786-o parental  (Broad Institute Inc)

Journal: Cancer Genomics & Proteomics

Article Title: Timescale-dependent Phosphoproteomic Remodeling and Motility-associated Adaptation under Chronic Cabozantinib Exposure in Renal Cell Carcinoma

doi: 10.21873/cgp.20576

Figure Lengend Snippet: Growth‑inhibitory effect of cabozantinib. (A) Dose-response cell viability after 48 h cabozantinib treatment in 786-O and Caki-1 cells. Values represent mean ± standard deviation (SD) (n=4); *p<0.01 vs. vehicle. (B) Immunoblot analysis of total MET and phospho-MET (Y1234/1235) in parental (Par) and chronically treated (Chr) cells, with or without cabozantinib treatment. Cabozantinib robustly suppressed phospho-MET in both Par and Chr states of both cell lines. β-actin serves as a loading control. (C) Time-course viability (24-72 h) of Par and Chr 786-O cells. (D) Time-course viability (24-72 h) of Par and Chr Caki-1 cells. Cab: acute cabozantinib-treated; Par: parental; Chr: chronically cabozantinib-treated cells (>4 months). *p<0.05; **p<0.01. Statistical significance was determined using an unpaired two-tailed Student’s t-test.

Article Snippet: Parental 786-O and Caki-1 renal cell carcinoma cell lines were obtained from ATCC (Manassas, VA, USA) and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Standard Deviation, Western Blot, Control, Two Tailed Test

Journal: Cancer Genomics & Proteomics

Article Title: Timescale-dependent Phosphoproteomic Remodeling and Motility-associated Adaptation under Chronic Cabozantinib Exposure in Renal Cell Carcinoma

doi: 10.21873/cgp.20576

Figure Lengend Snippet: Quantitative phosphoproteomic profiling of acute and chronic cabozantinib exposure in 786-O cells. (A) Volcano plot showing significantly regulated phosphosites in the Cab/Par comparison after 48 h acute treatment (FDR <0.01, s0=0.1). Red and green dots indicate significantly upregulated and downregulated phosphosites, respectively; grey dots indicate non-significantly regulated phosphosites. (B) Volcano plot showing significantly regulated phosphosites in the Chr/Par comparison after >4 months of chronic exposure (FDR <0.01, s0=0.1). Color coding is as in (A). (C) Volcano plot directly comparing the two drug-treated states (Chr vs. Cab), highlighting differentially regulated phosphosites using the same color scheme (D) KEGG pathway enrichment for phosphoproteins significantly regulated in the Cab/Par condition. (E) Reactome pathway enrichment for phosphoproteins significantly regulated in the Cab/Par condition. (F) KEGG pathway enrichment for phosphoproteins significantly regulated in the Chr/Par condition. (G) Reactome pathway enrichment for phosphoproteins significantly regulated in the Chr/Par condition. Phosphosite regulation was determined from log2-transformed ratio using permutation-based statistics. Enrichment analyses were performed separately for Cab/Par and Chr/Par contrasts. Pathway enrichment analyses were performed using significantly regulated phosphosites as the input list, with all quantified phosphoproteins used as the background set.

Article Snippet: Parental 786-O and Caki-1 renal cell carcinoma cell lines were obtained from ATCC (Manassas, VA, USA) and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Comparison, Phospho-proteomics, Transformation Assay

Journal: Cancer Genomics & Proteomics

Article Title: Timescale-dependent Phosphoproteomic Remodeling and Motility-associated Adaptation under Chronic Cabozantinib Exposure in Renal Cell Carcinoma

doi: 10.21873/cgp.20576

Figure Lengend Snippet: Migration, invasion, and signaling adaptations in 786-O cells under chronic cabozantinib exposure. (A) Transwell migration assay of Par and Chr 786-O cells. Migrated cells were fixed, crystal-violet stained, and quantified using ImageJ. (B) Matrigel invasion assay performed using the same cell model and quantification workflow as in (A). Invasion was assessed following 6 h incubation through Matrigel-coated inserts. (C) Immunoblot analysis of selected signaling nodes in Par and Chr 786-O cells, including MET, phospho-MET (Y1234/1235), ERK1/2, phospho-ERK, c-Jun, phospho-c-Jun (S63), HSPB1, and phospho-HSPB1 (S82). β-actin served as loading control. (D) Network representation of Chr-associated phosphosites mapped to adhesion- and stress-associated signaling modules based on the annotation-enrichment analysis. Data are shown as mean ± standard deviation (SD) from three independent experiments. Statistical significance was determined using an unpaired two-tailed Student’s t-test; #p<0.05; *p<0.01.

Article Snippet: Parental 786-O and Caki-1 renal cell carcinoma cell lines were obtained from ATCC (Manassas, VA, USA) and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Migration, Transwell Migration Assay, Staining, Invasion Assay, Incubation, Western Blot, Control, Standard Deviation, Two Tailed Test