human normal hepatocyte cell line hepatocytes  (ATCC)

Journal: Translational Cancer Research

Article Title: Analysis of the role of POC1A in the development and progression of hepatocellular carcinoma

doi: 10.21037/tcr-23-2398

Figure Lengend Snippet: Analysis of the expression of POC1A in various classifications of HCC patients, as well as in HCC and liver cancer cell lines. (A) Comparison of POC1A between cancer tissue samples and normal tissue samples. (B) Comparison of the expression of POC1A in different genders. (C) Comparison of the expression of POC1A in different age groups. (D) The expression levels of POC1A were all higher in the Caucasian, African-American, and Asian HCC patients than the normal patients. The expression of POC1A was significantly higher in LIHC tissue samples from Asian patients than LIHC tissue samples from Caucasian patients. (E) The expression of POC1A in individual cancer stages. (F) The expression of POC1A in different transistor grades. (G) The expression of POC1A in tumor (T) stage. (H) The expression of POC1A in node (N) stage. (I) The expression of POC1A in metastasis (M) stage. (J) Analysis of POC1A expression in normal liver tissue and cancer tissue samples from patients with hepatocellular carcinoma using immunohistochemical methods in the Human Protein Atlas database. Liver sample: https://www.proteinatlas.org/ENSG00000164087-POC1A/tissue/liver#img , hepatocellular samples: https://www.proteinatlas.org/ENSG00000164087-POC1A/pathology/liver+cancer#img . Scale bar: 100 µm. (K) Analysis of POC1A mRNA expression in LO2 and HepG2 cell lines by qPCR assay. *, P<0.05; **, P<0.01; -: no significance. TCGA, The Cancer Genome Atlas; HCC, hepatocellular carcinoma; LIHC, liver hepatocellular carcinoma; mRNA, messenger RNA; qPCR, quantitative polymerase chain reaction; POC1A , POC1 centriolar protein A.

Article Snippet: The human normal hepatocyte cell line LO2 (Servicebio Technology Co., Ltd., Wuhan, China) was cultured in Roswell Park Memorial Institute 1640 medium, and the hepatoma cell line HepG2 (Procell Life Science & Technology Co., Ltd., Wuhan, China) was cultured in Dulbecco’s Modified Eagle Medium (Hyclone, Thermo Fisher Scientific Inc., Waltham, MA), supplemented with 10% fetal bovine serum and antibiotics (10,000 U/mL penicillin and 10 mg/mL streptomycin) (Procell Life Science & Technology Co., Ltd.).

Techniques: Expressing, Comparison, Immunohistochemical staining, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: G protein–coupled estrogen receptor 1 ameliorates nonalcoholic steatohepatitis through targeting AMPK-dependent signaling

doi: 10.1016/j.jbc.2024.105661

Figure Lengend Snippet: GPER1 plays a crucial role in the protective effects of estradiol on lipid accumulation, oxidative stress, and inflammation in L02 cells under metabolic stress. A , representative images of Nile Red staining in L02 cells treated with vehicle or GPER1-specific agonist G1 (100 nM) followed by BSA or PO (palmitic acid and oil acid mixture) stimulation for 12 h (n = 3 independent experiments). Scale bar represents 100 μm. B , triglyceride (TG) and total cholesterol (TC) contents in L02 cells from the indicated group (n = 6). C , relative mRNA levels of factors related to fatty acid metabolism in L02 cells from the indicated group (n = 6). D , representative images of DCFH-DA probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. E , representative images of MitoSOX Red probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. F , relative mRNA levels of factors related to inflammatory response in L02 cells from the indicated group (n = 6). G , representative images of Nile Red staining of L02 cells challenged by PO and treated with vehicle, 17β-estradiol (E2; 10 nM), GPER1 antagonist G15 (10 μM), or E2 in combination with G15 (n = 3 independent experiments). Scale bar represents 100 μm. H , TG and TC contents in L02 cells from the indicated group (n = 6). I , relative mRNA levels of factors related to fatty acid metabolism in L02 cells from the indicated group (n = 6). J , representative images of DCFH-DA probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. K , representative images of MitoSOX Red probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. L , relative mRNA levels of factors related to inflammatory response in L02 cells from the indicated group (n = 6). M , immunoblotting analysis of GPER1 protein level in the WT and GPER1 KO L02 cells (n = 3 independent experiments). N , representative images of Nile Red staining in the WT and GPER1 KO L02 cells challenged by PO and cotreated with vehicle or E2 (n = 3 independent experiments). Scale bar represents 100 μm. O , TG and TC contents in L02 cells from the indicated group (n = 6). P , relative mRNA levels of factors related to fatty acid metabolism in L02 cells from the indicated group (n = 6). Q , representative images of DCFH-DA probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. R , representative images of MitoSOX Red probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. S , relative mRNA levels of factors related to inflammatory response in L02 cells from the indicated group (n = 6). In all statistical plots, data are expressed as the mean ± SD and analyzed by one-way ANOVA with Bonferroni analysis. ∗∗∗ p < 0.001, comparison between the indicated groups; n.s., no significance, p ≥ 0.05, comparison between the indicated groups. The mRNA expression of target genes was normalized to that of Actb. BSA, bovine serum albumin; GPER1, G protein–coupled estrogen receptor 1; PO, palmitic acid-oleic acid mixture.

