a2780 sk ov  (ATCC)

Journal: Communications Biology

Article Title: STUB1 suppresses paclitaxel resistance in ovarian cancer through mediating HOXB3 ubiquitination to inhibit PARK7 expression

doi: 10.1038/s42003-024-07127-z

Figure Lengend Snippet: a STUB1 expression was measured in tumor tissues and adjacent tissues from OC patients using RT-qPCR. b STUB1 expression was examined in chemotherapy resistant and chemotherapy sensitive samples from OC patients using RT-qPCR. c IC 50 value of PTX in SKOV3/R and A2780/R cells was detected by CCK-8. d STUB1 expression was examined in PTX-resistant or sensitive OC cells by RT-qPCR. * P < 0.05; ** P < 0.01.

Article Snippet: SKOV3 and A2780 cell lines were purchased from ATCC (USA).

Techniques: Expressing, Quantitative RT-PCR, CCK-8 Assay

Journal: Communications Biology

Article Title: STUB1 suppresses paclitaxel resistance in ovarian cancer through mediating HOXB3 ubiquitination to inhibit PARK7 expression

doi: 10.1038/s42003-024-07127-z

Figure Lengend Snippet: a , b STUB1 expression was examined in oe-STUB1-transfected PTX-resistant OC cells by RT-qPCR and western blot. SKOV3/R and A2780/R cells were transfected with oe-STUB1 upon PTX treatment. c Cells viability was examined using CCK-8. d Cells proliferation was evaluated using clone formation assay. SKOV3/R and A2780/R cells were transfected with oe-STUB1 upon PTX and Fer-1 treatments. e Mitochondrial morphology was monitored using a TEM. ( f ) Cell viability was detected using CCK-8. g – j ROS, iron, MDA, and GSH were detected using suitable kits. k PARK7 and GPX4 levels were determined using western blot. N = 3. * P < 0.05; ** P < 0.01, *** P < 0.001.

Article Snippet: SKOV3 and A2780 cell lines were purchased from ATCC (USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Tube Formation Assay

Journal: Communications Biology

Article Title: STUB1 suppresses paclitaxel resistance in ovarian cancer through mediating HOXB3 ubiquitination to inhibit PARK7 expression

doi: 10.1038/s42003-024-07127-z

Figure Lengend Snippet: a STUB1 and HOXB3 levels were determined in oe-STUB1-transfected SKOV3/R and A2780/R cells using western blot. b The interaction between STUB1 and HOXB3 was validated by Co-IP. c HOXB3 level was determined in oe-STUB1-transfected PTX-resistant OC cells upon MG132 treatment using western blot. d The degradation of HOXB3 protein was detected in oe-STUB1-transfected SKOV3/R and A2780/R cells upon CHX treatment using western blot. e HOXB3 ubiquitination was examined in oe-STUB1-transfected PTX-resistant OC cells using western blot. N = 3. * P < 0.05; ** P < 0.01, *** P < 0.001.

Article Snippet: SKOV3 and A2780 cell lines were purchased from ATCC (USA).

Techniques: Transfection, Western Blot, Co-Immunoprecipitation Assay, Ubiquitin Proteomics

Journal: Communications Biology

Article Title: STUB1 suppresses paclitaxel resistance in ovarian cancer through mediating HOXB3 ubiquitination to inhibit PARK7 expression

doi: 10.1038/s42003-024-07127-z

Figure Lengend Snippet: a , b HOXB3 expression was examined in sh-HOXB3-transfected SKOV3/R and A2780/R cells by RT-qPCR and western blot. sh-STUB1 with/without sh-HOXB3 was transfected into SKOV3/R and A2780/R cells upon PTX treatment. c Cells viability was examined using CCK-8. d Cells proliferation was evaluated using clone formation assay. e Mitochondrial morphology was monitored using a TEM. f – i ROS, iron, MDA and GSH were detected using suitable kits. j PARK7 and GPX4 levels were determined using western blot. N = 3. * P < 0.05; ** P < 0.01, *** P < 0.001.

