Journal: Cancers

Article Title: Transcriptional Control of Hepatocellular Carcinoma Cells Aggressiveness by AAV2/8-Mediated Delivery of Human Centenarian-Associated SIRT6 N308K/A313S

doi: 10.3390/cancers18050812

Figure Lengend Snippet: Titration of AAV2/8 Luc in HepG2 and Huh-7 liver cancer cells. Three multiplicities of infection (MOI) doses (10 4 , 10 5 , 10 6 —viral genomes) were tested in HepG2 ( upper panel) and in Huh-7 liver cancer cells ( lower panel) at three time points (24 h, 48 h, 72 h). Untransduced cells (CTL) were used as negative controls. Data are presented as mean ± SEM, n = 3.

Article Snippet: HepG2 and Huh-7 liver cancer cells were obtained from CLS-GmbH (Eppelheim, Germany) and were authenticated using STR profiling.

Techniques: Titration, Infection

Journal: Cancers

Article Title: Transcriptional Control of Hepatocellular Carcinoma Cells Aggressiveness by AAV2/8-Mediated Delivery of Human Centenarian-Associated SIRT6 N308K/A313S

doi: 10.3390/cancers18050812

Figure Lengend Snippet: mRNA expression of SIRT6 WT or SIRT6 Cent in HepG2 ( A ) and in Huh-7 ( B ) liver cancer cells. Cells were transduced with AAV2/8-Luc, AAV2/8-SIRT6-WT and AAV2/8-SIRT6-N308K/A313S for 48 h. mRNA levels of SIRT6 WT and SIRT6c were assessed by qPCR. Data are presented as mean ± SEM, n = 3. p < 0.001 (***) versus AAV2/8-Luc and AAV2/8-SIRT6-N308K/A313S-transduced cells.

Article Snippet: HepG2 and Huh-7 liver cancer cells were obtained from CLS-GmbH (Eppelheim, Germany) and were authenticated using STR profiling.

Techniques: Expressing, Transduction

Journal: Cancers

Article Title: Transcriptional Control of Hepatocellular Carcinoma Cells Aggressiveness by AAV2/8-Mediated Delivery of Human Centenarian-Associated SIRT6 N308K/A313S

doi: 10.3390/cancers18050812

Figure Lengend Snippet: MTT viability assays: MTT assay on HepG2 ( A ); and Huh-7 liver cancer cell line ( B ) transduced with AAV2/8-Luc, AAV2/8-SIRT6-WT and AAV2/8-SIRT6-N308K/A313S for up to five days (0 h, 24 h, 48 h, 72 h, 96 h, 120 h). Cell survivals were significantly lower in AAV2/8-SIRT6-WT and AAV2/8-SIRT6-N308K/A313S-transduced cells from AAV2-8-Luc at the indicated time points, with a p < 0.05 (*), p < 0.01 (**) or p < 0.001 (***). Data are presented as mean ± SEM, n = 3.

Article Snippet: HepG2 and Huh-7 liver cancer cells were obtained from CLS-GmbH (Eppelheim, Germany) and were authenticated using STR profiling.

Techniques: MTT Assay, Transduction

Journal: Cancers

Article Title: Transcriptional Control of Hepatocellular Carcinoma Cells Aggressiveness by AAV2/8-Mediated Delivery of Human Centenarian-Associated SIRT6 N308K/A313S

doi: 10.3390/cancers18050812

Figure Lengend Snippet: Global transcriptomic profiling in hepatocellular carcinoma cells: two-dimensional principal component analysis (PCA) plots derived from variance-stabilized expression values (DESeq2) for HepG2 ( A ); and Huh7 ( B ) liver cancer cells. Each point represents an individual biological replicate, illustrating clear separation of samples according to experimental condition (Luc control, SIRT6-WT, and SIRT6-Cent), reflecting distinct overall gene-expression signatures.

