Journal: Cancers

Article Title: Multidrug-Resistant Profiles in Non-Small Cell Lung Carcinoma Patient-Derived Cells: Implications for Personalized Approaches with Tyrosine Kinase Inhibitors

doi: 10.3390/cancers16111984

Figure Lengend Snippet: Response of cancer cells in patient-derived NSCLC cultures to tyrosine kinase inhibitors. ( a ) The NSCLC cultures were categorized into six groups according to the response of the CK8/18+ cells to the TKIs, from “best responder” (strong response, IC50 closest to the lowest TKI concentration) to “non-responder”. ( b ) The intrinsic expression of ABC transporters ABCB1, ABCC1, and ABCG2 in NSCLC cultures. The graphs show the percentage of cells positive for the ABCB1, ABCC1, and ABCG2 antibodies. NSCLC cultures were considered inherently resistant if at least 20% of the cells were positive for at least one ABC transporter (red line). Data are presented as mean ± SEM ( n = 4).

Article Snippet: The cells were then incubated overnight at 4 °C with a primary rabbit CK8/18 antibody cocktail (clone SU0338, #MA5-32118, Thermo Fisher Scientific, Waltham, MA, USA) and primary mouse antibodies against ABCB1 (clone C219, #MA1-26528, Thermo Fisher Scientific, Waltham, MA, USA), ABCC1 (clone IU5C1, #MA5-16079, Thermo Fisher Scientific, Waltham, MA, USA), or ABCG2 (clone 1H2, #ab130244, Abcam, Cambridge, UK).

Techniques: Derivative Assay, Concentration Assay, Expressing

Journal: Cancers

Article Title: Multidrug-Resistant Profiles in Non-Small Cell Lung Carcinoma Patient-Derived Cells: Implications for Personalized Approaches with Tyrosine Kinase Inhibitors

doi: 10.3390/cancers16111984

Figure Lengend Snippet: NSCLC patient-derived cultures with increased ABCB1, ABCC1, and ABCG2 expression after treatment with tyrosine kinase inhibitors.

Article Snippet: The cells were then incubated overnight at 4 °C with a primary rabbit CK8/18 antibody cocktail (clone SU0338, #MA5-32118, Thermo Fisher Scientific, Waltham, MA, USA) and primary mouse antibodies against ABCB1 (clone C219, #MA1-26528, Thermo Fisher Scientific, Waltham, MA, USA), ABCC1 (clone IU5C1, #MA5-16079, Thermo Fisher Scientific, Waltham, MA, USA), or ABCG2 (clone 1H2, #ab130244, Abcam, Cambridge, UK).

Techniques: Expressing

Journal: Cancers

Article Title: Multidrug-Resistant Profiles in Non-Small Cell Lung Carcinoma Patient-Derived Cells: Implications for Personalized Approaches with Tyrosine Kinase Inhibitors

doi: 10.3390/cancers16111984

Figure Lengend Snippet: Patient-derived NSCLC cultures with increased expression of MDR markers after treatment with erlotinib. The CK8/18 antibody was used to differentiate between cancer cells and non-cancer cells in a mixed culture. The graphs show the percentage of cells positive for antibodies ABCB1 ( a ), ABCC1 ( b ) and ABCG2 ( c ) for each experimental condition. Data are presented as mean ± SEM ( n = 4). Statistically significant differences between the control and treated groups showing an increase of at least 20% in ABCB1, ABCC1 and ABCG2 expression are indicated by * ( p < 0.05).

Article Snippet: The cells were then incubated overnight at 4 °C with a primary rabbit CK8/18 antibody cocktail (clone SU0338, #MA5-32118, Thermo Fisher Scientific, Waltham, MA, USA) and primary mouse antibodies against ABCB1 (clone C219, #MA1-26528, Thermo Fisher Scientific, Waltham, MA, USA), ABCC1 (clone IU5C1, #MA5-16079, Thermo Fisher Scientific, Waltham, MA, USA), or ABCG2 (clone 1H2, #ab130244, Abcam, Cambridge, UK).

