Journal: The EMBO Journal

Article Title: Cyclophilin D plays a critical role in the survival of senescent cells

doi: 10.1038/s44318-024-00259-2

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Abemaciclib , MedChem Express , HY-16297.

Techniques: Recombinant, Sequencing, Diagnostic Assay, Staining, Cell Culture, Membrane, Western Blot, Transfection, DC Protein Assay, Software, Cell Viability Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting PEG10 as a novel therapeutic approach to overcome CDK4/6 inhibitor resistance in breast cancer

doi: 10.1186/s13046-023-02903-x

Figure Lengend Snippet: Ectopic overexpression of PEG10 augments EMT and leads to palbociclib resistance. A Immunoblot demonstrates ectopic overexpression of different PEG10 protein isoforms in MCF7 and T47D cells. Ectopic overexpression of PEG10 elevated the ZEB1 and suppressed the E-cadherin expression. B , C Representative images from a migration and invasion assay after ectopic overexpression of different PEG10 protein isoforms in MCF7 and T47D cells. The area of migratory and invading cells from three different non-overlapping 100 × microscopic fields is expressed as mean ± SD in the right panel. Independent sample t-test: * p < 0.05, ** p < 0.01, *** p < 0.001. D - I Cell viability (MTT) assay of MCF7 and T47D cells before and after ectopic overexpression of PEG10-RF1 and subsequent treatment with the indicated dose of palbociclib, abemaciclib, and ribociclib for 72 h. Three independently repeated experiments were performed with similar results. Independent sample t-test: * p < 0.05, ** p < 0.01, *** p < 0.001, Abbreviation: ns, not significant. J , K Cell cycle distribution of MCF7 and T47D cell line before and after the ectopic overexpression of different PEG10 protein isoforms and subsequent treatment with the IC 50 concentration of palbociclib for 48 h. Three independently repeated experiments were performed with similar results. Independent sample t-test: ** p < 0.01, Abbreviation: ns, not significant

Article Snippet: Palbociclib (cat# CS-0019), abemaciclib (cat# CS-1230) and ribociclib (cat# CS-1750) were purchased from ChemScene (Monmouth Junction, NJ, USA).

Techniques: Over Expression, Western Blot, Expressing, Migration, Invasion Assay, MTT Assay, Concentration Assay

Journal: Cancer discovery

Article Title: The genomic landscape of intrinsic and acquired resistance to cyclin-dependent kinase 4/6 inhibitors in patients with hormone receptor positive metastatic breast cancer

doi: 10.1158/2159-8290.CD-19-1390

Figure Lengend Snippet: (a) T47D cells were modified via CRISPR-mediated downregulation (RB1) or lentiviral overexpression (AKT1, KRAS G12D, AURKA, CCNE2) to interrogate potential resistance mediators identified in patient biopsy samples. Western blotting with the indicated antibodies is included. (b-f) Modified T47D cells were exposed to escalating doses of CDK4/6i (palbociclib – left, abemaciclib – right) and viability was estimated via cell-titer-glo (CTG) assay. Control (CRISPR non-targeting guide or GFP) cells are plotted along with the resistance driver of interest (RB1 – b, AKT1 – c, KRAS G12D – d, AURKA – e, CCNE2 – f). Parental and variant cell lines are normalized to vehicle control and viability is plotted as a function of increasing CDK4/6i (graphed as triplicate average +/− standard deviation). All variants provoke CDK4/6i resistance (to both palbociclib and abemaciclib) in vitro in T47D cells. Corresponding IC50 values are included in Supplemental Table 7.

Article Snippet: Chemicals utilized included palbociclib (Selleck Chemicals, S1116), abemaciclib (ApexBio, A1794), and fulvestrant (Sigma-Aldrich, I4409).

Techniques: Modification, CRISPR, Over Expression, Western Blot, CTG Assay, Variant Assay, Standard Deviation, In Vitro

Journal: Cancer discovery

Article Title: The genomic landscape of intrinsic and acquired resistance to cyclin-dependent kinase 4/6 inhibitors in patients with hormone receptor positive metastatic breast cancer

doi: 10.1158/2159-8290.CD-19-1390

Figure Lengend Snippet: (a) Breast cancer cell lines (T47D, MCF7, MDA-MB-361) were cultured long-term to resistance in the presence of CDK4/6i (palbociclib, abemaciclib). The resulting cell lines which emerged were subjected to western blotting for putative mediators of drug resistance (RB1, AKT1, KRAS/ERK, AURKA, and CCNE2). (b-c) T47D cells cultured to resistance in the presence of abemaciclib demonstrated low levels of RB1 expression (T47D-AR1) and increased sensitivity to the AURKA inhibitor LY3295668. MDA-MB-361 cells cultured to resistance in the presence of abemaciclib demonstrated high levels of ERK activation (361-AR1) and increased sensitivity to the ERK inhibitor LY3214996. MDA-MB-361 cells cultured to resistance in the presence of palbociclib demonstrated high levels of AURKA (361-PR1) and increased sensitivity to the AURKA inhibitor LY3295668. MCF7 cells cultures to resistance in the presence of palbociclib demonstrated increased levels of CCNE2 (MCF7-PR1) and increased sensitivity to the CHEK1 inhibitor prexasertib.