Article Snippet: HepG2 and Huh-7 liver cancer cells were obtained from CLS-GmbH (Eppelheim, Germany) and were authenticated using STR profiling.

Techniques: Derivative Assay, Expressing, Control, Gene Expression

Journal: Cancers

Article Title: Transcriptional Control of Hepatocellular Carcinoma Cells Aggressiveness by AAV2/8-Mediated Delivery of Human Centenarian-Associated SIRT6 N308K/A313S

doi: 10.3390/cancers18050812

Figure Lengend Snippet: Differential expression and pathway analysis in HepG2 liver cancer cells: ( A ) volcano plots displaying differentially expressed genes across the three pairwise comparisons (SIRT6-WT vs. Luc, SIRT6-Cent vs. Luc, and SIRT6-Cent vs. SIRT6-WT). The horizontal axis shows log 2 fold-change, and the vertical axis indicates −log 10 (adjusted p -value). Upregulated genes are shown in blue, downregulated genes in red; and ( B ) reactome pathway enrichment results for the same comparisons. The bubble size corresponds to the number of differentially expressed genes contributing to each pathway, while color intensity reflects the degree of statistical enrichment.

Article Snippet: HepG2 and Huh-7 liver cancer cells were obtained from CLS-GmbH (Eppelheim, Germany) and were authenticated using STR profiling.

Techniques: Quantitative Proteomics

Journal: Cancers

Article Title: Transcriptional Control of Hepatocellular Carcinoma Cells Aggressiveness by AAV2/8-Mediated Delivery of Human Centenarian-Associated SIRT6 N308K/A313S

doi: 10.3390/cancers18050812

Figure Lengend Snippet: Invasive capacity of HCC cells after SIRT6 overexpression: invasion through basement membrane-coated inserts was evaluated in HepG2 ( A ); and Huh7 ( B ) liver cancer cells 48 h after transduction with AAV2/8-Luc (control), AAV2/8-SIRT6-WT, or AAV2/8-SIRT6-N308K/A313S, followed by a 24 h migration period toward 10% FBS as chemoattractant. White bars: no chemoattractant (negative control); black bars: migration inducer provided with the kit (positive control). Both SIRT6-WT and the centenarian-associated SIRT6 variant dramatically reduced cell invasion compared with the Luc control. Data are presented as mean ± SEM, n = 3. Data are presented as mean ± SEM, n = 3. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001 versus AAV2/8-Luc; $$$ p < 0.001 versus AAV2/8-SIRT6-WT.

Article Snippet: HepG2 and Huh-7 liver cancer cells were obtained from CLS-GmbH (Eppelheim, Germany) and were authenticated using STR profiling.

Techniques: Over Expression, Membrane, Transduction, Control, Migration, Negative Control, Positive Control, Variant Assay

Journal: Cancers

Article Title: Transcriptional Control of Hepatocellular Carcinoma Cells Aggressiveness by AAV2/8-Mediated Delivery of Human Centenarian-Associated SIRT6 N308K/A313S

doi: 10.3390/cancers18050812

Figure Lengend Snippet: Nanoindentation assays in: HepG2 ( A ); and Huh-7 liver cancer cell lines ( B ) transduced with AAV2/8-Luc, AAV2/8-SIRT6-WT and AAV2/8-SIRT6-N308K/A313S for two days prior to incubation in the Boyden chamber for a further 24 h; 10% FBS was used as chemoattractant. Cell stiffness (measured in Pascal, Pa) was significantly higher in AAV2/8- AAV2/8-SIRT6-N308K/A313S-transduced cells compared to AAV2/8-Luc and to AAV2/8-SIRT6-WT. p < 0.05 (*), p < 0.01 (**) versus AAV2/8-SIRT6-WT.

Article Snippet: HepG2 and Huh-7 liver cancer cells were obtained from CLS-GmbH (Eppelheim, Germany) and were authenticated using STR profiling.

Techniques: Transduction, Incubation