Techniques: Derivative Assay, Expressing, Control

Journal: Cancers

Article Title: Multidrug-Resistant Profiles in Non-Small Cell Lung Carcinoma Patient-Derived Cells: Implications for Personalized Approaches with Tyrosine Kinase Inhibitors

doi: 10.3390/cancers16111984

Figure Lengend Snippet: Patient-derived NSCLC cultures with an increased expression of ABCB1 after treatment with nintedanib. The CK8/18 antibody was used to differentiate between cancer cells and non-cancer cells in a mixed culture. The graphs show the percentage of cells positive for the ABCB1 antibody for each experimental condition. Data are presented as mean ± SEM ( n = 4). Statistically significant differences between the control and treated groups showing an increase in ABCB1 expression of at least 20% are indicated by * ( p < 0.05).

Article Snippet: The cells were then incubated overnight at 4 °C with a primary rabbit CK8/18 antibody cocktail (clone SU0338, #MA5-32118, Thermo Fisher Scientific, Waltham, MA, USA) and primary mouse antibodies against ABCB1 (clone C219, #MA1-26528, Thermo Fisher Scientific, Waltham, MA, USA), ABCC1 (clone IU5C1, #MA5-16079, Thermo Fisher Scientific, Waltham, MA, USA), or ABCG2 (clone 1H2, #ab130244, Abcam, Cambridge, UK).

Techniques: Derivative Assay, Expressing, Control

Journal: International journal of molecular sciences

Article Title: Talazoparib Does Not Interact with ABCB1 Transporter or Cytochrome P450s, but Modulates Multidrug Resistance Mediated by ABCC1 and ABCG2: An in Vitro and Ex Vivo Study.

doi: 10.3390/ijms232214338

Figure Lengend Snippet: Figure 2. MDCKII cell accumulation studies showing effects of talazoparib on transport activities of (A) ABCB1, (B) ABCG2, and (C) ABCC1. The cells were pre-exposed to talazoparib or model inhibitors (LY335979, Ko143, and MK571, respectively) and then treated with fluorescent chemotherapeutics, daunorubicin, or mitoxantrone. After the substrate accumulation period, the cells were trypsinized, and flow cytometer was used to quantify the intracellular fluorescence. The plotted values of relative accumulation reflect a fold increase in the accumulation of the probe substrate caused by the tested compound; they are numerically expressed as ratios of relative fluorescence units (RFUs) from treated samples to RFUs of vehicle control. Model inhibitors and vehicle controls represented 100% inhibition and 0% inhibition, respectively; their values were applied during normalization of talazoparib data and calculation of its IC50 values. Statistical analysis was accomplished using one- way ANOVA followed by Dunnett’s post hoc test (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 relative to control), while the presented data are means ± SD from three independent assays. DAU, daunorubicin; MTX, mitoxantrone; TAL, talazoparib.

Article Snippet: Primary antibodies against human ABCB1 (cat. no. sc-13131), ABCG2 (cat. no. sc-377176), ABCC1 (cat. no. sc-18835), and secondary antimouse antibody (cat. no. sc-516102) were bought from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Cytometry, Control, Inhibition

Journal: International journal of molecular sciences

Article Title: Talazoparib Does Not Interact with ABCB1 Transporter or Cytochrome P450s, but Modulates Multidrug Resistance Mediated by ABCC1 and ABCG2: An in Vitro and Ex Vivo Study.

doi: 10.3390/ijms232214338

Figure Lengend Snippet: Figure 9. The effect of talazoparib treatment on the ABCB1, ABCC1, and ABCG2 mRNA levels in breast cancer MCF-7 model. (A) The MTT viability assay performed after the 48 h incubation with a tested drug. (B) Gene induction studies. The MCF-7 cells were treated with talazoparib (0.5 µM) or rifampicin (25 µM). The qRT-PCR technique was employed for the assessment of the target genes’ mRNA expressions after 24 and 48 h incubation interval. The dotted lines draw the boundaries for downregulation (lower line) and upregulation (upper line) according to the EMA’s DDI-testing guidelines [24]. The plotted data are means ± SD from three independent repetitions. RIF, rifampicin; TAL, talazoparib.

Article Snippet: Primary antibodies against human ABCB1 (cat. no. sc-13131), ABCG2 (cat. no. sc-377176), ABCC1 (cat. no. sc-18835), and secondary antimouse antibody (cat. no. sc-516102) were bought from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: MTT Viability Assay, Incubation, Quantitative RT-PCR