449 ARG1/ AGMAT-expressing cells. Calnexin serves as loading control. (G) mRNA levels of Asns in L-dKO non-tumor (NT) and tumor (T) tissues of mice injected with AAV- shCtrl or AAV-shAsns. n = 6–7. (H) Number of macroscopic tumors per liver in L-dKO mice injected with AAV-shCtrl or AAV- shAsns. n = 7. (I) Arginine content in L-dKO non-tumor (NT) and tumor (T) tissues of mice injected with AAV-shCtrl or AAV-shAsns. n = 4–6. n.s. = not significant; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by unpaired t test (A, C, E, G, and H) and one-way ANOVA (I).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER L-Arginine-coupled agarose beads gbiosciences Cat#GENO786-1361 L-leucine-coupled agarose beads gbiosciences Cat#GENO786-1370 Critical commercial assays Total Polyamine Assay Kit BioVision Cat#K475 L-arginine ELISA kit MyBiosource Cat#MBS728648-96 Nano-Glo dual luciferase reporter assay kit Promega Cat#N1610 Deposited data HCC patients’ RNA sequencing data Ng et al., 202250 EGAS00001005073, EGAS00001005074 HCC patients’ proteome data Ng et al., 202250 PRIDE (PXD025705, PXD025836) Mouse RNA sequencing data Dimitrakopoulos et al., 202125 SRP156216 Mouse proteome data Dimitrakopoulos et al., 202125 https://github.com/cbgethz/netics/tree/ master/mouse_data Mouse and HCC patients’ metabolomics This paper MASSIVE: MSV000092406 RNA sequencing data of ARG1/AGMAT-expressing (and control) and RBM39-depleted (and control) SNU-449 cells This paper GEO: PRJNA940402 Mouse, HCC patients, and SNU-449 proteome data of pulldowns using Leu- and Arg-coupled agarose beads This paper MASSIVE: MSV000091516 Experimental models: Cell lines SNU-182 Diego Calvisi (University of Sassari, Italy) N/A SNU-449 Diego Calvisi (University of Sassari, Italy) N/A HLE Diego Calvisi (University of Sassari, Italy) N/A PLC Diego Calvisi (University of Sassari, Italy) N/A Hep40 Diego Calvisi (University of Sassari, Italy) N/A Hep3B Diego Calvisi (University of Sassari, Italy) N/A Huh6 Diego Calvisi (University of Sassari, Italy) N/A Huh7 Diego Calvisi (University of Sassari, Italy) N/A HepG2 Diego Calvisi (University of Sassari, Italy) N/A SNU-475 Gerhard Christofori (University of Basel, Switzerland) N/A SNU-423 Gerhard Christofori (University of Basel, Switzerland) N/A Huh1 Gerhard Christofori (University of Basel, Switzerland) N/A HEK293 ATCC CRL-1573 HEK239T ATCC CRL-3216 SNU-449 control This paper N/A SNU-449 ARG1 This paper N/A SNU-449 AGMAT This paper N/A SNU-449 ARG1/AGMAT This paper N/A SNU-449 ASS1-FLAG This paper N/A SNU-449 ARG1/AGMAT +ASS1 This paper N/A SNU-449 ARG1/AGMAT +3xHA-ASS1 This paper N/A Huh7 sgCtrl This paper N/A Huh7 sgARG1 This paper N/A Huh7 sgAGMAT This paper N/A SNU-449 ARG1/AGMAT +control (for ASNS) This paper N/A (Continued on next page) ll OPEN ACCESS Cell 186, 5068–5083.e1–e12, November 9, 2023 e2 Article

Techniques: Control, Expressing, Western Blot, Growth Assay, Injection

Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39.

