Journal: BBA Advances

Article Title: CEMIP promotes gefitinib resistance in lung cancer with EGFR-mutation possibly through forming a stable complex CEMIP/c-MYC

doi: 10.1016/j.bbadva.2025.100176

Figure Lengend Snippet: Characterization of gefitinib-resistant PC9GR cells and the role of CEMIP in gefitinib resistance. A, The scheme of induction of gefitinib resistance from PC9 cells. B, Cell viability of PC9 and PC9GR after treating gefitinib in 24 h, 48 h and 72 h. IC 50 values and RI of gefitinib (PC9GR/PC9) as shown below respectively. The resistant indexs of 24 h, 48 h and 72 h were 224.47, 677.39 and 952.14 respectively. C, PI/DAPI staining to assess cell deaths in PC9 and PC9GR cells with (lower panel) or without (upper panel) gefitinib treatment for 24 h, concentration of gefitinib was 0.1 μM for PC9, and 10 μM for PC9GR respectively. Magnification 100 × . D, FCM assay of cell apoptosis and viability in PC9 and PC9GR cells when treated with gefitinib for 48 h, concentration of gefitinib was 0.02 μM for PC9, and 10 μM for PC9GR respectively. E, The expression of p-EGFR and EGFR in PC9 and PC9GR cells under the treatment of Gefitinib (0.02 μM vs 10 μM) of 48 h. F, The binding effects of gefitinib to EGFR in PC9 and PC9GR cells, which were detected by CETSA assays. G, The expression of CEMIP in PC9 and PC9GR cells. H, The corelationship analysis between IC 50 value of gefitinib and the expression of CEMIP in the lung cancer cell lines, data were acquired from GDSC2 database. Results of Pearson correlation analysis were indicated in the upper panel of each chart, respectively. I, The validation of mRNA expression of CEMIP in PC9 cells with over-expressing CEMIP or PC9GR cells with interference of CEMIP expression. J, The validation of protein expression of CEMIP after transfecting with LV-encapsulated overexpressing or interference of CEMIP. K, Cell viability assays to determine gefitinib IC 50 in PC9 and PC9GR cells after transfecting with LV-encapsulated overexpressing or interference of CEMIP. Results ( n = 3) displayed as Mean ± SD, *, ** and #, ## presented P value < 0.05 and 0.01, respectively.

Article Snippet: EGFR Monoclonal antibody(MsIgG) , ProteinTech Co. (66455–1-Ig).

Techniques: Staining, Concentration Assay, Expressing, Binding Assay, Biomarker Discovery

Journal: bioRxiv

Article Title: Adaptive regulation of dNTP homeostasis confers osimertinib resistance in EGFR mutant non-small cell lung carcinoma

doi: 10.64898/2025.12.24.696437

Figure Lengend Snippet: (A) Chromatin immunoprecipitation followed by qPCR (ChIP-qPCR) was performed using an anti-RNAPII (8WG16) antibody to evaluate recruitment of RNA polymerase II to the RRM2 promoter, with or without osimertinib (Osi) treatment. Relative enrichment was normalized to the ACTB promoter. (B) ChIP-qPCR using HA-tagged MYBL2-overexpressing cell lines was conducted to assess MYBL2 binding to the RRM2 promoter. Promoter occupancy is shown relative to input. Data are presented as mean ± s.e.m. (n = 3). ***p < 0.001. (C) RRM2 mRNA expression was quantified by RT-qPCR in PC-9 and HCC827 cells following EGFR knockdown using siRNA. Transcript levels were normalized to GAPDH. (D) The impact of EGFR knockdown on MYBL2 recruitment to the RRM2 promoter was evaluated by ChIP-qPCR in HA-MYBL2-expressing cells transfected with control or EGFR siRNA. Promoter enrichment was normalized to input. Data are shown as mean ± s.e.m. (n = 3). ***p < 0.001. All data are representative of at least three independent experiments.

Article Snippet: RRM1 (#10526-1-AP), RRM2 (11661-1-AP), RRM2B (#18005-1-AP), CHK1 (#25887-1-AP), CHK2 (#13954-1-AP), EGFR (#66455-1-Ig), beta Actin (#20536-1-AP), HA tag (#51064-2-AP), Histone H3 (#17168-1-AP), GAPDH (#60004-1-Ig), POLD1(#15646-1-AP), POLH (#28133-1-AP), MYBL2 (#18896-1-AP) and TNNT3 (#19729-1-AP) were purchased from Proteintech.

Techniques: Chromatin Immunoprecipitation, ChIP-qPCR, Binding Assay, Expressing, Quantitative RT-PCR, Knockdown, Transfection, Control