normal liver epithelial cells thle 3  (ATCC)

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Nucleolar factors associated with FC/DFC are upregulated at both mRNA and protein level in HCC. Relative mRNA levels of ( A ) TCOF1 (Treacle), ( B ) UBF, ( C ) FBL (Fibrillarin), ( D ) NCL (nucleolin), and ( E ) NPM1 (nucleophosmin) measured by qPCR in HCC cells (PLC/PRF/5 and SNU-449) and control cells (THLE-3). GAPDH was used as the housekeeping gene, and fold change was determined relative to mRNA levels in THLE-3 cells. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD, with statistical significance assessed by one-way ANOVA and indicated with numerical P -values. Protein levels of ( F ) Treacle, ( G ) UBF, ( H ) Fibrillarin, ( I ) nucleolin, and ( J ) nucleophosmin measured by WB in HCC cells (PLC/PRF/5 and SNU-449) and control cells (THLE-3). α-tubulin was used as a loading control (LC). The molecular weight in kDa is listed on the left side of the blots (left panel), and protein signals were determined in each cell line relative to the loading control (right panel). Individual points in the graphs represent biological replicates ( n = 3). The graph depicts the mean ± SD, with statistical significance assessed by one-way ANOVA and indicated by numerical P -values.

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Control, Molecular Weight

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Nucleolar expression of FC/DFC-associated factors is upregulated in HCC. Representative images of ( A ) Treacle (green), ( B ) UBF (green), ( C ) Fibrillarin (green), and ( D ) nucleolin (green) and nucleophosmin (magenta), and Pol II (yellow) in THLE-3, PLC/PRF/5, and SNU-449 cells. ( E ) Total nucleolar Treacle intensity per nucleus. Graphs depict one representative replicate ( n = 3) with data points representing individual cells and the median indicated by the red line. A Kruskal–Wallis test was applied to assess differences between groups for all three biological replicates combined, and statistical significance is indicated by numerical P -values. ( F ) Total nucleolar UBF intensity per nucleus otherwise as in panel (E). ( G ) Total nucleolar Fibrillarin intensity per nucleus, otherwise as in panel (E). ( H ) Total nucleolar nucleolin intensity per nucleus otherwise as in panel (E). (I) Total nucleolar nucleophosmin intensity per nucleus otherwise as in panel (E).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Expressing

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: rDNA transcription is increased in HCC. ( A ) Representative images of EU incorporation (green). THLE-3, PLC/PRF/5, and SNU-449 cells were stained with DAPI (blue) and an antibody against Pol II (yellow). ( B ) Total nucleolar EU intensity per nucleus as a readout for rDNA transcription in THLE-3, PLC/PRF/5, and SNU-449 cells. The graph depicts one representative biological replicate ( n = 3), with data points representing individual cells, and the median indicated by the red line. A Kruskal–Wallis test was applied to assess differences between groups for the replicates combined, and statistical significance is indicated by numerical P -values. ( C ) Nucleolar size measured through an inverse intensity-based mask of Pol II in THLE-3, PLC/PRF/5, and SNU-449 cells. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD, and statistical significance assessed by one-way ANOVA is indicated with numerical P -values. ( D ) Number of nucleoli per nucleus in THLE-3, PLC/PRF/5, and SNU-449 cells measured through an inverse intensity-based mask of Pol II. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD, and statistical significance assessed by one-way ANOVA is indicated with numerical P -values. ( E ) Total nucleolar area otherwise as in panel (C).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Staining

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Anti-cancer drugs targeting the nucleolus inhibit cell viability in HCC. Drug response curves with corresponding IC 50 values and 95% CIs. Drugs were applied for 72 h, after which cell viability was assessed in THLE-3, PLC/PRF/5, and SNU-449 cells. ( A ) Drug response curve for CX-5461 with obtained IC 50 values and 95% CI (left panel). The graph shows the obtained IC 50 values represented as the mean ± SD (right panel). Individual points in the graph represent biological replicates ( n = 3). Statistical significance was assessed by one-way ANOVA and are indicated with numerical P -values. ( B ) Drug response curve for BMH-21 otherwise as in panel (A). ( C ) Drug response curve for Oxaliplatin, otherwise as in panel (A). ( D ) Drug response curve for JP-1302 otherwise as in panel (A). ( E ) Drug response curve for Sorafenib otherwise as in panel (A).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques:

