Journal: International Journal of Molecular Sciences

Article Title: ER Stress-Activated HSF1 Governs Cancer Cell Resistance to USP7 Inhibitor-Based Chemotherapy through the PERK Pathway

doi: 10.3390/ijms25052768

Figure Lengend Snippet: HSR and UPR gene expression in USP7 inhibitor (USP7i)-resistant cancer cells. ( A , B ) Cell viability analysis in parental (P) and USP7i-resistant (R) HeLa and H1299 cells in the absence or presence of P22077 ( A ) or P5091 ( B ) for 24 h. Values represent the mean ± SD (n = 4). * p < 0.05, ** p < 0.0, and *** p < 0.001 by one-way ANOVA. Tukey’s post hoc test was performed for statistical analysis. ( C ) Venn diagram showing the 19 common genes upregulated in parental vs P22077-resistant HeLa or H1299 cells. ( D ) Summary of the main GO terms obtained for biological process (BP) in the comparisons of parental vs P22077-resistant HeLa and H1299 cells. ( E ) Smear plot of the log2 fold change (FC) gene expression of P22077-resistant cells compared to parental cells. Blue points represent all upregulated or downregulated genes. Red points represent USP7i-induced HSR- and UPR-associated genes.

Article Snippet: HeLa (10002), H1299 (25803), HEK293 (21573), and Hep3B (88064) cancer cells were obtained from Korean Cell Line Bank (Seoul, Republic of Korea).

Techniques: Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: ER Stress-Activated HSF1 Governs Cancer Cell Resistance to USP7 Inhibitor-Based Chemotherapy through the PERK Pathway

doi: 10.3390/ijms25052768

Figure Lengend Snippet: HSR and UPR gene expression in response to USP7i. ( A , B ) HSR- and UPR-related gene expression in P22077 (10 µM)-treated HeLa ( A ), HEK293 ( B , upper), and H1299 ( B , bottom) cells in a time-dependent manner. ( C ) HSR and UPR gene expression in P5091 (10 µM)-treated HeLa (upper) and H1299 (bottom) cells. The values represent the mean ± SD (n = 3); * p < 0.05, ** p < 0.01, and *** p < 0.001. One-way ANOVA Tukey’s post hoc test was performed for statistical analysis.

Article Snippet: HeLa (10002), H1299 (25803), HEK293 (21573), and Hep3B (88064) cancer cells were obtained from Korean Cell Line Bank (Seoul, Republic of Korea).

Techniques: Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: ER Stress-Activated HSF1 Governs Cancer Cell Resistance to USP7 Inhibitor-Based Chemotherapy through the PERK Pathway

doi: 10.3390/ijms25052768

Figure Lengend Snippet: HSF1 phosphorylation by USP7i. ( A , B ) A shift in the molecular weight of HSF1 in P22077 ( A ) or P5091 ( B )-treated cells. H1299, Hep3B, HEK293. and HeLa cells were incubated with 10 µM of P22077 or P5091 for 1 h or 2 h. ( C ) The effect of lambda protein phosphatase (pptase) in USP7i induced a shift in the molecular weight of HSF1. HeLa cells were incubated with 10 µM of P22077 for 2 h, and then total cell lysates were incubated in the absence or presence of pptase for 1 h at 37 °C. ( D ) HEK293 cells were transiently transfected with full-length Flag-HSF1, and cells were then incubated with DMSO or 10 µM of P22077 for 2 h. Phosphorylated HSF1 was measured using immunoprecipitation and Western blotting with indicated antibodies. ( E ) HeLa cells were incubated with inhibitors targeting ubiquitin-activating enzyme E1 (PYR41, 10 µM), sumoylation (TAK981, 1 µM), and glycosylation (NGI1, 10 µM) for 1 h, and the cells were then further incubated in the absence or presence of 10 µM of P22077 for 2 h. ( F ) A shift in the molecular weight of HSF1 in parental or P22077-resistant cells. Parental cells were incubated with 10 µM of P22055 or 10 µM of P5091 for 1 h. ( G ) Occupancy of HSF1 on the promoter region of HSPA1A , HSPA1B, and HSPA6 in parental (HeLa-P) or P22077-resistant (HeLa-R) cells.

Article Snippet: HeLa (10002), H1299 (25803), HEK293 (21573), and Hep3B (88064) cancer cells were obtained from Korean Cell Line Bank (Seoul, Republic of Korea).

Techniques: Phospho-proteomics, Molecular Weight, Incubation, Transfection, Immunoprecipitation, Western Blot, Ubiquitin Proteomics, Glycoproteomics

Journal: International Journal of Molecular Sciences

Article Title: ER Stress-Activated HSF1 Governs Cancer Cell Resistance to USP7 Inhibitor-Based Chemotherapy through the PERK Pathway

doi: 10.3390/ijms25052768

Figure Lengend Snippet: Suppression of HSF1 and PERK sensitizes USP7i-induced cytotoxicity. ( A , B ) Cell viability in control (shCtrl) or HSF1 (shHSF1)-silenced parental (P) and USP7i-resistant (R) HeLa ( A ) and H1299 ( B ) cells. Cells were incubated with P22077 and P5091 for 24 h in a dose-dependent manner (2, 5, 10, 20, and 40 µM) as indicated. ( C ) Parental and USP7i-resistant HeLa cells were pre-incubated with KRIBB11 (2 µM) for 2 h, and the cells were then further incubated with P22077 (10 µM) for 24 h. ( D ) Cell viability in control or PERK silenced parental and USP7i-resistant HeLa cells. ( E ) Parental and USP7i-resistant HeLa cells were pre-incubated with PERKi (100 nM) for 2 h, and cells were then further incubated with P22077 (10 µM) for 24 h. Values represent the mean ± SD (n = 4); * p < 0.05 and ** p < 0.01. One-way ANOVA Tukey’s post hoc test was performed for statistical analysis.

Article Snippet: HeLa (10002), H1299 (25803), HEK293 (21573), and Hep3B (88064) cancer cells were obtained from Korean Cell Line Bank (Seoul, Republic of Korea).

Techniques: Control, Incubation

Journal: International Journal of Oncology

Article Title: Anti-angiogenic effect of Bryopsis plumosa -derived peptide via aquaporin 3 in non-small cell lung cancer

doi: 10.3892/ijo.2024.5711

Figure Lengend Snippet: Expression of AQP3 in lung cancer. (A) Levels of AQP3 in lung cancer (H1299) and normal cells (MRC5 and HUVEC). (B) Gene expression analysis of AQP3 in normal and cancer lung tissue using Gene Expression Profiling Interactive Analysis. * P<0.05 (C) Kaplan-Meier survival graph of patients with lung cancer according to AQP3 expression. AQP, aquaporin; HUVEC, human umbilical vein endothelial cell.

Article Snippet: The human H1299 lung cancer cell and MRC5 lung normal fibroblast cell lines (Korean Cell Line Bank; cat. no. 25803 and 10171) were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and 1% antibiotics.

Techniques: Expressing, Gene Expression