Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Intestinal dysbacteriosis-induced IL-25 promotes development of HCC via alternative activation of macrophages in tumor microenvironment

doi: 10.1186/s13046-019-1271-3

Figure Lengend Snippet: IL-25 activates alternative macrophages (M2), which promote HCC cell migration and invasion. a IL-25 was used to directly treated HCC cells in vitro, the relative value of CCK-8, Annexin-V, Brdu, Transwell experiment. (Please see results in Additional file : Figure S1A-D). b Immunofluorescent staining was performed in normal liver tissues ( n = 55) and HCC tumor tissues ( n = 98). Representative immunofluorescence images. Bar, 50 μm. c Immunohistochemistry staining was performed in a HCC tissue microarray ( n = 98). Correlation between M2 macrophage (CD206/CD68) percentage and IL-25 scores. CD206 is an M2 macrophage marker, and CD68 is macrophage marker. d and e Macrophages (derived from THP-1) were treated with IL-25 in a time- and concentration-dependent manner. Tumor necrosis factor-α (TNF-α) and CD206 were examined by Western blotting. f and i HCC cells co-cultured with M0 or M2 macrophages. f Cell growth was determined with the Brdu kit. Statistical data were shown at the right. Bar, 100 μm. HCC cells migration ( g ) and invasion ( h ) were determined by Transwell assay. Statistical data were shown at the right. Bar, 100 μm. i Mesenchymal maker vimentin, EMT regulator Snail, epithelial marker E-cadherin, extracellular signal-regulated kinase (ERK), and p-ERK were detected by Western blotting. *** p < 0.001, ns, no significance

Article Snippet: After treatment with IL-25 or co-culture with IL-25-stimulated macrophages, the cell growth, migration and invasion of HCC cell lines (MHCC97L and HepG2) were evaluated by Cell Counting Kit-8 (CCK-8) (Dojindo, Japan), Brdu staining (Sigma-Aldrich) and Transwell assays.

Techniques: Migration, In Vitro, CCK-8 Assay, Staining, Immunofluorescence, Immunohistochemistry, Microarray, Marker, Derivative Assay, Concentration Assay, Western Blot, Cell Culture, Transwell Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Intestinal dysbacteriosis-induced IL-25 promotes development of HCC via alternative activation of macrophages in tumor microenvironment

doi: 10.1186/s13046-019-1271-3

Figure Lengend Snippet: IL-25-induced M2 macrophages promote the EMT process in HCC cells via CXCL10. a - d Macrophages (derived from THP-1) were treated with IL-25 in a time- and concentration-dependent manner. a and b CXCL10 gene expression was quantified by RT-qPCR. c and d CXCL10 protein level was determined by Western blotting. e - g HCC cells were co-cultured with M0 or M2 macrophages. Then, anti-CXCL10 antibody was added to the M2 macrophage culture medium to neutralize CXCL10 protein. Migration ( e ) and invasion ( f ) of HCC cells were determined by Transwell assay. Statistical data were shown at the right. Bar, 100 μm. g Mesenchymal maker vimentin, EMT regulator Snail, epithelial marker E-cadherin, extracellular signal-regulated kinase (ERK), and p-ERK were detected by Western blotting. * p < 0.05, ** p < 0.01, *** p < 0.001, ns, no significance

Article Snippet: After treatment with IL-25 or co-culture with IL-25-stimulated macrophages, the cell growth, migration and invasion of HCC cell lines (MHCC97L and HepG2) were evaluated by Cell Counting Kit-8 (CCK-8) (Dojindo, Japan), Brdu staining (Sigma-Aldrich) and Transwell assays.

Techniques: Derivative Assay, Concentration Assay, Expressing, Quantitative RT-PCR, Western Blot, Cell Culture, Migration, Transwell Assay, Marker

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Intestinal dysbacteriosis-induced IL-25 promotes development of HCC via alternative activation of macrophages in tumor microenvironment

doi: 10.1186/s13046-019-1271-3

Figure Lengend Snippet: IL-25-induced M2 macrophages promote tumorigenesis and EMT of HCC cells in vivo. a Images of tumors from each group. b Tumor weight at the time of sacrifice. c Tumor necrosis factor-α (TNF-α) and CD206 in the tumor tissue of each group were detected by Western blotting. Statistical data were shown at the right. d Mesenchymal maker vimentin, EMT regulator Snail, epithelial marker E-cadherin, extracellular signal-regulated kinase (ERK), and p-ERK were detected in the tumor tissue of each group by Western blotting. Statistical data were shown at the right. e Chemokine CXCL10 was detected in the tumor tissue of each group by Western blotting. Statistical data were shown at the right. * p < 0.05, ** p < 0.01, ns, no significance

