Journal: Aging Cell

Article Title: Liver osteopontin is required to prevent the progression of age‐related nonalcoholic fatty liver disease

doi: 10.1111/acel.13183

Figure Lengend Snippet: Liver OPN is p53 regulated. (a) Protein levels of p53 were evaluated by immunohistochemistry in liver sections of 3‐, 10‐, and 20‐month‐old (m) wild‐type (WT) mice ( n = 3–5). (b) OPN protein levels from liver homogenates were measured in WT and p53‐KO male and female mice fed a chow diet (CD) and a high‐fat diet (HFD) by immunoblotting using glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) as loading control ( n = 4–6). (c) Liver p53 and OPN protein levels were evaluated in WT mice fed a HFD injected with p53‐dominant positive adenovirus (adp53) and GFP (adGFP), using transferrin or GAPDH as a loading control ( n = 4–6). (d) Protein levels of p53 and OPN in siCtrl and sip53 HepG2 cells were measured by immunoblotting using GAPDH as loading control. Extracellular OPN was measured using an ELISA ( n = 4–5). (e) OPN protein levels from HepG2 cells silenced for p53 and Hep3B cells treated with palbociclib (Palbo) were measured by immunoblotting using GAPDH as loading control. OPN media levels from Hep3B cells treated either with vehicle (Veh) or with palbociclib (Palbo) were measured by ELISA ( n = 4–8). Values are means ± SEM . Significant differences are denoted by * p < 0.05, ** p < 0.01, and *** p < 0.001 (Student's t test)

Article Snippet: Data obtained from the Cell Line Encyclopedia (© 2019 The Broad Institute of MIT & Harvard) showed that the cell line with the least p53 expression, the Hep3B, also had the lowest OPN expression.

Techniques: Immunohistochemistry, Western Blot, Control, Injection, Enzyme-linked Immunosorbent Assay

Journal: Archives of Medical Science : AMS

Article Title: MicroRNA-490-3p suppresses hepatocellular carcinoma cell proliferation and migration by targeting the aurora kinase A gene ( AURKA )

doi: 10.5114/aoms.2019.91351

Figure Lengend Snippet: miR-490-3p suppresses expression of aurora kinase A gene ( AURKA ) in hepatocellular carcinoma (HCC) cells. A – qRT-PCR showed that miR-390-3p mimic transfection could significantly increase expression of miR-390-3p in HepG2 cells. B – qRT-PCR showed that miR-390-3p mimic transfection could significantly increase expression of miR- 390-3p in HepG2 cells. C – qRT-PCR showed that AURKA mRNA expression of miR-490-3p mimics and AURKA siRNA group was significantly lower, while AURKA mRNA expression of AURKA cDNA group was higher, and miR-490-3p + AURKA cDNA had no influence on AURKA mRNA expression in HepG2 cells. D – qRT-PCR showed that AURKA mRNA expression of miR-490-3p mimics and AURKA siRNA group was significantly lower, while AURKA mRNA expression of AURKA cDNA group was higher, and miR-490-3p + AURKA cDNA had no influence on the AURKA mRNA expression in Hep3B cells. E – Western blot showed that AURKA protein expression of miR-490-3p mimics and AURKA siRNA group was significantly lower, while AURKA protein expression of AURKA cDNA group was higher, and miR-490-3p + AURKA cDNA had no influence on AURKA protein expression in HepG2 cells. F – Western blot showed that AURKA protein expression of miR-490-3p mimics and AURKA siRNA group was significantly lower, while AURKA protein expression of AURKA cDNA group was higher, and miR-490-3p + AURKA cDNA had no influence on AURKA protein expression in Hep3B cells *p < 0.05, **p < 0.01, compared with negative control (NC) group.

Article Snippet: Human embryonic kidney cell line HEK-293T, human HCC cell lines Huh-7, HepG2, Hep3B and SK-HEP1 and normal liver cell line HL-7702 were purchased from the Japanese Collection of Research Bioresources (Tokyo, Japan).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Negative Control