Article Snippet: The L02 cell lines (human normal hepatocyte) were purchased from the China Center for Type Culture Collection and cultured in Roswell Park Memorial Institute 1640 medium with 10% FBS and 1% penicillin-streptomycin.

Techniques: Staining, Western Blot, Comparison, Expressing

Journal: The Journal of Biological Chemistry

Article Title: G protein–coupled estrogen receptor 1 ameliorates nonalcoholic steatohepatitis through targeting AMPK-dependent signaling

doi: 10.1016/j.jbc.2024.105661

Figure Lengend Snippet: GPER1 mediates the release of cAMP and activates the AMPK signaling pathway. A , cyclic AMP levels in PO (palmitic acid and oil acid mixture)-challenged L02 cells for 30 min after treatment with 10 −10 to 10 −5 M GPER1-specific agonist G1 (n = 4). B , cyclic AMP levels in PO-challenged L02 cells after treatment with G1 (100 nM) for the indicated times (n = 4). C , cyclic AMP levels in WT or GPER1 KO L02 cells challenged by PO and cotreated with vehicle or 17β-estradiol (E2; 10 nM) for the indicated times (n = 3). D , cyclic AMP levels in WT or GPER1 KO HepG2 cells were challenged by PO and cotreated with vehicle or E2 for the indicated times (n = 3). E , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in primary hepatocytes that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of Gαs inhibitor NF449 (10 μM). F , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ) and primary hepatocytes ( right ) that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of AC inhibitor MDL-12330A (20 μM). G , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ), HepG2 cells ( middle ), and primary hepatocytes ( right ) that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of EPAC antagonist ESI-09 (10 μM). H , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ), HepG2 cells ( middle ), and primary hepatocytes ( right ) that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of PKA inhibitor H89 (10 μM). I , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ) and HepG2 cells ( right ) challenged by PO and cotreated with vehicle or G1 in the absence or presence of PKA catalytic subunit α (PKAc) siRNA. In all statistical plots, data are expressed as the mean ± SD and analyzed by one-way ANOVA with Bonferroni analysis. For ( A ) and ( B ), ∗ p < 0.05, ∗∗∗ p < 0.001, G1-PO group versus Vehicle-PO group. For ( C ) and ( D ), ∗∗∗ p < 0.001, comparison between the indicated groups; n.s., no significance, p ≥ 0.05, comparison between the indicated groups. AC, adenylyl cyclase; AMPK, AMP-activated protein kinase; EPAC, exchange protein activated by cAMP; GPER1, G protein–coupled estrogen receptor 1; PO, palmitic acid-oleic acid mixture.

Article Snippet: The L02 cell lines (human normal hepatocyte) were purchased from the China Center for Type Culture Collection and cultured in Roswell Park Memorial Institute 1640 medium with 10% FBS and 1% penicillin-streptomycin.

Techniques: Western Blot, Comparison