human hepatocellular carcinoma cell line  (ATCC)

Journal: International Journal of Hepatology

Article Title: LOX‐1 and Its Splicing Variant LOX‐1 Δ 4 Differently Influence Hepatocellular Carcinoma Cell Progression and Aggressiveness

doi: 10.1155/ijh/5704085

Figure Lengend Snippet: LOX‐1 expression increases in poorly differentiated hepatocellular carcinoma (HCC). (A) Representative immunohistochemical images of LOX‐1 expression in control liver and non‐HCV/HBV‐related HCCs tissues at 10x and 40x magnification (insert), scale bars = 50 μ m. G3 HCC also displays a nuclear localization of LOX‐1 compared with the only cytoplasmic distribution of the other experimental groups. (B) Semiquantitative analysis of immunostaining demonstrates a significant increase of LOX‐1 expression in G2 and G3 HCC tissue compared with G1 ones and control liver. Results are represented as mean ± SEM. Statistical significance was assessed by one‐way ANOVA followed by Bonferroni post hoc test: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Human hepatoblastoma line from a 15‐year‐old male, commonly known as HepG2 (RRID:CVCL_0027), was used as a model of well‐differentiated epithelial neoplastic transformation that arises following chronic, non‐HCV/HBV‐related inflammatory stages, obtained from the American Type Culture Collection (ATCC, Manassas, Virginia, United States) [ ].

Techniques: Expressing, Immunohistochemical staining, Control, Analysis, Immunostaining, Tissue

Journal: International Journal of Hepatology

Article Title: LOX‐1 and Its Splicing Variant LOX‐1 Δ 4 Differently Influence Hepatocellular Carcinoma Cell Progression and Aggressiveness

doi: 10.1155/ijh/5704085

Figure Lengend Snippet: LOX‐1 Δ 4 overexpression increases proliferation and chemoresistance in transfected HepG2 cells. (A) Proliferation rate of LOX‐1 and LOX‐1 Δ 4‐overexpressing HepG2 cells compared with control cells (empty) after 48 and 72 h from transient transfection. Results are represented as mean ± SEM. Statistical significance was assessed by two‐way ANOVA followed by Bonferroni post hoc test: ∗ p < 0.05 and ∗∗ p < 0.01. (B) Cell viability (CCK8 assay) of 48 h‐transfected cells treated and not with additional 24 h of Doxo (1 μ M, Doxo). Data are normalized and expressed as percentage (%) relative to the respective untreated control (set to 100%) as mean ± SEM of three independent experiments. Statistical significance for chemoresistance protection was assessed by Student′s t ‐test: asterisk (∗) indicates p < 0.05 when comparing Doxo‐treated LOX‐1 Δ 4‐overexpressing HepG2 cells versus Doxo‐treated empty vector‐transfected cells. (C) Western blot analysis of nuclear and cytoplasmic protein expression of p21 Cip1/Waf1 , p27 Kip1 and acetyl‐histone H4. SP1 and GAPDH were utilized as specific loading and purity controls for the nuclear and cytoplasmic fractions, respectively. (D) Densitometric analysis of p21 Cip1/Waf1 , p27 Kip1 and acetyl‐histone H4 expression. Results are represented as mean ± SEM. Statistical significance among the three experimental groups was assessed by one‐way ANOVA followed by Bonferroni post hoc test: ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus empty and LOX‐1 transfected‐HepG2 cells.

Article Snippet: Human hepatoblastoma line from a 15‐year‐old male, commonly known as HepG2 (RRID:CVCL_0027), was used as a model of well‐differentiated epithelial neoplastic transformation that arises following chronic, non‐HCV/HBV‐related inflammatory stages, obtained from the American Type Culture Collection (ATCC, Manassas, Virginia, United States) [ ].

Techniques: Over Expression, Transfection, Control, CCK-8 Assay, Plasmid Preparation, Western Blot, Analysis, Expressing

Journal: International Journal of Hepatology

Article Title: LOX‐1 and Its Splicing Variant LOX‐1 Δ 4 Differently Influence Hepatocellular Carcinoma Cell Progression and Aggressiveness

doi: 10.1155/ijh/5704085

Figure Lengend Snippet: LOX‐1 Δ 4 overexpression differentially modulates Cyclin D1 and p53 expression in transfected HepG2 cells. (A) Representative immunofluorescence images of HepG2 cells 72 h posttransfection with control cells (empty), LOX‐1 and LOX‐1 Δ 4 splicing variant. Nuclei are counterstained with Hoechst (blue, top row). Panels show the intracellular distribution and expression levels of Cyclin D1 (red, second row) and p53 (green, third row), with the merged overlay demonstrating compartmental colocalization patterns (bottom row). Scale bar = 25 μ m. (B) Digital quantification analysis of average fluorescence intensities for Cyclin D1 and p53 within the respective subcellular compartments, validating the distinct downregulation of p53 and upregulation of Cyclin D1 selectively driven by LOX‐1 Δ 4 isoform. Data are expressed as mean ± SEM. Statistical significance was assessed by one‐way ANOVA followed by Bonferroni post hoc test: ∗ p < 0.05 and ∗∗ p < 0.01 versus empty and LOX‐1.

Article Snippet: Human hepatoblastoma line from a 15‐year‐old male, commonly known as HepG2 (RRID:CVCL_0027), was used as a model of well‐differentiated epithelial neoplastic transformation that arises following chronic, non‐HCV/HBV‐related inflammatory stages, obtained from the American Type Culture Collection (ATCC, Manassas, Virginia, United States) [ ].