Journal: Archives of Medical Science : AMS

Article Title: MicroRNA-490-3p suppresses hepatocellular carcinoma cell proliferation and migration by targeting the aurora kinase A gene ( AURKA )

doi: 10.5114/aoms.2019.91351

Figure Lengend Snippet: Impact of miR-490-3p and aurora kinase A gene ( AURKA ) on proliferation of hepatocellular carcinoma (HCC) cells. A – Colony formation images of HepG2 and Hep3B cells showed that colony number of miR-490-3p mimics and AURKA siRNA group was significantly smaller, while colony number area of AURKA cDNA group was larger, and miR-490-3p + AURKA cDNA had no influence on proliferation of the cells. B – Statistical graph of colony formation of HepG2 cells. C – Statistical graph of colony formation of Hep3B cells. D – MTT assay showed that the cell viability of miR-490-3p mimics and AURKA siRNA group was significantly lower, while the cell viability area of AURKA cDNA group was higher, and miR-490-3p + AURKA cDNA had no influence on HepG2 cell viability. E – MTT assay showed that cell viability of miR-490-3p mimics and AURKA siRNA group was significantly lower, while cell viability area of AURKA cDNA group was higher, and miR-490-3p + AURKA cDNA had no influence on Hep3B cell viability *p < 0.05, **p < 0.01, compared with negative control (NC) group.

Article Snippet: Human embryonic kidney cell line HEK-293T, human HCC cell lines Huh-7, HepG2, Hep3B and SK-HEP1 and normal liver cell line HL-7702 were purchased from the Japanese Collection of Research Bioresources (Tokyo, Japan).

Techniques: MTT Assay, Negative Control

Journal: Archives of Medical Science : AMS

Article Title: MicroRNA-490-3p suppresses hepatocellular carcinoma cell proliferation and migration by targeting the aurora kinase A gene ( AURKA )

doi: 10.5114/aoms.2019.91351

Figure Lengend Snippet: Impact of miR-490-3p and aurora kinase A gene ( AURKA ) on migration of hepatocellular carcinoma cells. A – Wound healing images of HepG2 and Hep3B cells showed that closed wound area of miR-490-3p mimics and AURKA siRNA group was significantly smaller, while closed wound area of AURKA cDNA group was larger, and miR-490-3p + AURKA cDNA had no influence on migration of the cells. B – Statistical graph of closed wound area of HepG2 cells. C – Statistical graph of closed wound area of Hep3B cells *p < 0.05, **p < 0.01, compared with negative control (NC) group.

Article Snippet: Human embryonic kidney cell line HEK-293T, human HCC cell lines Huh-7, HepG2, Hep3B and SK-HEP1 and normal liver cell line HL-7702 were purchased from the Japanese Collection of Research Bioresources (Tokyo, Japan).

Techniques: Migration, Negative Control

Journal: Archives of Medical Science : AMS

Article Title: MicroRNA-490-3p suppresses hepatocellular carcinoma cell proliferation and migration by targeting the aurora kinase A gene ( AURKA )

doi: 10.5114/aoms.2019.91351

Figure Lengend Snippet: Impact of miR-490-3p and aurora kinase A gene ( AURKA ) on invasion of hepatocellular carcinoma cells. A – Transwell images of HepG2 and Hep3B cells showed that invasive cell number of miR-490-3p mimics and AURKA siRNA group was significantly decreased, while invasive cell number of AURKA cDNA group was increased, and miR-490-3p + AURKA cDNA had no influence on invasion of the cells. B – Statistical graph of invasive cell number of HepG2 cells. C – Statistical graph of invasive cell number of Hep3B cells *p < 0.05, **p < 0.01, compared with negative control (NC) group.

Article Snippet: Human embryonic kidney cell line HEK-293T, human HCC cell lines Huh-7, HepG2, Hep3B and SK-HEP1 and normal liver cell line HL-7702 were purchased from the Japanese Collection of Research Bioresources (Tokyo, Japan).

Techniques: Negative Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Antioxidant and Anti-Cancer Potentials of Rheum emodi Rhizome Extracts

doi: 10.1093/ecam/neq048

Figure Lengend Snippet: IC 50 values of aqueous and methanolic extracts of R. emodi in MDA-MB-435S and Hep3B cell lines as determined in XTT assay.

Article Snippet: MDA-MB-435S (human breast carcinoma) and Hep3B (human liver carcinoma) cell lines were obtained from National Centre for Cell Science (Pune, India).

Techniques:

Journal: BMC Cancer

Article Title: Dysregulated paired related homeobox 1 impacts on hepatocellular carcinoma phenotypes

doi: 10.1186/s12885-021-08637-3

Figure Lengend Snippet: PRRX1 and metabolism of HCC cells. A Glucose consumption and lactate output were measured 48 h after siPRRX1 transfection. Bars indicate mean ± SD and a representative picture of 3 experiments is shown, each in triplicate. B Expression of genes involved in glycolysis were determined by qPCR 48 h after siPRRX1 transfection in HUH7 and HLF cells. Bars indicate mean ± SD and representative of 3 experiments each in triplicates. C Expression of genes involved in the TCA cycle were analyzed by qPCR 48 h after siPRRX1 transfection in HUH7 and HLF cells. Bars indicate mean ± SD and representative of 3 experiments, each in triplicates. D Expression of genes involved in amino acid metabolism as determined by qPCR 48 h after siPRRX1 transfection in HUH7 and HLF cells. Bars indicate mean ± SD and representative of 3 experiments each in triplicates. E Alterations of metabolites. Heatmap of changes in metabolite levels of glycolysis, TCA cycle and amino acids in HUH7 and HLF cells 48 h after siPRRX1 transfection. n = 3 per cell line and condition. White numbers indicate a p < 0.05; black numbers are not significant. The experiment was performed in triplicate