Article Snippet: SKOV3 and A2780 cell lines were purchased from ATCC (USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Tube Formation Assay

Journal: Communications Biology

Article Title: STUB1 suppresses paclitaxel resistance in ovarian cancer through mediating HOXB3 ubiquitination to inhibit PARK7 expression

doi: 10.1038/s42003-024-07127-z

Figure Lengend Snippet: a , b PARK7 expression was examined in sh-PARK7-transfected SKOV3/R and A2780/R cells by RT-qPCR and western blot. sh-PARK7 with/without oe-HOXB3 or sh-STUB1 was transfected into PTX-resistant OC cells upon PTX treatment. c Cells viability was examined using CCK-8. d Cells proliferation was evaluated using clone formation assay. e Mitochondrial morphology was monitored using a TEM. f – i ROS, iron, MDA, and GSH were detected using suitable kits. j PARK7 and GPX4 levels were determined using western blot. N = 3. * P < 0.05; ** P < 0.01, *** P < 0.001.

Article Snippet: SKOV3 and A2780 cell lines were purchased from ATCC (USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Tube Formation Assay

Journal: Cancer Medicine

Article Title: BIRC5 facilitates cisplatin‐chemoresistance in a m 6 A ‐dependent manner in ovarian cancer

doi: 10.1002/cam4.6811

Figure Lengend Snippet: Establishment of the cisplatin‐resistant cells and identification of their gene expression profile in OC. (A) SKOV3/DDP, A2780/DDP, SKOV3 and A2780 treated with different doses of cisplatin for 24 h, followed by CCK8 assay. Dose‐response curve of cisplatin treatment in multiple OC line. (B) IC 50 were measured after treating increasing dose of cisplatin for 24 h (log10 scaled). (C) Colony formation treated with cisplatin 20 μM for 2 weeks. (D,E) Heatmap and volcano plot represent the differentially expressed mRNA in cisplatin resistant SKOV3/DDP and counterpart SKOV3 cells. The red and green represent upregulated and down‐regulated mRNA respectively. The arrow indicates BIRC5. The mean ± SD of triplicate experiments was plotted ( n = 3), * p < 0.05.

Article Snippet: Ovarian carcinoma cell lines SKOV3 and A2780 were obtained from American Type Culture Collection (ATCC).

Techniques: Gene Expression, CCK-8 Assay

Journal: Cancer Medicine

Article Title: BIRC5 facilitates cisplatin‐chemoresistance in a m 6 A ‐dependent manner in ovarian cancer

doi: 10.1002/cam4.6811

Figure Lengend Snippet: Down‐regulating BIRC5 reverses chemotherapy resistance of OC cells. (A) Relative expression of BIRC5 in OC tissues ( n = 30) and normal ovarian tissues ( n = 30) was detected by RT‐qPCR ( p < 0.05). (B) The abundance of BIRC5 transcripts from the TCGA database and GTEX database. (C) Expression of BIRC5 in cisplatin resistance cell and parental cells was detected by RT‐qPCR and Western blotting. (D) SKOV3/DDP and A2780/DDP were transfected with control shRNA and BIRC5 shRNA, and the effect of knockdown BIRC5 measured by Western blotting and RT‐qPCR. (E,F) Then the cell viability monitored by CCK8, and IC 50 after treating increasing dose of cisplatin for 24 h. (G,H) EdU and colony formation assay were used to assess cell survival of knocked‐down BIRC5 cisplatin resistance cells after 20 μM cisplatin treatment. (I) Cell apoptosis was measured by flow cytometry in SKOV3/DDP and A2780/DDP with indicated treatment, and right histograms represent apoptosis rate (early + late). The mean ± SD of triplicate experiments was plotted ( n = 3), * p < 0.05.

