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: British Journal of Cancer

Article Title: Inhibiting CBX4 efficiently protects hepatocellular carcinoma cells against sorafenib resistance

doi: 10.1038/s41416-020-01240-6

Figure Lengend Snippet: a The diagram illustrates how miR424 was identified from the miRNA sequencing data and predicted by Venn screening from miRNA database websites. b Sequence alignment of the human miR424 seed sequence with the 3’-UTR of CBX4. The mutated sequence in the matched binding sites for the gene that was used to create the firefly luciferase reporter constructs is shown at the bottom of the gene set. A luciferase reporter assay demonstrated that miR424 inhibited the transcription of the wild-type but not the mutant 3’-UTRs of CBX4. c The expression of endogenous CBX4 was inhibited in miR424-overexpressed Huh7-SR and PLC-SR cells. d In contrast, CBX4 levels were increased in Huh7 and PLC cells with miR424-TUD. All data were compared with the respective controls, and the mRNA level was detected by qRT-PCR. CBX4 mRNA expression was normalised to that of GAPDH mRNA; and three independent experiments were conducted. e Relative miR424 expression in HCC tissues and matched adjacent normal tissues as assessed by qRT-PCR. f , g Relative expression data of miR424 in HCC cases were further analysed. The negative relationship between miR424 expression and liver cirrhosis ( f ) and size ( g ). h , i Kaplan–Meier curves of disease-free survival (DFS) ( h ) and overall survival (OS) ( i ). Survival of the high and low miR424 expression groups assessed using log-rank (Mantel–Cox) test in HCC, which were divided according to a cut-off of 2.5, the median value of CBX4 mRNA expression relative to GAPDH mRNA. j From the TCGA database, tumour purity was highly negatively correlated with miR424 expression. k Linear regression and correlation between the miR424 and CBX4 mRNA levels in 341 HCC tissues from the TCGA database. l The negative linear regression and correlation analysis for the relation of the mRNA levels of CBX4 and those of miR424 in 106 HCC patients by qRT-PCR.

Article Snippet: The human HCC cell lines PLC (catalogue number: CRL-8024TM) was obtained from the Global Bioresource Center of American Type Culture Collection (ATCC) and Huh7 (catalogue number: JCRB0403) was purchased from Health Science Research Resources Bank (Osaka, Japan).

Techniques: Sequencing, Binding Assay, Luciferase, Construct, Reporter Assay, Mutagenesis, Expressing, Quantitative RT-PCR

Journal: Journal of Hepatocellular Carcinoma

Article Title: Comparative Response of HCC Cells to TKIs: Modified in vitro Testing and Descriptive Expression Analysis

doi: 10.2147/JHC.S356333

Figure Lengend Snippet: Clinical and Histological Features of Nine Hepatoma Cell Lines

Article Snippet: HuH7 , Asian , M , 57 , (-) , (+) , HCC , RIKEN, JCRB , [ ] .

Techniques:

Journal: PLoS ONE

Article Title: Soluble factors from adipose tissue-derived mesenchymal stem cells promote canine hepatocellular carcinoma cell proliferation and invasion

doi: 10.1371/journal.pone.0191539

Figure Lengend Snippet: (A, B) Representative images of Transwell assays observed under a light microscope in the cultures without (A) and (B) with AT-MSC-CM. (C) The percent of invading HCC cells was significantly increased in cells cultured with AT-MSC-CM compared with control cells cultured without AT-MSC-CM. Data are expressed as mean ± SD. ** P < 0.01.

Article Snippet: AZACH (Cosmo Bio, Tokyo, Japan), a canine HCC cell line, was cultured in H-DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution in a humidified atmosphere of 5% CO 2 at 37°C.

Techniques: Light Microscopy, Cell Culture, Control

Journal: PLoS ONE

Article Title: Soluble factors from adipose tissue-derived mesenchymal stem cells promote canine hepatocellular carcinoma cell proliferation and invasion

doi: 10.1371/journal.pone.0191539

Figure Lengend Snippet: Expression of mRNA of the indicated growth factors and MMPs were evaluated in canine HCC cells cultured with AT-MSC-CM compared with control HCC cells cultured without AT-MSC-CM. Data are expressed as mean ± SD. * P < 0.05; ** P < 0.01.

Article Snippet: AZACH (Cosmo Bio, Tokyo, Japan), a canine HCC cell line, was cultured in H-DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution in a humidified atmosphere of 5% CO 2 at 37°C.

Techniques: Expressing, Cell Culture, Control