cdk4 6 inhibitor ly2835219  (MedChemExpress)

Journal: Cancers

Article Title: Dual Targeting of CDK4/6 and cMET in Metastatic Uveal Melanoma

doi: 10.3390/cancers13051104

Figure Lengend Snippet: Abemaciclib induces G1 arrest and decreases cell growth in metastatic uveal melanoma cells. ( A ) UM001, UM002B, and UM004 cells were treated with DMSO or different concentrations of abemaciclib as indicated for 48 h. Cell lysates were probed with phospho-retinoblastoma (RB), total RB, cyclin A2, FOXM1, cyclin D1, CDK4, and β-actin antibodies. ( B ) Cells were treated with DMSO or 1 μM abemaciclib for 24 h. Cells were then fixed, permeabilized, and subjected to PI staining. Cell-cycle analysis was performed with FlowJo software. ** p < 0.01, based on the two sample t -test with unequal variance. ( C ) UM001, UM002B, and UM004 cells were treated with DMSO or different concentrations of abemaciclib as indicated. After seven days, cells were subjected to crystal violet staining. Representative microscopic images of the cells at ×200 magnification are shown. Scale bar, 100 μm. ( D ) UM001, UM002B, and UM004 cells were seeded in 96-well plates overnight and then treated with increasing doses of abemaciclib for 96 h. Cell viability was analyzed by the MTS assay. GI 50 values were calculated using Graph pad Prism. ( E ) UM001, UM002B, and UM004 cells were treated with DMSO or different concentrations of abemaciclib as indicated for 48 h. Cell lysates were probed with PARP and β-actin antibodies. Stau, UM004 cells were treated with 0.5 μM of staurosporine for 3 h as a positive control.

Article Snippet: A CDK4/6 inhibitor, Abemaciclib (LY2835219), and a cMET inhibitor, merestinib (LY2801653), were provided by Eli Lilly and Company, Indianapolis, IN, USA.

Techniques: Staining, Cell Cycle Assay, Software, MTS Assay, Positive Control

Journal: Cancers

Article Title: Dual Targeting of CDK4/6 and cMET in Metastatic Uveal Melanoma

doi: 10.3390/cancers13051104

Figure Lengend Snippet: The phosphorylation of RB in metastatic uveal melanoma and the effect of abemaciclib on inhibiting phosphorylated RB in metastatic uveal melanoma (UM) explants. ( A ) Representative images (magnification, ×400) of phospho-RB staining (red color arrow) illustrating the progressively increasing staining scores for metastatic samples of uveal melanoma patients. Staining positivity was scored as 0, negative; 1+, positive staining in <1% of tumor cells; 2+, in 1% to 10% of tumor cells; 3+, in >10% of tumor cells. ( B ) Fresh UM tumors obtained from surgery were cut into approximately 1 mm 3 pieces and plated on Vetspon absorbable hemostatic gelatin sponges for 48 h. The sponges were pre-soaked in medium containing DMSO as a vehicle control and 1 μM abemaciclib. Medium was replenished every 24 h. Surgical specimen and tumor pieces treated as above were formalin fixed and paraffin embedded. Tissue sections were stained with phospho-RB S807/811. Representative images (magnification, ×400) of phospho-RB staining (red color arrows) are shown.

Article Snippet: A CDK4/6 inhibitor, Abemaciclib (LY2835219), and a cMET inhibitor, merestinib (LY2801653), were provided by Eli Lilly and Company, Indianapolis, IN, USA.

Techniques: Phospho-proteomics, Staining, Control

Journal: Cancers

Article Title: Dual Targeting of CDK4/6 and cMET in Metastatic Uveal Melanoma

doi: 10.3390/cancers13051104

Figure Lengend Snippet: Hepatocyte growth factor (HGF) reduces the growth-inhibitory effect of abemaciclib in metastatic uveal melanoma cells. ( A ) Cells were exposed to various concentrations of abemaciclib as indicated with or without 10 ng/mL of HGF for 96 h and cell survival was assessed by the MTS assay. Data are presented as mean ± SD. ( B ) Cells were exposed to various concentrations of abemaciclib as indicated with or without 10 ng/mL of HGF for 48 h. Cells were lysed and phosphorylation of cMET, RB, AKT, ERK, and p70S6K was assessed by Western immunoblotting.