Article Snippet: Ovarian carcinoma cell lines SKOV3 and A2780 were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Control, shRNA, Knockdown, Colony Assay, Flow Cytometry

Journal: Cancer Medicine

Article Title: BIRC5 facilitates cisplatin‐chemoresistance in a m 6 A ‐dependent manner in ovarian cancer

doi: 10.1002/cam4.6811

Figure Lengend Snippet: Upregulation of BIRC5 promotes cisplatin resistance of OC cells. (A) SKOV3 and A2780 cells transfected with overexpressed‐BIRC5 vector, and the effect were tested by RT‐qPCR and Western blotting. (B–D) CCK8 and colony formation assay revealed the cell viability of overexpression BIRC5 cells treated with 20 μM cisplatin, and the sensitivity to cisplatin of overexpressed BIRC5 cells measured by IC 50 . (E,F) Flow cytometry assays were performed to observe the change of percentage of apoptosis cells (early + late) after upregulating BIRC5, and the expression level of apoptosis biomarker PARP was detected by Western blotting. The mean ± SD of triplicate experiments was plotted ( n = 3), * p < 0.05.

Article Snippet: Ovarian carcinoma cell lines SKOV3 and A2780 were obtained from American Type Culture Collection (ATCC).

Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Colony Assay, Over Expression, Flow Cytometry, Expressing, Biomarker Discovery

Journal: Cancer Medicine

Article Title: BIRC5 facilitates cisplatin‐chemoresistance in a m 6 A ‐dependent manner in ovarian cancer

doi: 10.1002/cam4.6811

Figure Lengend Snippet: BIRC5 is regulated by METTL3‐mediated m 6 A modification. (A) RNA extracted in SKOV3, SKOV3/DDP, A2780 and A2780/DDP, and m6A status was examined by 450 nm absorbance according to m 6 A methylation assay kit manuscript (Abcam, USA). (B) Overexpression of predicted writers and measure the expression of BIRC5 in mRNA and protein level. (C) The mRNA and protein expression level of METTL3 in OC cells. (D–F) The effect of silenced METTL3, with the decreased BIRC5 expression level and m 6 A level. (G) MeRIP‐seq confirmed the m 6 A peak location. (H)Predicted binding motif of METTL3 on 3′UTR of BIRC5 mRNA from RMBase website. The mean ± SD of triplicate experiments was plotted ( n = 3), * p < 0.05.

Article Snippet: Ovarian carcinoma cell lines SKOV3 and A2780 were obtained from American Type Culture Collection (ATCC).

Techniques: Modification, Methylation, Over Expression, Expressing, Binding Assay

Journal: Cancer Medicine

Article Title: BIRC5 facilitates cisplatin‐chemoresistance in a m 6 A ‐dependent manner in ovarian cancer

doi: 10.1002/cam4.6811

Figure Lengend Snippet: METTL3/IGF2BP1/BIRC5 axis is essential for cisplatin resistance of OC. (A) The expression level of BIRC5 in silenced IGF2BP1 OC cells. (B–D) Cell viability detected by CCK8, the sensitivity of cisplatin detected by IC 50 , and the clonogenic ability in silenced IGF2BP1 OC cells (left panel), quantification of the colony formation assay results (right panel). (E) The mRNA expression of BIRC5 was measured by RT‐qPCR in SKOV3/DDP and A2780/DDP, who overexpressed or silenced IGF2BP1 upon actinomycin D (ActD) treatment for 8 h. (F–I) Nude mice bearing SKOV3/DDP cells treated with shBIRC5, or shBIRC5 combined with pcDNA‐IGF2BP1 plasmid (IGF2BP1) or corresponding negative control, and after 7 days the mice treated with cisplatin (10 mg/kg). Tumor volume were measured and record 4 days after injection. Tumor were removed after 28 days, and their weights had significantly differences. The protein expression of PARP was measured by Western blotting. The mean ± SD of triplicate experiments were plotted ( n = 3), * p < 0.05,** p < 0.01.

Article Snippet: Ovarian carcinoma cell lines SKOV3 and A2780 were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Colony Assay, Quantitative RT-PCR, Plasmid Preparation, Negative Control, Injection, Western Blot