Journal: NAR Cancer

Article Title: Ribosome biogenesis is increased in hepatocellular carcinoma and represents a potential therapeutic target

doi: 10.1093/narcan/zcaf058

Figure Lengend Snippet: Nucleolar-targeting compounds inhibit nucleolar activity and induce selective DNA damage. ( A ) Representative images of EU incorporation (green) in THLE-3, PLC/PRF/5, and SNU-449 cells 24 h after treatment with four anti-cancer drugs targeting the nucleolus: CX-5461, BMH-21, Oxaliplatin, and JP-1302, or targeted therapy Sorafenib. Cells were treated with DMSO as control and were stained with DAPI (blue) and Pol II (yellow). ( B ) Mean nucleolar EU intensity per nucleus as a readout for rDNA transcription in THLE-3, PLC/PRF/5, and SNU-449 cells 24 h after treatment with four anti-cancer drugs targeting the nucleolus: CX-5461, BMH-21, Oxaliplatin, and JP-1302, or targeted therapy Sorafenib. Cells were treated with DMSO as control. The graph depicts one representative biological replicate ( n = 3), with data points representing individual cells, and the median indicated by the red line. A Kruskal–Wallis test was applied to assess differences between groups for the replicates combined, and statistical significance is indicated by numerical P -values. ( C ) Representative images of nuclear 53BP1 and γH2AX foci accumulation in THLE-3, PLC/PRF/5, and SNU-449 cells 24 h after treatment with four anti-cancer drugs targeting the nucleolus: CX-5461, BMH-21, Oxaliplatin, and JP-1302, or targeted therapy Sorafenib. Cells were treated with DMSO as control, and stained with DAPI (blue), 53BP1 (yellow), and γH2AX (red). ( D ) Fold change of nuclear 53BP1 foci per cell relative to DMSO. Individual points in the graph represent biological replicates ( n = 3). The graph depicts the mean ± SD and statistical significance was assessed by one sample t test and indicated by numerical P -values. ( E ) Fold change of nuclear γH2AX foci per cell relative to DMSO otherwise as in panel (D).

Article Snippet: Human osteosarcoma U2OS cells (#HTB-96), HCC cell lines PLC/PRF/5 (#CRL-8024), SNU-449 (#CRL-2234), and normal liver epithelial cells THLE-3 (#CRL-3583) were purchased from the American Type Culture Collection.

Techniques: Activity Assay, Control, Staining

Journal: BMC Cancer

Article Title: Circ_0064288 acts as an oncogene of hepatocellular carcinoma cells by inhibiting miR-335-5p expression and promoting ROCK1 expression

doi: 10.1186/s12885-022-09323-8

Figure Lengend Snippet: Circ_0064288 expression is up-modulated in HCC. A - B . Volcano plots were utilized to show the differentially expressed circRNAs in HCC, with the data of GSE94508 and GSE97332. C . Venn diagram was employed to screen the circRNAs that were remarkably up-modulated in both datasets. D . Circ_0064288 expression in HCC tissues and paracancerous tissues was examined by qRT-PCR, and then the fold change of circ_0064288 expression in each HCC sample was shown. E . Kaplan-Meier curve was applied to analyze the relationship between circ_0064288 expression and the overall survival of HCC patients. F . Circ_0064288 expression in normal hepatic epithelial cell line (THLE-3) and HCC cell lines (Huh7, Hep3B, HCCLM3, and MHCC97-L) was measured by qRT-PCR. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human HCC cell lines (Huh7, Hep3B, HCCLM3, and MHCC97-L) and human normal liver epithelial cell line (THLE-3) were procured from American Type Culture Collection (ATCC, Manassas, VA, USA) and Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: BMC Cancer

Article Title: Circ_0064288 acts as an oncogene of hepatocellular carcinoma cells by inhibiting miR-335-5p expression and promoting ROCK1 expression

doi: 10.1186/s12885-022-09323-8

Figure Lengend Snippet: Circ_0064288 functions as ceRNA and sponges miR-335-5p. A . Bioinformatics databases (StarBase and Circular RNA Interactome) were used to predict the binding site of circ_0064288 to miR-335-5p. B . qRT-PCR was employed to detect circ_0064288 expression in the cytoplasm and nucleus of Huh7 and Hep3B cells. C . Dual-luciferase reporter gene experiment showed that the predicted binding sequence between circ_0064288 and miR-335-5p was functional. D . qRT-PCR was employed to detect miR-335-5p expression in HCC tissues and paracancerous tissues. E . qRT-PCR was used to detect miR-335-5p expression in normal hepatic epithelial cell line (THLE-3) and HCC cell lines (Huh7, Hep3B, HCCLM3, and MHCC97-L). F . The correlation between circ_0064288 expression and miR-335-5p expression in HCC tissues was evaluated by Pearson’s correlation coefficient. *** P < 0.001, ns, means no statistical significance. *** P < 0.001, ns, means no statistical significance

Article Snippet: Human HCC cell lines (Huh7, Hep3B, HCCLM3, and MHCC97-L) and human normal liver epithelial cell line (THLE-3) were procured from American Type Culture Collection (ATCC, Manassas, VA, USA) and Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China).

Techniques: Binding Assay, Quantitative RT-PCR, Expressing, Luciferase, Sequencing, Functional Assay