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet: Figure 7. ARG1, AGMAT, arginine, and RBM39 in human HCC patients (A) Schematic representation of arginine and polyamine metabolism in HCC patients. Boxes below enzymes indicate changes in mRNA (left box) and protein (right box) levels in human HCC tumors (T) compared to paired non-tumor (NT) biopsies, respectively. Color coding according to level of log2 fold-change as indicated. ‘‘?’’ indicates unknown identity. Tumor aggressiveness is indicated by Edmondson-Steiner grade low (Edm. low, grade I and II) and high (Edm. high, grade III and IV). n = 73 (Edm. low) and n = 49 (Edm. high) for mRNA; n = 30 (Edm. low) and n = 21 (Edm. high) for protein. (B) Immunoblots of ARG1, AGMAT, RBM39, and ASNS in paired non-tumor (NT) and tumor (T) tissues of five HCC patients. Calnexin serves as loading control. (C) Tissue microarray for ARG1 and AGMAT. ARG1, normal liver n = 58, HCC n = 160; AGMAT, normal liver n = 49, HCC n = 142. (D) Representative IHC of ARG1 and AGMAT of an HCC patient (from C). Non-tumor, NT; tumor, T.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER L-Arginine-coupled agarose beads gbiosciences Cat#GENO786-1361 L-leucine-coupled agarose beads gbiosciences Cat#GENO786-1370 Critical commercial assays Total Polyamine Assay Kit BioVision Cat#K475 L-arginine ELISA kit MyBiosource Cat#MBS728648-96 Nano-Glo dual luciferase reporter assay kit Promega Cat#N1610 Deposited data HCC patients’ RNA sequencing data Ng et al., 202250 EGAS00001005073, EGAS00001005074 HCC patients’ proteome data Ng et al., 202250 PRIDE (PXD025705, PXD025836) Mouse RNA sequencing data Dimitrakopoulos et al., 202125 SRP156216 Mouse proteome data Dimitrakopoulos et al., 202125 https://github.com/cbgethz/netics/tree/ master/mouse_data Mouse and HCC patients’ metabolomics This paper MASSIVE: MSV000092406 RNA sequencing data of ARG1/AGMAT-expressing (and control) and RBM39-depleted (and control) SNU-449 cells This paper GEO: PRJNA940402 Mouse, HCC patients, and SNU-449 proteome data of pulldowns using Leu- and Arg-coupled agarose beads This paper MASSIVE: MSV000091516 Experimental models: Cell lines SNU-182 Diego Calvisi (University of Sassari, Italy) N/A SNU-449 Diego Calvisi (University of Sassari, Italy) N/A HLE Diego Calvisi (University of Sassari, Italy) N/A PLC Diego Calvisi (University of Sassari, Italy) N/A Hep40 Diego Calvisi (University of Sassari, Italy) N/A Hep3B Diego Calvisi (University of Sassari, Italy) N/A Huh6 Diego Calvisi (University of Sassari, Italy) N/A Huh7 Diego Calvisi (University of Sassari, Italy) N/A HepG2 Diego Calvisi (University of Sassari, Italy) N/A SNU-475 Gerhard Christofori (University of Basel, Switzerland) N/A SNU-423 Gerhard Christofori (University of Basel, Switzerland) N/A Huh1 Gerhard Christofori (University of Basel, Switzerland) N/A HEK293 ATCC CRL-1573 HEK239T ATCC CRL-3216 SNU-449 control This paper N/A SNU-449 ARG1 This paper N/A SNU-449 AGMAT This paper N/A SNU-449 ARG1/AGMAT This paper N/A SNU-449 ASS1-FLAG This paper N/A SNU-449 ARG1/AGMAT +ASS1 This paper N/A SNU-449 ARG1/AGMAT +3xHA-ASS1 This paper N/A Huh7 sgCtrl This paper N/A Huh7 sgARG1 This paper N/A Huh7 sgAGMAT This paper N/A SNU-449 ARG1/AGMAT +control (for ASNS) This paper N/A (Continued on next page) ll OPEN ACCESS Cell 186, 5068–5083.e1–e12, November 9, 2023 e2 Article

Techniques: Western Blot, Control, Microarray

Journal: International journal of molecular sciences

Article Title: eIF4A1 Is a Prognostic Marker and Actionable Target in Human Hepatocellular Carcinoma.

doi: 10.3390/ijms24032055

Figure Lengend Snippet: Figure 5. Human hepatocellular carcinoma cell lines express eIF4A1, eIF4A2, and DDX3 proteins. (A) The levels of the three proteins were assessed in HLE, HLF, PLF/PRF/5, SNU182, and SNU449 cell lines by western blot analysis. GAPDH was used as a loading control, and protein band intensities of eIF4A1, eIF4A2, and DDX3 were normalized to GAPDH levels (B).