Article Snippet: After treatment with IL-25 or co-culture with IL-25-stimulated macrophages, the cell growth, migration and invasion of HCC cell lines (MHCC97L and HepG2) were evaluated by Cell Counting Kit-8 (CCK-8) (Dojindo, Japan), Brdu staining (Sigma-Aldrich) and Transwell assays.

Techniques: In Vivo, Western Blot, Marker

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Intestinal dysbacteriosis-induced IL-25 promotes development of HCC via alternative activation of macrophages in tumor microenvironment

doi: 10.1186/s13046-019-1271-3

Figure Lengend Snippet: Gut bacterial dysbiosis results in colonic epithelial tuft cell hyperplasia and increased secretion of IL-25, which enters the liver via portal vein. IL-25 derived from the gut promotes the alternative activation of macrophages and fosters the tumorigenesis and migration of HCC cells via chemokine CXCL10

Article Snippet: After treatment with IL-25 or co-culture with IL-25-stimulated macrophages, the cell growth, migration and invasion of HCC cell lines (MHCC97L and HepG2) were evaluated by Cell Counting Kit-8 (CCK-8) (Dojindo, Japan), Brdu staining (Sigma-Aldrich) and Transwell assays.

Techniques: Derivative Assay, Activation Assay, Migration

Journal: Molecular Medicine Reports

Article Title: Screening of differentially expressed miRNAs in tensile strain-treated HepG2 cells by miRNA microarray analysis

doi: 10.3892/mmr.2020.11057

Figure Lengend Snippet: BrdU assay results on proliferative ability of HepG2 cells and normal hepatocyte HL-7702 cells. The proliferation ability of HL-7702 cells was not significantly altered after being coated by strain, but that of HepG2 cells was increased. ***P<0.001 vs. control.

Article Snippet: The results showed that type I rat tail collagen coating led to increased cell adherence and growth of HepG2 cells in the BioFlex plates ( ).

Techniques: BrdU Staining, Control

Journal: ACS Omega

Article Title: Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips

doi: 10.1021/acsomega.1c06302

Figure Lengend Snippet: Schematic illustration of DLM micropattern array chips to fabricate HepG2 spheroids for drug screening. DLSs (decellularized liver scaffolds); DLM (decellularized liver matrix); PDMS (poly(dimethylsiloxane)).

Article Snippet: HepG2 cell proliferation was analyzed using the Cell Counting Kit-8 assay (CCK-8, MCE, China).

Techniques:

Journal: ACS Omega

Article Title: Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips

doi: 10.1021/acsomega.1c06302

Figure Lengend Snippet: Preparation of soluble DLM and culture of HepG2 cells on DLM-coated substrates. (A) Powder of DLM. (B) Soluble DLM. (C) Number of HepG2 cells adhesion rate onto each substrate at 4 h after cell seeding. (D) Proliferation of HepG2 cells (cck-8) onto the substrates on day 1 after cell seeding. (E) Viability of HepG2 cells cultured on each substrate ( n = 3). (F) Live/Dead staining of HepG2 cells cultured on each substrate on days 1, 3, and 5. (G) Quantification of albumin secretion from hepatocytes cultured on each substrate using a human albumin ELISA kit ( n = 3). Quantification of urea synthesis by HepG2 cells cultured on each substrate using the urea assay kit ( n = 3). Each substrate (noncoated, DLM, Col I, and Matrigel-coated substrate). * p < 0.05, compared to the noncoated group, # p < 0.05, compared to the DLM-coated group, scale bar = 100 μm.

Article Snippet: HepG2 cell proliferation was analyzed using the Cell Counting Kit-8 assay (CCK-8, MCE, China).

