mouse anti kras  (Santa Cruz Biotechnology)

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: H2122AR14 and H2122AR30 cells are resistant to sotorasib in vitro and show EGFR activation. A, Cell viability assay for KRAS G12C -mutant parental H2122 cells and H2122AR14 and H2122AR30 sotorasib-resistant clones exposed to the indicated concentrations of sotorasib for 72 hours. Data are means ± SD ( n = 3 independent experiments). B, Immunoblot analysis of KRAS, phosphorylated (p) and total (t) forms of AKT and ERK, and actin (loading control) in H2122, H2122AR14, and H2122AR30 cells treated with sotorasib (1 μmol/L) for 0 or 24 hours. C, Human phospho-RTK assay for H2122, H2122AR14, and H2122AR30 cells treated with 1 μmol/L sotorasib for 72 hours. D, Immunoblot analysis of Y845-, Y1068-, or Y1173-phosphorylated and total forms of EGFR in H2122 and H2122AR30 cells. Data in B–D are representative of at least three independent experiments.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: In Vitro, Activation Assay, Viability Assay, Mutagenesis, Clone Assay, Western Blot, Control

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: Combination treatment with EGFR and KRAS G12C inhibitors overcomes sotorasib resistance due to PTPRR downregulation. A, Cell viability assay for H2122 cells transfected with nonspecific control (siSCR) or PTPRR-specific (siPTPRR) siRNAs and then exposed to the indicated concentrations of sotorasib for 72 hours. B, Immunoblot analysis of PTPRR as well as total and phosphorylated forms of AKT and ERK in H2122 cells transfected with siRNAs as in ( A ) and then incubated with or without sotorasib (1 μmol/L) for 4 hours. C, Cell viability assay for H2122AR30 cells stably transfected with control (GFP) or PTPRR expression plasmids and treated with the indicated concentrations of sotorasib for 72 hours. D, Immunoblot analysis of total or phosphorylated forms of EGFR, AKT, and ERK in H2122AR30 cells treated with sotorasib (1 μmol/L), cetuximab (10 μg/mL), or the combination of both drugs for 4 hours. E, Colony formation assay for H2122 and H2122AR30 cells incubated with or without sotorasib (1 μmol/L), cetuximab (10 μg/mL), or both drugs for 4 weeks. Surviving cells were stained with crystal violet. Data in A and C are means ± SD ( n = 3 independent experiments), and those in B , D , and E are representative of at least three independent experiments.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: Viability Assay, Transfection, Control, Western Blot, Incubation, Stable Transfection, Expressing, Colony Assay, Staining

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: Expression of PTPRR is regulated by promoter DNA methylation in KRAS G12C -mutant NSCLC cell lines. A, The human PTPRR gene including the promoter regions for the long and short isoforms of the encoded protein is shown. The positions of CpG islands as well as the histone marks H3K4me1, H3K4me3, and H3K27ac are indicated. B, Abundance of transcripts for the long and short isoforms of PTPRR in tumor specimens from patients with NSCLC in the dbGaP database ( n = 73). Data are presented as dot plots with means ± SD indicated by the dashed and solid lines for the short isoform, and they were analyzed with the Wilcoxon ranked-sum test. C, RT-qPCR analysis of mRNA abundance for the short form of PTPRR in the indicated KRAS G12C -mutant cell lines. Data are presented as dot plots, with means + SD being indicated by the dashed and solid lines ( n = 3 independent experiments) and analyzed by one-way ANOVA and Dunnett’s test. D, Bisulfite genomic sequencing of the promoter region for the short variant of PTPRR in the indicated KRAS G12C -mutant cell lines. The percentage of methylated CpG sites among total CpG sites is shown. Data are presented as violin plots, with the median and upper quartile values indicated by the dashed and dotted lines and analyzed by one-way ANOVA and Dunnett’s test. Data are representative of three independent experiments.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: Expressing, DNA Methylation Assay, Mutagenesis, Quantitative RT-PCR, Genomic Sequencing, Variant Assay, Methylation

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: Clinical relevance of PTPRR expression to sotorasib treatment in individuals with NSCLC. A, RT-qPCR analysis of PTPRR mRNA (short form) abundance in normal lung tissue ( n = 9) and NSCLC tumor tissue ( n = 10). Data are presented as dot plots with means ± SD indicated by the dashed and solid lines, and they were analyzed with the Wilcoxon ranked-sum test. B, Representative IHC images for specimens of normal lung tissue (negative) and NSCLC tumor tissue (low and high) stained with antibodies to PTPRR. Scale bars, 100 μm. C and D, Kaplan–Meier analysis of PFS ( C ) and OS ( D ) according to low ( n = 5) or high ( n = 8) status for cytoplasmic PTPRR staining in tumor cells for patients with KRAS G12C -mutant NSCLC treated with sotorasib.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: Expressing, Quantitative RT-PCR, Staining, Mutagenesis