Article Snippet: The cell lines HLF, HUH7, Hep3B and SNU398 were cultured in Dulbecco Modified Eagle’s medium (DMEM, high glucose, Lonza, BE12–709) supplemented with 2 mM glutamine, 10% fetal bovine serum (FBS), penicillin (100 U/ml), and streptomycin (100 μg/ml).

Techniques: Transfection, Expressing

Journal: BMC Cancer

Article Title: Dysregulated paired related homeobox 1 impacts on hepatocellular carcinoma phenotypes

doi: 10.1186/s12885-021-08637-3

Figure Lengend Snippet: Summary of metabolic changes after knockdown of PRRX1 ( A ) in HUH7 and ( B ) in HLF cells. Red indicates upregulation, green indicates downregulation of genes or metabolites, italics: gene expression, bold: dysregulated metabolites. See text for details

Article Snippet: The cell lines HLF, HUH7, Hep3B and SNU398 were cultured in Dulbecco Modified Eagle’s medium (DMEM, high glucose, Lonza, BE12–709) supplemented with 2 mM glutamine, 10% fetal bovine serum (FBS), penicillin (100 U/ml), and streptomycin (100 μg/ml).

Techniques: Knockdown, Gene Expression

Journal: World Journal of Gastroenterology

Article Title: Exosome-mediated transfer of circRNA563 promoting hepatocellular carcinoma by targeting the microRNA148a-3p/metal-regulatory transcription factor-1 pathway

doi: 10.3748/wjg.v29.i46.6060

Figure Lengend Snippet: Effects of mesenchymal stem cells or mesenchymal stem cell-derived exosome co-incubation on Hep3B cells. A-C: The proliferation of Hep3B cells was measured using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and colony formation assays; D-G: Invasion and metastasis were determined using the Transwell assay. Scale bar = 500 μm; H: Cell apoptotic rates were detected after co-incubation using flow cytometry; I: Exosomes were isolated, and specific exosome markers were identified using western blotting; J: The particle size analyzer showed that the average diameter of the vesicles was within the diameter range of exosomes; K: The uptake of labeled exosomes was observed and verified; L-S: Hep3B cell viability and metastasis were both suppressed after exposure to mesenchymal stem cell (MSC)-derived exosomes, and apoptosis was enhanced; T: Comparison of miR-148a-3p and metal-regulatory transcription factor-1 (MTF-1) expression levels in MSC- and hepatocellular carcinoma (HCC)-derived exosomes; U: Assessment of miR-148a-3p and MTF-1 expression by real-time polymerase chain reaction in HCC cells co-cultured with MSC and MSC-derived exosomes. Quantitative data from three independent experiments are shown as the mean ± SD (error bars). a P < 0.05, b P < 0.01, c P < 0.001. HCC: Hepatocellular carcinoma; miRNA: MicroRNA; MSC: Mesenchymal stem cell; MTF-1: Metal-regulatory transcription factor-1; PBS: Phosphate buffered saline.

Article Snippet: The human HCC cell lines Hep3B (RRID: CVCL_0326, iCell Bioscience, Shanghai, China) and SNU387 (RRID: CVCL_0250, Procell, Wuhan, China), MSC (Procell, Wuhan, China), and the human embryonic kidney cell line HEK 293T (RRID: CVCL_0045, Cell Research, Shanghai, China) were cultured according to the manufacturer’s instructions.

