Journal: Cell Stress & Chaperones

Article Title: Heat stress-induced heat shock protein 90 alpha family class A member 1 upregulation stabilizes yes-associated protein through ubiquitination inhibition to boost hepatic cancer radiofrequency hyperthermia resistance

doi: 10.1016/j.cstres.2025.100140

Figure Lengend Snippet: HSP90AA1 plays a pivotal role in the heat stress response of liver cancer cells. (a) WB analysis was employed to assess alterations in the expression levels of HSP110, HSP90AA1, HSP70, and HSP60 in liver cancer cells following exposure to heat stress. (b) To evaluate transfection efficiency, WB analysis was performed to assess the effects of HSP110 or HSP90AA1 silence in HepG2 and Huh7 cells. (c) MTT assay was used to evaluate changes in cell viability of HepG2 and Huh7 cells after heat stress treatment with or without the silencing of HSP90 and HSP110, respectively. (d) Flow cytometry was used to analyze changes in the apoptosis rate of HepG2 and Huh7 cells after heat stress treatment with or without the silencing of HSP90 and HSP110, respectively. All experiments were performed with at least three replicates. Statistical significance was denoted as * P < 0.05 and ** P < 0.01.

Article Snippet: HSP90 , Proteintech , 13171-1-AP , 1:2000.

Techniques: Expressing, Transfection, MTT Assay, Flow Cytometry

Journal: Cell Stress & Chaperones

Article Title: Heat stress-induced heat shock protein 90 alpha family class A member 1 upregulation stabilizes yes-associated protein through ubiquitination inhibition to boost hepatic cancer radiofrequency hyperthermia resistance

doi: 10.1016/j.cstres.2025.100140

Figure Lengend Snippet: Bioinformatics analysis of the relationship between HSP90AA1 and its related DEGs and the clinical relevance of HSP90AA1 in HCC. (a) According to the average expression level of HSP90AA1AA1 in HCC, the samples were divided into the HSP90AA1AA1 high expression group and the low expression group, with the sample data downloaded from the TCGA-LIHC dataset. HSP90AA1AA1-related DEGs were shown by a volcano plot (right panel) and a heat map (left panel). (b) The KEGG analysis was utilized to determine the biological processes and signaling pathways in which the DEGs were predominantly involved. (c) The intersecting genes between the genes in the Melotome database and the DEGs were utilized to construct a Venn diagram. (d) KEGG enrichment analysis was employed to elucidate the primary biological processes and signaling pathways implicated in the intersectional genes. (e) The GEPIA database was used to assess HSP90AA1AA1-OS survival curves in HCC. (f) The UALCAN database analyzed the expression of HSP90AA1AA1 in normal and tumor. (g) The UALCAN database evaluated HSP90AA1AA1 expression across HCC stages.

Article Snippet: HSP90 , Proteintech , 13171-1-AP , 1:2000.

Techniques: Expressing, Protein-Protein interactions, Construct

Journal: Cell Stress & Chaperones

Article Title: Heat stress-induced heat shock protein 90 alpha family class A member 1 upregulation stabilizes yes-associated protein through ubiquitination inhibition to boost hepatic cancer radiofrequency hyperthermia resistance

doi: 10.1016/j.cstres.2025.100140

Figure Lengend Snippet: The activation of the YAP pathway under heat stress conditions confers a protective effect in liver cancer cells. (a) WB analysis was utilized to evaluate protein changes in the Hippo-YAP pathway in HepG2 and Huh7 cells following exposure to heat stress. (b) The MTT assay was utilized to evaluate variations in cell viability in HepG2 and Huh7 cells following heat stress with or without TDI-011536 intervention. (c) Flow cytometry was conducted to analyze changes in apoptosis rates in HepG2 and Huh7 cells after heat stress treatment with or without TDI-011536. (d) WB analysis was employed to assess alterations in the levels of p-YAP, total YAP protein expression, and the p-YAP/YAP ratio in HepG2 and Huh7 cells after exposure to heat stress with or without TDI-011536 treatment. (e) WB analysis was performed to evaluate the changes in p-YAP levels, total YAP protein expression, TAZ protein levels, and the p-YAP/YAP ratio in liver cancer cells with HSP90AA1 silence under heat stress conditions. All experiments were performed with at least three replicates. Statistical significance was denoted as * P < 0.05 and ** P < 0.01.

Article Snippet: HSP90 , Proteintech , 13171-1-AP , 1:2000.