Article Snippet: The human HCC cell lines HLE (JCRB0404, Xenotech, Kansas City, MO, USA), HLF (JCRB0405, Tebu-bio GmbH, Offenbach, Germany), PLC/PRF/5 (300315, CLS Cell lines Service GmbH, Eppelheim, Germany), SNU182 (CRL-2235, LGC Standards GmbH), and SNU449 (CRL-2234, LGC Standards GmbH) were cultured at 37 ◦C in a 5% CO2 humidified atmosphere.

Techniques: Western Blot, Control

Journal: International journal of molecular sciences

Article Title: eIF4A1 Is a Prognostic Marker and Actionable Target in Human Hepatocellular Carcinoma.

doi: 10.3390/ijms24032055

Figure Lengend Snippet: Figure 6. Cell viability of human hepatocellular carcinoma cell lines exposed to eIF4A inhibitors, as assessed by MTT assay. The eIF4A1 inhibitors, FL3, Zotatifin, CR-1-31-B, Rocaglamide, and Silvestrol, were administered to HLE (A), HLF (B), and PLC/PRF/5 (C) cell lines. Similar data were obtained in the SNU449 cell line (not shown). Data of two independent experiments with n = 4 technical replicates are represented as the percentage of DMSO-treated cells ± SD. A summary of the IC50 ± SD of the five drugs in the three cell lines is depicted in (D).

Article Snippet: The human HCC cell lines HLE (JCRB0404, Xenotech, Kansas City, MO, USA), HLF (JCRB0405, Tebu-bio GmbH, Offenbach, Germany), PLC/PRF/5 (300315, CLS Cell lines Service GmbH, Eppelheim, Germany), SNU182 (CRL-2235, LGC Standards GmbH), and SNU449 (CRL-2234, LGC Standards GmbH) were cultured at 37 ◦C in a 5% CO2 humidified atmosphere.

Techniques: MTT Assay

Journal: Cancer letters

Article Title: B cell CLL/lymphoma 6 member B inhibits hepatocellular carcinoma metastases in vitro and in mice.

doi: 10.1016/j.canlet.2014.08.025

Figure Lengend Snippet: Fig. 1. Transcriptional silencing of BCL6B in HCC is associated with DNA methylation. (A) BCL6B expression was determined by RT-PCR. BCL6B was silenced in many HCC cell lines (upper left). Methylation of BCL6B was determined by methylation specific PCR (MSP) (lower left) and bisulfite sequencing (BGS) (right). (B) BCL6B mRNA expres- sion was determined by quantitative real-time PCR in 40 pairs of early (TNM stage I/II) and advanced (TNM stage III/IV) HCC. A significant reduction in BCL6B mRNA expression was observed in advanced HCC tumors as compared with their adjacent normal tissues. (C) The protein expression of BCL6B in HCC cell lines and normal liver tissues was determined by western blot. (D) The mRNA expression of BCL6B was restored after treatment with demethylation agent 5-Aza.

Article Snippet: Human HCC cell lines and tissue samples Ten human HCC cell lines (HepG2, Hep3B, PLC-5, Huh1, Huh6, Huh7, Bel7404, SNU398, SNU449 and MHCC97L) were obtained from ATCC.

Techniques: DNA Methylation Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Methylation, Sequencing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cancer letters

Article Title: B cell CLL/lymphoma 6 member B inhibits hepatocellular carcinoma metastases in vitro and in mice.

doi: 10.1016/j.canlet.2014.08.025

Figure Lengend Snippet: Fig. 2. BCL6B overexpression inhibits the growth of HCC cells. (A) Ectopic expression of BCL6B in Huh7 and HepG2 cell lines was confirmed by RT-PCR and western blot. (B) Cell viability was reduced by overexpression of BCL6B in Huh7 and HepG2 cells. (C) Ectopic expression of BCL6B suppressed colony formation.