Techniques: Derivative Assay, Incubation, Transwell Assay, Flow Cytometry, Isolation, Western Blot, Labeling, Comparison, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Saline

Journal: World Journal of Gastroenterology

Article Title: Exosome-mediated transfer of circRNA563 promoting hepatocellular carcinoma by targeting the microRNA148a-3p/metal-regulatory transcription factor-1 pathway

doi: 10.3748/wjg.v29.i46.6060

Figure Lengend Snippet: Identification of hsa_circRNA_0000563. A: Three circular RNAs (circRNAs) (circ562, circ563, and circ1110) were predicted to most likely bind to miR-148a-3p, and the miR-148a-3p and metal-regulatory transcription factor-1 (MTF-1) levels in hepatocellular carcinoma (HCC) cells transfected with the predicted circRNA were detected by quantitative real-time polymerase chain reaction (RT-PCR); B: A dual-luciferase reporter assay was performed to confirm the direct binding between circ563 and miR-148a-3p based on their complementary sequences; C: The knockdown efficiency was verified by RT-PCR; D: The expression levels of circ563 in exosomes were assessed by RT-PCR; E and F: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and colony-forming assay results showed that circ563 silencing suppressed cell proliferation and reduced the number of colonies; G: Knockdown of circ563 enhanced Hep3B cell apoptosis as determined by flow cytometry; H and I: Transwell assays revealed that reducing the levels of circ563 impaired the migratory potential and invasiveness of HCC cells. a P < 0.05, b P < 0.01, c P < 0.001. circRNA: Circular RNA; HCC: Hepatocellular carcinoma; Exo: Exosome.

Article Snippet: The human HCC cell lines Hep3B (RRID: CVCL_0326, iCell Bioscience, Shanghai, China) and SNU387 (RRID: CVCL_0250, Procell, Wuhan, China), MSC (Procell, Wuhan, China), and the human embryonic kidney cell line HEK 293T (RRID: CVCL_0045, Cell Research, Shanghai, China) were cultured according to the manufacturer’s instructions.

Techniques: Transfection, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Luciferase, Reporter Assay, Binding Assay, Knockdown, Expressing, Flow Cytometry

Journal: World Journal of Gastroenterology

Article Title: Exosome-mediated transfer of circRNA563 promoting hepatocellular carcinoma by targeting the microRNA148a-3p/metal-regulatory transcription factor-1 pathway

doi: 10.3748/wjg.v29.i46.6060

Figure Lengend Snippet: Exosomal circ563 facilitating hepatocellular carcinoma progression. A: First, a knockdown experiment was conducted in Hep3B cells, and the circ563 levels in Hep3B-derived exosomes were quantified by polymerase chain reaction. Hep3B cell function was assessed after co-treatment with exosomes isolated from the culture medium of circ563-knockdown Hep3B cells; B-D: Downregulation of exosomal circ563 (exo-circ563) reduced Hep3B cell proliferation and the number of colonies formed; E-H: Decreased exo-circ563 levels suppressed the migratory activity and invasiveness of the Hep3B cells; I: The downregulation of exo-circ563 was correlated with apoptosis induction; J: The efficiency of circ563 overexpression in the mesenchymal stem cells (MSCs) was determined by quantitative real-time polymerase chain reaction; K: The upregulation of circ563 in MSC-derived exosomes was confirmed; L-R: The hepatocellular carcinoma (HCC) cell proliferation, metastasis, and apoptosis rates were assessed following exosome treatment. Exo-circ563 significantly induced HCC cell proliferation, migration, and invasion; S: Flow cytometry analysis showed that the percentage of apoptotic cells was significantly decreased. a P < 0.05, b P < 0.01, c P < 0.001. exo-circ563: Exosomal circ563; HCC: Hepatocellular carcinoma; MSC: Mesenchymal stem cell; Exo: Exosome.

Article Snippet: The human HCC cell lines Hep3B (RRID: CVCL_0326, iCell Bioscience, Shanghai, China) and SNU387 (RRID: CVCL_0250, Procell, Wuhan, China), MSC (Procell, Wuhan, China), and the human embryonic kidney cell line HEK 293T (RRID: CVCL_0045, Cell Research, Shanghai, China) were cultured according to the manufacturer’s instructions.

Techniques: Knockdown, Derivative Assay, Polymerase Chain Reaction, Cell Function Assay, Isolation, Activity Assay, Over Expression, Real-time Polymerase Chain Reaction, Migration, Flow Cytometry

Journal: World Journal of Gastroenterology

Article Title: Exosome-mediated transfer of circRNA563 promoting hepatocellular carcinoma by targeting the microRNA148a-3p/metal-regulatory transcription factor-1 pathway

doi: 10.3748/wjg.v29.i46.6060

Figure Lengend Snippet: Oncogenic effect of exosomal circ563 prevented by either miR-148a-3p upregulation or metal-regulatory transcription factor-1 knockdown. A-P: Exosomal circ563 promoted Hep3B cell proliferation and invasion, which were reversed by miR-148a-3p overexpression or metal-regulatory transcription factor-1 depletion. Group 1: Co-culturing Hep3B cells with mesenchymal stem cell (MSC)-derived exosomes; Group 2: Co-culturing Hep3B cells with exosomes derived from circ563-overexpressing MSCs; Group 3: Co-culturing Hep3B cells after miR-148a-3p upregulation with exosomes derived from circ563-overexpressing MSCs; and Group 4: Co-culturing Hep3B cells after metal-regulatory transcription factor-1 knockdown with exosomes derived from circ563-overexpressing MSCs. a P < 0.05, b P < 0.01, c P < 0.001.