Techniques: Activation Assay, MTT Assay, Flow Cytometry, Expressing

Journal: Cell Stress & Chaperones

Article Title: Heat stress-induced heat shock protein 90 alpha family class A member 1 upregulation stabilizes yes-associated protein through ubiquitination inhibition to boost hepatic cancer radiofrequency hyperthermia resistance

doi: 10.1016/j.cstres.2025.100140

Figure Lengend Snippet: HSP90AA1 regulates HT response in HCC by modulating YAP phosphorylation and activity. (a-c) These panels validate the efficiency of HSP90AA1 and YAP silencing and overexpression (YAP-OE, siR-YAP, and HSP90AA1-OE) in HepG2 cells using WB. The effects of co-treating HepG2 cells with siR-HSP90AA1 + YAP-OE or HSP90AA1-OE + siR-YAP under HT were assessed by WB analysis of YAP and p-YAP (d, e), quantification of apoptosis rates using flow cytometry (f, g), and measurement of cell viability via MTT assay (h, i). All experiments were performed with at least three replicates. Statistical significance was denoted as * P < 0.05 and ** P < 0.01.

Article Snippet: HSP90 , Proteintech , 13171-1-AP , 1:2000.

Techniques: Phospho-proteomics, Activity Assay, Over Expression, Flow Cytometry, MTT Assay

Journal: Cell Stress & Chaperones

Article Title: Heat stress-induced heat shock protein 90 alpha family class A member 1 upregulation stabilizes yes-associated protein through ubiquitination inhibition to boost hepatic cancer radiofrequency hyperthermia resistance

doi: 10.1016/j.cstres.2025.100140

Figure Lengend Snippet: HSP90AA1 promotes YAP protein stability by inhibiting its ubiquitination. (a) WB analysis was performed to detect changes in protein levels within the YAP signaling pathway in HepG2 and Huh7 cells after LATS1 silence under heat stress treatment or its absence. (b) Molecular docking analysis identified potential binding sites between HSP90AA1AA1 (in red) and YAP1 (in green) proteins, which were visualized using PyMOL software. (c) A Co-IP experiment was utilized to examine the relationship between HSP90AA1 and YAP in HepG2 cells treated with heat stress. (d) In HepG2 cells, HSP90AA1 was overexpressed, and WB analysis was used to assess changes in YAP protein levels following treatment with Cycloheximide (CHX, MCE, NJ, USA, HY-12320) at different time points. (e) A Co-IP experiment was employed to compare the levels of YAP ubiquitination between the proteasome inhibitor treatment group (MG132 group) and the group treated with both HSP90AA1 overexpression and MG132 in HepG2 cells. All experiments were performed with at least three replicates. Statistical significance was denoted as * P < 0.05 and ** P < 0.01.

Article Snippet: HSP90 , Proteintech , 13171-1-AP , 1:2000.

Techniques: Ubiquitin Proteomics, Binding Assay, Software, Co-Immunoprecipitation Assay, Over Expression

Journal: Cell Stress & Chaperones

Article Title: Heat stress-induced heat shock protein 90 alpha family class A member 1 upregulation stabilizes yes-associated protein through ubiquitination inhibition to boost hepatic cancer radiofrequency hyperthermia resistance

doi: 10.1016/j.cstres.2025.100140

Figure Lengend Snippet: Silencing HSP90AA1 enhances the sensitivity of the liver cancer xenograft model to RFH in vivo. Three groups were treated as follows: 1) sh-NC, serving as a control group; 2) sh-NC+RFH, where mice received RFH treatment; and 3) sh-HSP90AA1+RFH, where mice underwent RFH treatment after inoculation with liver cancer cells transfected with stable silencing of HSP90AA1. (a) In the construction of stable silencing HSP90AA1 in HepG2 cells, WB analysis was employed to assess the transfection efficiency ( n = 3). (b) Tumor imaging from mice, along with measurements of tumor weight and volume ( n = 6). (c) The changes of YAP and Ki67 proteins in tumor tissues were evaluated using IHC ( n = 3). (d) WB was employed to detect the levels of YAP, p-YAP proteins, and the p-YAP/YAP ratio in tumor tissues ( n = 3). Statistical significance was denoted as * P < 0.05 and ** P < 0.01.

Article Snippet: HSP90 , Proteintech , 13171-1-AP , 1:2000.

Techniques: In Vivo, Control, Transfection, Imaging