Article Snippet: Chemicals utilized included palbociclib (Selleck Chemicals, S1116), abemaciclib (ApexBio, A1794), and fulvestrant (Sigma-Aldrich, I4409).

Techniques: Cell Culture, Western Blot, Expressing, Activation Assay

Journal: eLife

Article Title: Proliferative exhausted CD8 + T cells exacerbate long-lasting anti-tumor effects in human papillomavirus-positive head and neck squamous cell carcinoma

doi: 10.7554/eLife.82705

Figure Lengend Snippet: ( a ) Single-cell transcriptomic profiling of HNSCC tumor microenvironment (TME). Twenty cell clusters are identified, colored by cell types. ( b ) The proliferation status of P-Tex and epithelial cells in violin plot. ( c ) The kernel density estimate distribution of proliferation markers (CDK4 and MKI67) and epithelial cancer cell markers (KRT15 and CD24) in uniform manifold approximation and projection (UMAP) plots. ( d ) The overall survival rate of HPV + /HPV - HNSCC patients in The Cancer Genome Atlas (TCGA) cohort related to the expression levels of CDK4 gene, adjusted for age and gender. ( e ) The proportion of P-Texs, T-Tex, and TEFF clusters in HPV + and HPV - samples in TCGA cohort by using the deconvolution algorithm; statistics were assessed by Chi-square tests. Marker genes that were used to define cell clusters in ( a ) are deconvolved into the TCGA data to obtain the proportion of P-Texs, T-Tex, and Teff clusters in the TCGA cohort. ( f ) The cell viability of P-Tex and cancer epithelial cells assessed by CCK8 experiment after Abemaciclib treated in vitro. ***: p<0.001, **: p<0.01, *: p<0.05.

Article Snippet: Chemical compound, drug , Abemaciclib (LY2835219) , Topscience , Cat#T2381 , .

Techniques: Expressing, Marker, In Vitro

Journal: eLife

Article Title: Proliferative exhausted CD8 + T cells exacerbate long-lasting anti-tumor effects in human papillomavirus-positive head and neck squamous cell carcinoma

doi: 10.7554/eLife.82705

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , Abemaciclib (LY2835219) , Topscience , Cat#T2381 , .

Techniques: Sequencing, CCK-8 Assay, Software

Journal: Oncogene

Article Title: Lysosomal trapping of palbociclib and its functional implications

doi: 10.1038/s41388-019-0695-8

Figure Lengend Snippet: Lysosomal trapping is a feature common to all CDK4 inhibitors. a Donor SK-Mel-103 cells were treated with 1 μM palbociclib, 1 μM abemaciclib or 1 μM ribociclib for 7 days. The drugs were added only once and the culture media were not changed for the length of the treatment. Following treatments, cells were tripsinized and identical numbers were seeded in new dishes in the presence of the drugs. 24 h later the dishes were washed with PBS and incubated with fresh media for 3 additional days. The media were then collected, filtrated with 0.45 μM filters, diluted 1:2 with fresh media, and added to the recipient cells. Recipient SK-Mel-103 were incubated with the conditioned media for 7 days, with no media changes, and then stained for SA-βgal activity. b The reversible storage of palbociclib into lysosomes (left) has three main implications: (top right) it prolongs the availability of the drug (long-term activity) and induces paracrine effects (paracrine activity); (bottom right) the presence of another lysosomotropic drug, reduces the accumulation of palbociclib into lysosomes and this increases its cytoplasmic concentration thereby potentiating its intracellular activity (enhanced activity) as well as its extracellular release (paracrine activity)

Article Snippet: For senescence induction SK-Mel-103 cells were treated with 1–4 μM palbociclib (Selleckchem), 1 μM ribociclib (Selleckchem), 1 μM abemaciclib (Absource Diagnostics GmbH), 10 nM doxorubicin (Sigma), bleomycin 12 mUnits/ml (Selleckchem), 10 μM nutlin-3 (Sigma), or γ-irradiated (10 Gy).

Techniques: Incubation, Staining, Activity Assay, Concentration Assay

Journal: Frontiers in Oncology

Article Title: CDK4/6 inhibitors versus PI3K/AKT/mTOR inhibitors in women with hormone receptor-positive, HER2-negative metastatic breast cancer: An updated systematic review and network meta-analysis of 28 randomized controlled trials

doi: 10.3389/fonc.2022.956464

Figure Lengend Snippet: Baseline characteristics of 28 RCTs.

Article Snippet: next MONARC(b) ( ) , Erika Hamilton et al. 2021.06 , II , Abemaciclib 150 mg (79) , Abemaciclib200 mg + loperamide (77) , NA , NA , NA , NA , 62 , 62.3.

Techniques: