Journal: Cancers

Article Title: Dynamic Phosphoproteomic Profiling Identifies Casein Kinase 2 as a Critical Survival Kinase in Quiescent Breast Cancer Cells and a Potential Therapeutic Target for Minimal Residual Disease

doi: 10.3390/cancers18091449

Figure Lengend Snippet: Validation of protein and phosphorylation changes in TNBC cell lines and patient data. ( A ) Expression and site-specific phosphorylation of indicated proteins was analyzed in MDA-MB-231, Hs578T and BT-549 in various timepoints corresponding to the transition between quiescence and proliferation. Timepoints were as follows: A —continuously proliferating cells; B —serum-starved cells (48 h); C —serum-starved cells (96 h); D —serum-reactivated cells (+20 min); E —serum-reactivated cells (+120 min). α-tubulin was used as loading control. ( B , C ) Analysis of site-specific phosphorylation and protein expression of indicated proteins in CPTAC patient samples dataset. ( D ) Comparison table of top quiescence-downregulated proteins in vitro (screening conditions) and in CPTAC patients’ samples dataset. FC—fold change, ns—not significant, *, **, ***, and **** represent p -values < 0.05, <0.01, <0.001, and <0.0001. ( E ) Box plots showing relative protein expression levels derived from the CPTAC dataset for proteins significantly altered between MYC-amplified and MYC non-amplified breast tumors. ( F ) Comparison table of top quiescence-downregulated site-specific phosphorylations in vitro (screening conditions) and in CPTAC patients’ samples dataset. FC—fold change, ns—not significant, *, **, ***, and **** represent p -values < 0.05, <0.01, <0.001, and <0.0001. ( G ) Box plots showing relative site-specific phosphorylation levels derived from the CPTAC dataset for phosphosites significantly altered between MYC-amplified and MYC non-amplified breast tumors.

Article Snippet: MDA-MB-231, Hs578T and BT-549 were obtained from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Biomarker Discovery, Phospho-proteomics, Expressing, Control, Comparison, In Vitro, Derivative Assay, Amplification

Journal: Cancers

Article Title: Dynamic Phosphoproteomic Profiling Identifies Casein Kinase 2 as a Critical Survival Kinase in Quiescent Breast Cancer Cells and a Potential Therapeutic Target for Minimal Residual Disease

doi: 10.3390/cancers18091449

Figure Lengend Snippet: CK2 regulates DAPK3 to maintain survival under stress conditions. ( A ) Expression and cleavage of PARP1 was analyzed in MDA-MB-231, Hs578T and BT-549 under indicated conditions (+/− FBS, +/− CK2 inhibition) by Western blot. α-Tubulin was used as loading control. ( B ) TNBC cell lines (MDA-MB-231, Hs578T, BT-549) were treated with indicated concentrations of CX-4945 in serum containing as well as serum-free conditions and analyzed for cell viability using the MTT assay. Relative viability for each cell line relative to DMSO-treated cells at day 1 at respective condition is shown. ( C ) TNBC cell lines (MDA-MB-231, Hs578T, BT-549) were treated with indicated concentrations of CX-4945 and Doxorubicin and analyzed for cell viability using the MTT assay. Relative viability for each cell line relative to DMSO-treated cells at day 1 at respective condition is shown. ( D ) Volcano plot representing upregulated and downregulated proteins identified through phospho-CK2 substrate antibody pull-down after treatment of MDA-MB-231 with CX-4945. ( E ) Expression and cleavage of PARP1 was analyzed in MDA-MB-231 under indicated conditions (+/− FBS, +/− CK2 inhibition, +/− DAPK3 inhibition) by Western blot. α-Tubulin was used as loading control. ( F ) Bar chart representing quantification of PARP1 cleavage shown as ratio of full-length (FL) PARP1 to cleaved PARP1 from three independent experiments. ns—not significant. ( G ) Proposed model representing interaction of CK2 and DAPK3 in TNBC cells.

Article Snippet: MDA-MB-231, Hs578T and BT-549 were obtained from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Inhibition, Western Blot, Control, MTT Assay

Journal: medRxiv

Article Title: The miR-362-3p/ BCLAF1 axis regulates cisplatin sensitivity and metastatic progression in triple-negative breast cancer

doi: 10.64898/2026.03.09.26347941

Figure Lengend Snippet: A-C. Kaplan-Meier survival analysis of mice bearing MDA-MB-231 ( A ), CAL-51 ( B ), or MDA-MB-436 ( C ) tumors treated with cisplatin. Median survival was extended by miR-362-3p overexpression (OE) and shortened by miR-362-3p-inhibition compared to respective controls. D. Representative gross morphology of livers at necropsy; white arrows indicate macroscopic metastatic nodules in non-targeting controls (NTC) vs. miR-362-3p-OE. E. Quantitative of metastatic burden from H&E stained liver sections in MDA-MB-231 models. F. qRT-PCR validation of miR-362-3p levels in endpoint xenograft tumors, confirming sustained OE or inhibition throughout the study. Data represent mean±SD. *, ** and *** represent p <0.05, p <0.01, and p <0.001, respectively. Empty vector, EV.

Article Snippet: MDA-MB-231, MDA-MB-436, MDA-MB-468, and HCC1937 were acquired from the American Type Culture Collection (ATCC; Manassas, VA), CA51 (CVCL_1110) was obtained from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH (Braunschweig, Germany) and SUM149 which was a gift from Dr. Christina Gewinner, Division of Signal Transduction, BIDMC.

Techniques: Over Expression, Inhibition, Staining, Quantitative RT-PCR, Biomarker Discovery, Plasmid Preparation

Journal: medRxiv

Article Title: The miR-362-3p/ BCLAF1 axis regulates cisplatin sensitivity and metastatic progression in triple-negative breast cancer

doi: 10.64898/2026.03.09.26347941

Figure Lengend Snippet: A. Western blotting demonstrating that miR-362-3p overexpression (OE) suppresses BCLAF1 and further potentiates cisplatin-induced γ-H2AX accumulation in vitro . B. Western blot showing that miR-362-3p inhibition in MDA-MB-436 cells attenuates γ-H2AX expression following cisplatin treatment. C. Functional rescue of cisplatin sensitivity in miR-362-3p-inhibited cells following BCLAF1 knockdown using two independent shRNAs. Data represent mean±SD. # , ## and ### represent adj p <0.05, adj p <0.01, and adj p <0.001, respectively. Non-targeting controls, NTC; empty vector, EV.

Article Snippet: MDA-MB-231, MDA-MB-436, MDA-MB-468, and HCC1937 were acquired from the American Type Culture Collection (ATCC; Manassas, VA), CA51 (CVCL_1110) was obtained from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH (Braunschweig, Germany) and SUM149 which was a gift from Dr. Christina Gewinner, Division of Signal Transduction, BIDMC.

Techniques: Western Blot, Over Expression, In Vitro, Inhibition, Expressing, Functional Assay, Knockdown, Plasmid Preparation