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: H2122AR14 and H2122AR30 cells are resistant to sotorasib in vitro and show EGFR activation. A, Cell viability assay for KRAS G12C -mutant parental H2122 cells and H2122AR14 and H2122AR30 sotorasib-resistant clones exposed to the indicated concentrations of sotorasib for 72 hours. Data are means ± SD ( n = 3 independent experiments). B, Immunoblot analysis of KRAS, phosphorylated (p) and total (t) forms of AKT and ERK, and actin (loading control) in H2122, H2122AR14, and H2122AR30 cells treated with sotorasib (1 μmol/L) for 0 or 24 hours. C, Human phospho-RTK assay for H2122, H2122AR14, and H2122AR30 cells treated with 1 μmol/L sotorasib for 72 hours. D, Immunoblot analysis of Y845-, Y1068-, or Y1173-phosphorylated and total forms of EGFR in H2122 and H2122AR30 cells. Data in B–D are representative of at least three independent experiments.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: In Vitro, Activation Assay, Viability Assay, Mutagenesis, Clone Assay, Western Blot, Control

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: Combination treatment with EGFR and KRAS G12C inhibitors overcomes sotorasib resistance due to PTPRR downregulation. A, Cell viability assay for H2122 cells transfected with nonspecific control (siSCR) or PTPRR-specific (siPTPRR) siRNAs and then exposed to the indicated concentrations of sotorasib for 72 hours. B, Immunoblot analysis of PTPRR as well as total and phosphorylated forms of AKT and ERK in H2122 cells transfected with siRNAs as in ( A ) and then incubated with or without sotorasib (1 μmol/L) for 4 hours. C, Cell viability assay for H2122AR30 cells stably transfected with control (GFP) or PTPRR expression plasmids and treated with the indicated concentrations of sotorasib for 72 hours. D, Immunoblot analysis of total or phosphorylated forms of EGFR, AKT, and ERK in H2122AR30 cells treated with sotorasib (1 μmol/L), cetuximab (10 μg/mL), or the combination of both drugs for 4 hours. E, Colony formation assay for H2122 and H2122AR30 cells incubated with or without sotorasib (1 μmol/L), cetuximab (10 μg/mL), or both drugs for 4 weeks. Surviving cells were stained with crystal violet. Data in A and C are means ± SD ( n = 3 independent experiments), and those in B , D , and E are representative of at least three independent experiments.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: Viability Assay, Transfection, Control, Western Blot, Incubation, Stable Transfection, Expressing, Colony Assay, Staining

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: Expression of PTPRR is regulated by promoter DNA methylation in KRAS G12C -mutant NSCLC cell lines. A, The human PTPRR gene including the promoter regions for the long and short isoforms of the encoded protein is shown. The positions of CpG islands as well as the histone marks H3K4me1, H3K4me3, and H3K27ac are indicated. B, Abundance of transcripts for the long and short isoforms of PTPRR in tumor specimens from patients with NSCLC in the dbGaP database ( n = 73). Data are presented as dot plots with means ± SD indicated by the dashed and solid lines for the short isoform, and they were analyzed with the Wilcoxon ranked-sum test. C, RT-qPCR analysis of mRNA abundance for the short form of PTPRR in the indicated KRAS G12C -mutant cell lines. Data are presented as dot plots, with means + SD being indicated by the dashed and solid lines ( n = 3 independent experiments) and analyzed by one-way ANOVA and Dunnett’s test. D, Bisulfite genomic sequencing of the promoter region for the short variant of PTPRR in the indicated KRAS G12C -mutant cell lines. The percentage of methylated CpG sites among total CpG sites is shown. Data are presented as violin plots, with the median and upper quartile values indicated by the dashed and dotted lines and analyzed by one-way ANOVA and Dunnett’s test. Data are representative of three independent experiments.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: Expressing, DNA Methylation Assay, Mutagenesis, Quantitative RT-PCR, Genomic Sequencing, Variant Assay, Methylation

Journal: Cancer Research Communications

Article Title: Constitutive EGFR Activation Induced by PTPRR Downregulation Confers Resistance to KRAS Inhibitors

doi: 10.1158/2767-9764.CRC-25-0489

Figure Lengend Snippet: Clinical relevance of PTPRR expression to sotorasib treatment in individuals with NSCLC. A, RT-qPCR analysis of PTPRR mRNA (short form) abundance in normal lung tissue ( n = 9) and NSCLC tumor tissue ( n = 10). Data are presented as dot plots with means ± SD indicated by the dashed and solid lines, and they were analyzed with the Wilcoxon ranked-sum test. B, Representative IHC images for specimens of normal lung tissue (negative) and NSCLC tumor tissue (low and high) stained with antibodies to PTPRR. Scale bars, 100 μm. C and D, Kaplan–Meier analysis of PFS ( C ) and OS ( D ) according to low ( n = 5) or high ( n = 8) status for cytoplasmic PTPRR staining in tumor cells for patients with KRAS G12C -mutant NSCLC treated with sotorasib.

Article Snippet: The KRAS G12C -mutant NSCLC cell lines NCI-H2122 (RRID: CVCL_1531), SW1573 (RRID: CVCL_1720), and NCI-H358 (RRID: CVCL_1559); the EGFR -mutated NSCLC cell line NCI-H1975 (RRID: CVCL_1511); and the KRAS G12C -mutant colorectal cancer cell line SW837 (RRID: CVCL_1729) were obtained from the ATCC.

Techniques: Expressing, Quantitative RT-PCR, Staining, Mutagenesis