Article Snippet: A CDK4/6 inhibitor, Abemaciclib (LY2835219), and a cMET inhibitor, merestinib (LY2801653), were provided by Eli Lilly and Company, Indianapolis, IN, USA.

Techniques: MTS Assay, Phospho-proteomics, Western Blot

Journal: Cancers

Article Title: Dual Targeting of CDK4/6 and cMET in Metastatic Uveal Melanoma

doi: 10.3390/cancers13051104

Figure Lengend Snippet: Merestinib overcomes HGF-mediated resistance to abemaciclib in metastatic uveal melanoma cells. ( A ) UM002B and UM004 cells were treated with merestinib for 4 h, followed by 10 ng/mL of HGF stimulation for 15 min. Phosphorylation of cMET was evaluated by Western immunoblotting of cell lysates with phospho-cMET antibody. Actin was used as the loading control. ( B ) UM002B and UM004 cells were treated with DMSO or 1 μM abemaciclib, in combination with 10 ng/mL of HGF and/or 0.2 μM merestinib for 4 days. Cell viability was determined by the MTS assay. Data are presented as mean ± SD. ** p < 0.01, based on the two sample t -test with unequal variance. ( C ) UM002B and UM004 cells were treated with DMSO or 1 μM abemaciclib, in combination with 10 ng/mL of HGF and/or 0.2 μM of merestinib for seven days. Cells were stained and representative images are shown. ( D ) UM002B and UM004 cells were pretreated with DMSO, 1 μM of abemaciclib, or 0.2 μM of merestinib for 48 h. Cells were then stimulated with 10 ng/mL HGF for 1 h, as indicated. Activation of cMET, RB, AKT, ERK1/2, and p70S6K was analyzed by Western immunoblotting.

Article Snippet: A CDK4/6 inhibitor, Abemaciclib (LY2835219), and a cMET inhibitor, merestinib (LY2801653), were provided by Eli Lilly and Company, Indianapolis, IN, USA.

Techniques: Phospho-proteomics, Western Blot, Control, MTS Assay, Staining, Activation Assay

Journal: Cancers

Article Title: Dual Targeting of CDK4/6 and cMET in Metastatic Uveal Melanoma

doi: 10.3390/cancers13051104

Figure Lengend Snippet: HGF prevents abemaciclib induced-cell senescence and merestinib reverses HGF effects. ( A ) UM002B and UM004 cells were treated with DMSO or 1 μM of abemaciclib, in combination with 10 ng/mL of HGF and/or 0.2 μM of merestinib for six days. Cells were fixed and stained overnight for senescence-associated β-galactosidase activity. Positive cells were shown with arrows. Data are presented as mean ± SD. *, p < 0.05; **, p < 0.01, based on Tukey analysis. ( B ) UM002B and UM004 cells were treated with 1 μM of abemaciclib, in combination with 10 ng/mL of HGF and/or 0.2 μM of merestinib for 72 h. Activation of cMET, RB, FOXM1, AKT, ERK1/2, and p70S6K was analyzed by Western immunoblotting. ( C ) UM004 cells were transfected with control or FOXM1 targeting siRNA (#1 and #2) and cultured with 1 μM of abemaciclib in the presence or absence of 10 ng/mL of HGF for six days. Following six-day treatment, cells were fixed and stained overnight for senescence-associated b-galactosidase activity. Positive cells were shown with arrows. Knockdown of FOXM1 reversed the evasion of cellular senescence mediated by HGF. ( D ) UM004 cells were transfected with control or FOXM1 targeting siRNA (#1 and #2) in the presence of 1 μM of abemaciclib and 10 ng/mL of HGF for 72 h. FOXM1 and β-actin were analyzed by Western immunoblotting.