Techniques: CCK-8 Assay, Cell Culture, Staining, Enzyme-linked Immunosorbent Assay

Journal: ACS Omega

Article Title: Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips

doi: 10.1021/acsomega.1c06302

Figure Lengend Snippet: Fabrication of the round DLM micropattern array chips and the formation of 3D HepG2 cellular aggregates. (A) Round micropattern of PDMS seals with different diameters. (B) Proper homogeneous transfer of DLM fluorescently labeled to create round DLM micropattern array chips with different diameters. (C) Microscopic images of HepG2 cells cultured on round micropattern array chips with different diameters at 6 h. (D) Microscopic images of cellular aggregates cultured on round micropattern array chips with different diameters on day 3. Different diameters (50, 75, 100, 150, 200, and 300 μm). Scale bar = 100 μm.

Article Snippet: HepG2 cell proliferation was analyzed using the Cell Counting Kit-8 assay (CCK-8, MCE, China).

Techniques: Labeling, Cell Culture

Journal: ACS Omega

Article Title: Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips

doi: 10.1021/acsomega.1c06302

Figure Lengend Snippet: Evaluation of 3D HepG2 cellular aggregates of different sizes. (A) 3D view of cellular aggregates on the round DLM micropattern. (B) Live/Dead staining of cellular aggregates with different diameters cultured on round DLM micropattern array chips. Dead cells are stained red, while viable cells are stained green. Scale bar = 50 μm. (C) Gene expression levels of hepatic genes alb , aat , ck 18, cyp 1 a 1, mrp 2, and tat were determined for different diameters compared to cells cultured on tissue culture plastic (TCP). *p <0.05, compared to the 2D, #p < 0.05, compared to the 100 μm, not significant (ns), compared to the 100 μm. Different diameters (50, 75, 100, 150, 200, and 300 μm).

Article Snippet: HepG2 cell proliferation was analyzed using the Cell Counting Kit-8 assay (CCK-8, MCE, China).

Techniques: Staining, Cell Culture, Expressing

Journal: ACS Omega

Article Title: Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips

doi: 10.1021/acsomega.1c06302

Figure Lengend Snippet: HepG2 spheroids cultured on DLM micropattern array chips (100 μm round micropattern) for anticancer drug screening. (A) Live/Dead staining of HepG2 spheroids before being treated with DMSO or drugs. (B) Live/Dead staining of HepG2 spheroids with various concentrations (5–100 μg/mL) of paclitaxel, doxorubicin HCl, and disulfiram. Scale bar = 100 μm.

Article Snippet: HepG2 cell proliferation was analyzed using the Cell Counting Kit-8 assay (CCK-8, MCE, China).

Techniques: Cell Culture, Staining

Journal: ACS Omega

Article Title: Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips

doi: 10.1021/acsomega.1c06302

Figure Lengend Snippet: Quantitatively analyzing the efficacy of an anticancer drug. The viability of HepG2 spheroids (mean gray value) with various concentrations (5–100 μg/mL) of paclitaxel, doxorubicin HCl, and disulfiram at (A) 24 h and (B) 48 h and the viability of HepG2 spheroids (CCK-8) with various concentrations (5–100 μg/mL) of paclitaxel, doxorubicin HCl, and disulfiram at (C) 24 h and (D) 48 h ( *p < 0.05, compared to doxorubicin HCl). (E) Correlation between mean gray value and viability detected by CCK-8 ( r indicates correlation coefficient).

Article Snippet: HepG2 cell proliferation was analyzed using the Cell Counting Kit-8 assay (CCK-8, MCE, China).

Techniques: CCK-8 Assay

Journal: Oncology Letters

Article Title: Metformin attenuates hypoxia-induced resistance to cisplatin in the HepG2 cell line

doi: 10.3892/ol.2018.9869

Figure Lengend Snippet: The influence of hypoxia on survival and cell growth in the HepG2 liver cancer cell line. (A) HepG2 cells showed time-dependent growth in normoxic conditions, but proliferation was significantly inhibited in hypoxic conditions, with a significant difference at 48 h compared to normoxia (P=0.005). (B) HepG2 cells showed G1 arrest in hypoxic conditions. The results represent the mean ± SE of three independent experiments; **P<0.01 vs. normoxic conditions, as determined by Student's t-test for each time point (A), or the cell cycle distribution at 0 h, as determined by one-way ANOVA with Turkey-Kramer's post-hoc test (B). ANOVA, one-way analysis of variance.

Article Snippet: Cell culture and growth conditions The human liver cancer cell line HepG2, the human lung cancer cell line A549, and the human oral squamous carcinoma cell line SAS were purchased from the Riken BioResource Center (Tsukuba, Ibaraki, Japan).

