egfr/her2 inhibitor lapatinib  (LC Laboratories)

Journal: Advanced Science

Article Title: Osteoblast‐Derived ECM1 Promotes Anti‐Androgen Resistance in Bone Metastatic Prostate Cancer

doi: 10.1002/advs.202407662

Figure Lengend Snippet: ECM1 activates the MAPK signaling pathway in PCa cells. A) Waterfall plot showing differentially expressed genes (p < 0.05, log2 fold change>1.5) in C4‐2B cells treated with ENZ (10 µM, 48 h) combined with ECM1 protein (200 ng mL −1 , 48 h) or ENZ (10 µM, 48 h) alone. The highlighted genes were related to proliferation and apoptosis. B) KEGG analysis of pathways enriched in ENZ combined with ECM1 protein treatment group compared to the ENZ treatment group. C) WB analysis of MEK, p‐MEK, ERK1/2, and p‐ERK1/2 expression in the indicated groups of C4‐2B cells. D) IHC staining and quantification of p‐ERK1/2 expression in mice intratibial and subcutaneous tumors grouped as indicated (Scale bars, 500 µm and 100 µm, n = 6 per group). E) WB analysis and quantification of MEK, p‐MEK, ERK1/2, and p‐ERK1/2 expression in C4‐2B cells stimulated with ENZ (10 µM) combined with ECM1 (200 ng mL −1 ) in the presence of inhibitors for either RAS (MCP110, 10 µM), MEK (U0126, 10 µM), ERK1/2 (Ulixertinib, 10 µM), EGFR (Lapatinib, 10 µM), FGFR1 (Fexagratinib, 10 µM), IGF1R (Linsitinib, 10 µM) or Veh (DMSO), compared to ENZ‐treated alone or untreated C4‐2B cells. F) C4‐2B cell proliferation on day 7 of groups as shown in E. G) WB analysis and quantification of EGFR, p‐EGFR, FGFR1, p‐FGFR1, IGF1R and p‐IGF1R expression in groups as indicated. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: The Ras inhibitor MCP110 (MCE; HY‐123673), MEK inhibitor U0126 (MCE; HY‐12031A), ERK inhibitor ulixertinib (MCE; HY‐15816), EGFR inhibitor lapatinib (MCE; HY‐50898), FGFR1 inhibitor fexagratinib (MCE; HY‐13330), and IGF1R inhibitor linsitinib (MCE; HY‐10191) required dissolution in DMSO before use.

Techniques: Expressing, Immunohistochemistry

Journal: Cancer research

Article Title: Dual Inhibition of SYK and EGFR Overcomes Chemoresistance by Inhibiting CDC6 and Blocking DNA Replication.

doi: 10.1158/0008-5472.CAN-24-0769

Figure Lengend Snippet: Figure 1. Mechanism and effect of dual TKI treatment in ovarian cancer cell lines. A, Western blotting of the indicated molecules in two cell lines, KOC7C and SKOV3, after treatment with SYK inhibitor (R406) and EGFR inhibitor (lapatinib), either alone or in combination. Combo+ (left, fifth lane) indicates a higher dose of R406 (8 μmol/L) and lapatinib (10 μmol/L). β-Actin was used as a loading control. B and C, Brightfield images of the indicated cell lines 48 hours after treatment at the concentrations shown in C. D and E, Colony formation assay after knocking down SYK and/or EGFR. The knockdown efficiency is presented in Supplementary Fig. S1A. F, Western blot analysis of EGFR and SYK in the indicated ovarian cancer cell lines for cell model selection. β-Actin was used as a loading control. *, cultures from xenografts. G–I, Colony formation assay in KOC7C and OVCA429 cells using the combination of increasing doses of the drugs (R, R406; L, lapatinib). The heatmap shows the relative number of colonies. Cell colonies are presented in Supplementary Fig. S1D and S1E. The drug interactions for these combinations are plotted as CDI values (I). J–M, Fa–CI plot of the indicated cell lines after treatment with R406 (0–5 μmol/L) or lapatinib (0–5 μmol/L). The CDI plot for this assay is shown in Supplementary Fig. S1F. Both drugs were serially diluted from 0 to 5 μmol/L (Supplementary Fig. S1G and S1H). ∗, P < 0.05; ∗∗, P < 0.01.

Article Snippet: SYK inhibitors (R406, #S2194; entospletinib, #GS-9973) and EGFR inhibitors (lapatinib, #S1028; erlotinib, #S1023), along with ERK1/ 2 inhibitors (ulixertinib, #S7854; temuterkib, #S8534), were purchased from Selleckchem.

Techniques: Western Blot, Control, Colony Assay, Knockdown, Selection

Journal: Cancer research

Article Title: Dual Inhibition of SYK and EGFR Overcomes Chemoresistance by Inhibiting CDC6 and Blocking DNA Replication.

doi: 10.1158/0008-5472.CAN-24-0769

Figure Lengend Snippet: Figure 4. Elucidation of CDC6 regulatory mechanisms after combined TK inhibition. A, qPCR was used to evaluate the mRNA expression of CDC6 after drug treatment (DMSO, R406, lapatinib, or a combination of both) in KOC7C and SKOV3 cells. *, P < 0.05; **, P < 0.001; ***, P < 0.0001 (Student t test). B, IPA-derived “upstream regulator” analysis depicting the regulatory mechanisms impacting CDC6 expression. Proteins that had a z score ± 2 and P < 0.05 were considered significant. C, Western blot validation of downregulated E2F1 after drug treatment (DMSO, R406, lapatinib, or a combination of both). D, Analysis of the effects of erlotinib (an EGFR inhibitor) and entospletinib (an SYK inhibitor) on E2F1 and CDC6 expression by Western blotting. E and F, Fa–CI plot of the indicated cell lines after treatment with entospletinib (0–5 μmol/L) or erlotinib (0–5 μmol/L). G, Western blot analysis showing that ulixertinib and temuterkib, ERK1/2–MAPK inhibitors, affect the protein expression of pERK1/2, ERK1/2, pCDC6 (Ser54), CDC6, and E2F1 in KOC7C, SKOV3, and OVCAR3 cell lines. H, Representative brightfield images of PDOs in different treatment groups. I and J, Fa–CI plot of the indicated PDOs after treatment with entospletinib (0–5 μmol/L) or erlotinib (0– 5 μmol/L). K, Schematic representation showing the regulatory dynamics wherein the EGFR–ERK pathway modulates E2F1, leading to the transcriptional control of various genes, including CDC6 and MCM.

Article Snippet: SYK inhibitors (R406, #S2194; entospletinib, #GS-9973) and EGFR inhibitors (lapatinib, #S1028; erlotinib, #S1023), along with ERK1/ 2 inhibitors (ulixertinib, #S7854; temuterkib, #S8534), were purchased from Selleckchem.

Techniques: Inhibition, Expressing, Derivative Assay, Western Blot, Biomarker Discovery, Control