Techniques: Over Expression, Expressing, Transfection, Immunofluorescence, Control, Variant Assay, Analysis, Fluorescence

Journal: International Journal of Hepatology

Article Title: LOX‐1 and Its Splicing Variant LOX‐1 Δ 4 Differently Influence Hepatocellular Carcinoma Cell Progression and Aggressiveness

doi: 10.1155/ijh/5704085

Figure Lengend Snippet: Expression analysis of autophagy‐ and EMT‐related markers in LOX‐1 and LOX‐1 Δ 4 transfected HepG2 cells. (A) Representative images of immunoblotting for p62/SQSTM1 and LC3B markers. The analysis documented a prominent expression of the lipidated LC3B‐II form, whereas the cytosolic LC3B‐I pool remained below the threshold of densitometric detection. (B,C) Densitometric analysis of p62/SQSTM1 and LC3B expression. (D) Western blot analysis of E‐cadherin and vimentin markers. (E,F) Densitometric analysis of E‐cadherin and vimentin expression. Results are represented as mean ± SEM. Statistical significance was assessed by one‐way ANOVA followed by Bonferroni post hoc test: ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: Human hepatoblastoma line from a 15‐year‐old male, commonly known as HepG2 (RRID:CVCL_0027), was used as a model of well‐differentiated epithelial neoplastic transformation that arises following chronic, non‐HCV/HBV‐related inflammatory stages, obtained from the American Type Culture Collection (ATCC, Manassas, Virginia, United States) [ ].

Techniques: Expressing, Analysis, Transfection, Western Blot

Journal: Chemmedchem

Article Title: Scaffold Hopping Provides Liver X Receptor Agonist With Preference for the β Subtype

doi: 10.1002/cmdc.70509

Figure Lengend Snippet: In vitro characterization of the optimized LXR agonist 7 . (a) Full‐length LXR reporter gene assay in HEK293T cells. N ≥ 3. (b) Quantification of LXR target genes in HepG2 cells. Both 2 and ( S )‐ 7 robustly induced expression of target genes involved in cholesterol metabolism (ABCA1, ABCG1) and DNL (FAS, SREBP1c). Data are the mean ± SEM of N = 4; ** p < 0.01, *** p < 0.001 (ANOVA with post hoc Bonferroni test against 0.1% DMSO or as indicated by the bars). Full agonist T0901317 ( 32 ) was tested for comparison. (c,d) Lipogenesis assay. Incubation of HepG2 cells with pan LXR full agonist T0901317 ( 32 ) for 72 h led to a substantial accumulation of lipids that were stained with Oil Red O. In comparison to 32 , both the parent compound 2 and ( S )‐ 7 displayed less lipogenic potential. Data are the mean ± SEM of N = 3; * p < 0.05, ** p < 0.01, *** p < 0.001 (one‐sided t ‐test against 0.1% DMSO). The images shown in (d) are representative details from the full images that can be found in Figure S3. (e) In a cytotoxicity assay, 2 and ( rac )‐ 7 did not influence the metabolic activity of HepG2 cells but induced cytotoxicity to HEK293T cells in the micromolar concentration region. While these concentrations are well above the EC 50 value of ( S )‐ 7 in the Gal4 reporter gene assay (red dashed line), compound 2 showed partially toxic effects at its EC 50 value (green dashed line). (f) Metabolic stability of 2 and ( rac )‐ 7 in rat liver microsomes. After 60 min, ( rac )‐ 7 was almost recovered completely, whereas approximately half of 2 had been metabolized. 7‐Ethoxycoumarine (7‐EC) served as internal standard. Data are the mean ± SEM of N = 3.

Article Snippet: HEK293T and HepG2 cells (German Collection of Microorganisms and Cell Cultures, DSMZ) were grown in Dulbecco's modified Eagle's medium (DMEM) high glucose supplemented with 10% fetal bovine serum (FBS), sodium pyruvate (1 mM), penicillin (100 U/mL), and streptomycin (100 μg/mL) at 37 °C and 5% CO 2 .

Techniques: In Vitro, Reporter Gene Assay, Expressing, Comparison, Incubation, Staining, Cytotoxicity Assay, Activity Assay, Concentration Assay, Stability

Journal: Food Science & Nutrition

Article Title: Protective Effects of Nongxiang‐Type Tieguanyin on the Liver of D‐Galactose‐Induced Aging C57BL /6 Mice via the NF ‐ κB Signaling Pathway

doi: 10.1002/fsn3.72396

Figure Lengend Snippet: Tgy‐N reduces cellular senescence markers in Nutlin‐3a‐induced senescent AML12 and HepG2 cells. (A) SA‐β‐gal staining of AML12 cells. Scale bar = 100 μm. (B) Quantitative analysis of SA‐β‐gal‐positive AML12 cells. (C) Representative Western blots of p16, p21, and GAPDH in AML12 cells. (D) Quantitative analysis of p16/GAPDH ratio in AML12 cells. (E) Quantitative analysis of p21/GAPDH ratio in AML12 cells. (F) SA‐β‐gal staining of HepG2 cells. Scale bar = 100 μm. (G) Quantitative analysis of SA‐β‐gal‐positive HepG2 cells. (H) Representative Western blots of p16, p21, and GAPDH in HepG2 cells. (I) Quantitative analysis of p16/GAPDH ratio in HepG2 cells. (J) Quantitative analysis of p21/GAPDH ratio in HepG2 cells. Data are expressed as mean ± SEM. Statistical assessments were performed using one‐way ANOVA followed by post hoc Duncan's test. Different letters above the bars denote statistically significant differences between groups at the p < 0.05 level.

Article Snippet: AML12 (mouse normal hepatocytes) and HepG2 (human hepatocellular carcinoma) cell lines were obtained from ATCC.

Techniques: Staining, Analysis, Western Blot