Article Snippet: The human HCC cell lines Hep3B (RRID: CVCL_0326, iCell Bioscience, Shanghai, China) and SNU387 (RRID: CVCL_0250, Procell, Wuhan, China), MSC (Procell, Wuhan, China), and the human embryonic kidney cell line HEK 293T (RRID: CVCL_0045, Cell Research, Shanghai, China) were cultured according to the manufacturer’s instructions.

Techniques: Knockdown, Over Expression, Derivative Assay

Journal: World Journal of Gastroenterology

Article Title: Exosome-mediated transfer of circRNA563 promoting hepatocellular carcinoma by targeting the microRNA148a-3p/metal-regulatory transcription factor-1 pathway

doi: 10.3748/wjg.v29.i46.6060

Figure Lengend Snippet: Circ563 promoting hepatocellular carcinoma tumor growth in vivo . Mouse xenografts were generated to verify the role of circ563 in vivo . Hepatocellular carcinoma cells stably transfected with miR-148a-3p overexpression, metal-regulatory transcription factor-1 knockdown, or control vectors were implanted into the subcutaneous tumors of mice. The xenografts were treated with exosomes after four weeks. A-C: Accelerated growth of circ563-overexpressing Hep3B-derived xenografts is shown with increased tumor volume and weight compared with the control group. Both miR-148a-3p overexpression and metal-regulatory transcription factor-1 (MTF-1) depletion partially reversed the increase in tumor volume and weight; D and E: MTF-1 staining reveals a similar trend with cell growth. a P < 0.05, b P < 0.01, c P < 0.001. exo-circ563: Exosomal circ563; HCC: Hepatocellular carcinoma; MSC: Mesenchymal stem cell; Exo: Exosome; MTF-1: Metal-regulatory transcription factor-1.

Article Snippet: The human HCC cell lines Hep3B (RRID: CVCL_0326, iCell Bioscience, Shanghai, China) and SNU387 (RRID: CVCL_0250, Procell, Wuhan, China), MSC (Procell, Wuhan, China), and the human embryonic kidney cell line HEK 293T (RRID: CVCL_0045, Cell Research, Shanghai, China) were cultured according to the manufacturer’s instructions.

Techniques: In Vivo, Generated, Stable Transfection, Transfection, Over Expression, Knockdown, Control, Derivative Assay, Staining

Journal: Biomimetics

Article Title: Anticancer Activity of Thiophene Carboxamide Derivatives as CA-4 Biomimetics: Synthesis, Biological Potency, 3D Spheroid Model, and Molecular Dynamics Simulation

doi: 10.3390/biomimetics7040247

Figure Lengend Snippet: IC 50 (µM) of phenyl-thiophene-carboxamide compounds ( 2a – 2e ) on several cell lines.

Article Snippet: Our results showed that the PSA of the most synthesized structures was biomimetic to CA-4, and similar chemical and biological properties were observed against Hep3B cancer cell line.

Techniques:

Journal: Biomimetics

Article Title: Anticancer Activity of Thiophene Carboxamide Derivatives as CA-4 Biomimetics: Synthesis, Biological Potency, 3D Spheroid Model, and Molecular Dynamics Simulation

doi: 10.3390/biomimetics7040247

Figure Lengend Snippet: RHA-3 ( 2b ) and RHA-6 ( 2e ) perturb 3D hepatocellular spheroids’ formation. Images of cluster/s formed by Hep3B hepatocellular carcinoma in presence of 2b (17 µg/mL) or 2e (17 µg/mL) after 24 h of treatment compared to controls ( A ). Cluster percentage of occupied area relative to the negative control ( B ), cluster circularity ( C ), and cluster count ( D ). The non-treated cells are referred to as a negative control. At 100 μg/mL, DOX was utilized as a positive control. The scale bar represents a distance of 10 μm. Circularity scale: a value of 1 represents a perfect circle (ns: p > 0.05, **: p ≤ 0.01, ***: p ≤ 0.001, and ****: p ≤ 0.0001).

Article Snippet: Our results showed that the PSA of the most synthesized structures was biomimetic to CA-4, and similar chemical and biological properties were observed against Hep3B cancer cell line.

Techniques: Negative Control, Positive Control