Article Snippet: Human HCC cell lines and tissue samples Ten human HCC cell lines (HepG2, Hep3B, PLC-5, Huh1, Huh6, Huh7, Bel7404, SNU398, SNU449 and MHCC97L) were obtained from ATCC.

Techniques: Over Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Cancer letters

Article Title: B cell CLL/lymphoma 6 member B inhibits hepatocellular carcinoma metastases in vitro and in mice.

doi: 10.1016/j.canlet.2014.08.025

Figure Lengend Snippet: Fig. 3. BCL6B induced cell apoptosis and inhibited tumor growth in nude mice. (A) BCL6B induced apoptosis in Huh7 and HepG2 cells, as determined by flow cytometry analysis following Annexin V and 7-AAD staining. The upper panel represents FACS images of HCC cells transfected with empty vector or BCL6B, while the lower panel shows the quantitative analyses of early apoptotic and late apoptotic cells. The experiment was repeated three times in triplicate. Data are mean ± SD. (B) Protein expres- sion of the active forms of apoptosis related genes caspase 3, caspase 7, and poly (ADP-ribose) polymerase (PARP) was evaluated by Western blot. GAPDH was used as a loading control. (C) Ectopic expression of BCL6B inhibited the growth of subcutaneous HepG2 xenografts in nude mice.

Article Snippet: Human HCC cell lines and tissue samples Ten human HCC cell lines (HepG2, Hep3B, PLC-5, Huh1, Huh6, Huh7, Bel7404, SNU398, SNU449 and MHCC97L) were obtained from ATCC.

Techniques: Cytometry, Staining, Transfection, Plasmid Preparation, Western Blot, Control, Expressing

Journal: Cancer letters

Article Title: B cell CLL/lymphoma 6 member B inhibits hepatocellular carcinoma metastases in vitro and in mice.

doi: 10.1016/j.canlet.2014.08.025

Figure Lengend Snippet: Fig. 4. Effect of BCL6B on HCC cell migration and invasion. (A) Representative result of scratch healing assay. (B) Representative result of invasion assay. The experiment was repeated three times in triplicate. Data are mean ± SD.

Article Snippet: Human HCC cell lines and tissue samples Ten human HCC cell lines (HepG2, Hep3B, PLC-5, Huh1, Huh6, Huh7, Bel7404, SNU398, SNU449 and MHCC97L) were obtained from ATCC.

Techniques: Migration, Invasion Assay

Journal: Cancer letters

Article Title: B cell CLL/lymphoma 6 member B inhibits hepatocellular carcinoma metastases in vitro and in mice.

doi: 10.1016/j.canlet.2014.08.025

Figure Lengend Snippet: Fig. 6. Enhanced expression of BCL6B suppressed HCC metastasis to lung in vivo. (A) Tumors derived from the luciferase-labelled MHCC97L cells were successfully im- planted orthotopically into the livers of nude mice and tumors can be detected in all the mice after 8 weeks. Tumor growth in the mice was monitored by a live imaging system. (B) Lung metastasis of the HCC mice with or without BCL6B overexpression at 8 weeks after orthotopic implantation. The lung metastatic nodules were analyzed histologically. Quantitative analyses showed the lung metastasis incidence in Lenti-BCL6B group was significantly lower than that in the Lenti-Vector group. (C) Western blots were performed to validate the downstream gene expression regulated by BCL6B in HepG2, Huh7 and MHCC97L. GAPDH was used as an internal control.

Article Snippet: Human HCC cell lines and tissue samples Ten human HCC cell lines (HepG2, Hep3B, PLC-5, Huh1, Huh6, Huh7, Bel7404, SNU398, SNU449 and MHCC97L) were obtained from ATCC.

Techniques: Expressing, In Vivo, Derivative Assay, Luciferase, Imaging, Over Expression, Plasmid Preparation, Western Blot, Gene Expression, Control