Techniques:

Journal: Oncology Letters

Article Title: Metformin attenuates hypoxia-induced resistance to cisplatin in the HepG2 cell line

doi: 10.3892/ol.2018.9869

Figure Lengend Snippet: Hypoxia inhibits induction of apoptosis by cisplatin in HepG2 cells. Flow cytograms of cells harvested from cultures after treatment with cisplatin were analyzed. (A) Of the 7-AAD-negative population in these gated cells, a fraction of Annexin V-positive cells was induced by cisplatin in normoxic conditions. Apoptosis was induced by cisplatin in normoxic conditions at 48 h, but the change was not clear in hypoxic conditions. (B and C) These values were expressed as a percentage. Induction of apoptosis by cisplatin increased in a dose-dependent manner in normoxic conditions (P<0.05 at 5 µM at 24 h and P<0.01 at 1 and 5 µM at 48 h), but this effect was attenuated at 48 h in hypoxic conditions. The results represent the mean ± SE of three independent experiments; *P<0.05 and **P<0.01 vs. untreated cells for each oxygen condition, as determined by one-way ANOVA by following Dunnett's test. ††P<0.01 vs. normoxic conditions, as determined by Student's t-test for each concentration. CDDP, cisplatin; FITC, Fluorescein isothiocyanate; 7-AAD, 7-Aminoactinomycin D.

Article Snippet: Cell culture and growth conditions The human liver cancer cell line HepG2, the human lung cancer cell line A549, and the human oral squamous carcinoma cell line SAS were purchased from the Riken BioResource Center (Tsukuba, Ibaraki, Japan).

Techniques: Concentration Assay

Journal: Oncology Letters

Article Title: Metformin attenuates hypoxia-induced resistance to cisplatin in the HepG2 cell line

doi: 10.3892/ol.2018.9869

Figure Lengend Snippet: Attenuation of hypoxia-induced cisplatin resistance by a non-toxic concentration of metformin. (A) Metformin lower than 2 mM did not inhibit the proliferation of HepG2 cells in either normoxic or hypoxic conditions. The results represent the mean ± SE of three independent experiments as determined by one-way ANOVA by following Dunnett's test. (B) In normoxia, cisplatin showed the same cytotoxicity regardless of the presence or absence of metformin. (C) In hypoxia, the cytotoxicity of cisplatin was increased by 1 mM metformin. (D and E) Combination treatment increased apoptosis compared to treatment with cisplatin alone in hypoxic conditions. The results represent the mean ± SE of three independent experiments; *P<0.05 and **P<0.01 vs. cisplatin alone, as determined by Student's t-test or Welch's t-test. CDDP, cisplatin; ANOVA, one-way analysis of variance.

Article Snippet: Cell culture and growth conditions The human liver cancer cell line HepG2, the human lung cancer cell line A549, and the human oral squamous carcinoma cell line SAS were purchased from the Riken BioResource Center (Tsukuba, Ibaraki, Japan).

Techniques: Concentration Assay

Journal: Oncology Letters

Article Title: Metformin attenuates hypoxia-induced resistance to cisplatin in the HepG2 cell line

doi: 10.3892/ol.2018.9869

Figure Lengend Snippet: Hypoxia-induced factor 1α mediates a pathway in which metformin modulates hypoxia-induced resistance to cisplatin. (A) HepG2 cells expressed HIF-1α in hypoxic conditions in the absence of metformin. Metformin suppressed the expression of HIF-1α. Metformin did not activate AMPKα or AMPKβ, but activated Akt and mTOR. This change was remarkable at 24 h. (B) siRNA significantly inhibited HIF-1α gene expression. The effect of combining metformin were less modulated in siHIF-treated cells. (C) sicontrol: P=0.134, (D) siHIF; P=0.585; *P<0.05. CDDP, cisplatin; HIF, hypoxia-inducible factor; AMPK, adenosine monophosphate-activated protein kinase; p-, phosphorylated; si, small interfering; mTOR, mammalian target of rapamycin.

Article Snippet: Cell culture and growth conditions The human liver cancer cell line HepG2, the human lung cancer cell line A549, and the human oral squamous carcinoma cell line SAS were purchased from the Riken BioResource Center (Tsukuba, Ibaraki, Japan).

Techniques: Expressing, Gene Expression