Article Snippet: A CDK4/6 inhibitor, Abemaciclib (LY2835219), and a cMET inhibitor, merestinib (LY2801653), were provided by Eli Lilly and Company, Indianapolis, IN, USA.

Techniques: Staining, Activity Assay, Activation Assay, Western Blot, Transfection, Control, Cell Culture, Knockdown

Journal: Cancers

Article Title: Dual Targeting of CDK4/6 and cMET in Metastatic Uveal Melanoma

doi: 10.3390/cancers13051104

Figure Lengend Snippet: Effects of the abemaciclib and merestinib combination on a metastatic uveal melanoma xenograft tumor model in human HGF knockin mice. ( A ) Mice bearing subcutaneous UM004 tumors were dosed with vehicle ( n = 5), 50 mg/kg abemaciclib ( n = 5), 12 mg/kg merestinib ( n = 5), or abemaciclib plus merestinib combination ( n = 5) once daily for 28 days. Tumor size was measured by caliper twice weekly. ( B ) Tumor weight. Data represent mean volume ± SD. * p < 0.05; ** p < 0.01, based on Dunnett’s test. ( C ) Average percentage change in body weight after treatments. ( D ) UM004 tumors were excised from mice following seven-day treatment. Tumor lysates were prepared and analyzed by Western immunoblotting using phospho-cMET, cMET, phospho-RB, total RB, FOXM1, and β-actin. ( E ) Representative images of immunohistochemical staining for hematoxylin and eosin (H & E), IgG isotype control, phospho-cMET, phospho-RB, FOXM1, and Ki67 in UM004 tumor xenografts. Original magnification, ×400. Scale bar, 50 μm.

Article Snippet: A CDK4/6 inhibitor, Abemaciclib (LY2835219), and a cMET inhibitor, merestinib (LY2801653), were provided by Eli Lilly and Company, Indianapolis, IN, USA.

Techniques: Knock-In, Western Blot, Immunohistochemical staining, Staining, Control

Journal: The American Journal of Pathology

Article Title: Cyclin-Dependent Kinase Inhibitors and the Treatment of Gastrointestinal Cancers

doi: 10.1016/j.ajpath.2015.01.008

Figure Lengend Snippet: Key regulators of the mammalian cell cycle. The green plus signs represent positive regulators of cell cycle progression, whereas the red minus sign are cell cycle inhibitory proteins. The yellow P represent phosphorylation events on the Rb. Also shown are three CDK4/6 inhibitors that are currently in various stages of clinical development: PD-033299 (Pfizer, New York, NY), LY2835219 (Eli Lilly, Indianapolis, IN), and LEE011 (Novartis, Basel, Switzerland). CDK, cyclin-dependent kinase; Rb, retinoblastoma protein.

Article Snippet: 73 Of note, other CDK4/6 inhibitors, LY2835219 (Eli Lilly, Indianapolis, IN) and LEE011 (Novartis, Basel, Switzerland), 74 , 75 , 76 have also entered clinical development and have shown promising activity in breast cancer, lung cancer, and other solid tumors.

Techniques: Phospho-proteomics

Journal: The American Journal of Pathology

Article Title: Cyclin-Dependent Kinase Inhibitors and the Treatment of Gastrointestinal Cancers

doi: 10.1016/j.ajpath.2015.01.008

Figure Lengend Snippet: Cyclins, CDKs, and Their Inhibitors

Article Snippet: 73 Of note, other CDK4/6 inhibitors, LY2835219 (Eli Lilly, Indianapolis, IN) and LEE011 (Novartis, Basel, Switzerland), 74 , 75 , 76 have also entered clinical development and have shown promising activity in breast cancer, lung cancer, and other solid tumors.

Techniques: