Journal: Journal of Medicinal Chemistry

Article Title: RTx-303, an Orally Bioavailable Polθ Polymerase Inhibitor That Potentiates PARP Inhibitors in BRCA Mutant Tumors

doi: 10.1021/acs.jmedchem.5c00551

Figure Lengend Snippet: Cellular and in vitro activity of RTx-284. A. Bar plot showing the IC50 of RTx-284 in the indicated HR-proficient (gray) and HRD (red) cell lines. Data represent mean of at least 3 independent experiments performed in triplicate. B-D. Synergy plots created by Combenefit showing synergistic activity between RTx-284 and indicated PARPi in the indicated HRD cell lines. E. Bar plot showing that 10 μM RTx-284 suppresses the MMEJ GFP reporter. Data represent mean of 3 individual experiments performed in triplicate ±SEM. F. Dot plot showing increase of γH2AX following RTx-284:olaparib treatment in DLD1 BRCA2–/– cells. G. Bar plot showing that RTx-284 promotes an increase in caspase positive DLD1 BRCA2–/– cells. Data represent mean of 3 independent experiments performed in triplicate, ±SEM.

Article Snippet: HCT 116 BRCA2 −/– and HCT 116 Parental were obtained from Cancertools, London, UK.

Techniques: In Vitro, Activity Assay

Journal: Journal of Medicinal Chemistry

Article Title: RTx-303, an Orally Bioavailable Polθ Polymerase Inhibitor That Potentiates PARP Inhibitors in BRCA Mutant Tumors

doi: 10.1021/acs.jmedchem.5c00551

Figure Lengend Snippet: RTx-303 potentiates PARPi in BRCA mutant cells and xenografts. A. Scatter plot showing relative viability of the indicated ID8 cells treated with the indicated concentrations of olaparib with and without 5 μM RTx-303. Data represent mean of 3 independent experiments performed in triplicate, ±SEM. B. Scatter plot showing BRCA2 mutant BR-05–0566 PDX volumes following treatment with vehicle, 45 mg/kg olaparib (PO,QD), 60 mg/kg RTx-303 (PO,BID), and 45 mg/kg olaparib (PO,QD) with 60 mg/kg RTx-303 (PO,BID) (left). Scatter plot showing % body weight change (right). Data represent mean, n = 7 ±SEM. C. Scatter plot showing BRCA2 mutant BR-05–0568 PDX volumes following treatment with vehicle, 0.12 mg/kg talazoparib (PO,QD) with and without 60 mg/kg RTx-303 (PO,BID) (left). Scatter plot showing % body weight change. (right). Data represent mean, n = 6 ±SEM. D. Scatter plot showing MDA-MB-436 tumor volumes following treatment with vehicle, 45 mg/kg olaparib (PO,QD) with and without 60 mg/kg RTx-303 (PO,BID) (left). Scatter plot showing % body weight change (right). Data represent mean, n = 8 ±SEM. * p < 0.05, *** p < 0.001.

Article Snippet: HCT 116 BRCA2 −/– and HCT 116 Parental were obtained from Cancertools, London, UK.

Techniques: Mutagenesis

Journal: Breast Cancer : Targets and Therapy

Article Title: Identification of genes involved in breast cancer and breast cancer stem cells

doi: 10.2147/BCTT.S85202

Figure Lengend Snippet: Gene expression among different cancer types. Notes: Relative gene expression analysis of nine genes that were overexpressed in breast cancer and breast CSCs. The gene expression also was studied in colon and lung cancer, colon CSCs, lung CSCs, embryonic stem cells, and in a reference sample. The experiments were performed in triplicate and a P -value of <0.05 was considered significant. Abbreviations: CSCs, cancer stem cells; ESCs, embryonic stem cells; HP , haptoglobin; SMPD1 , sphingomyelin phosphodiesterase 1, acid lysosomal; SCRIB , scribbled homolog ( Drosophila ); KCMF1 , potassium channel modulatory factor 1; FAM155B , family with sequence similarity 155, member B; PTGER3 , prostaglandin E receptor 3 (subtype EP3); GPR3 , G protein-coupled receptor 3; TMX2 , thioredoxin-related transmembrane protein 2; DDX49 , DEAD (Asp-Glu-Ala-Asp) box polypeptide 49.

Article Snippet: During the exponential phase of proliferation, commercial breast CSCs cells were plated in 24-well plates (E36102-29-24Well; Celprogen) and transfected with gene-specific small interfering RNAs (siRNAs) using Lipofectamine 2000 Reagent (11668-027; Thermo Fisher Scientific), according to the manufacturer’s instructions.

Techniques: Expressing, Sequencing

Journal: Breast Cancer : Targets and Therapy

Article Title: Identification of genes involved in breast cancer and breast cancer stem cells

doi: 10.2147/BCTT.S85202

Figure Lengend Snippet: Breast CSCs pre- and post-siRNA knockdown. Note: Representative images showing breast CSCs pre- and post-siRNA knockdown. Abbreviations: CSCs, cancer stem cells; siRNA, small interfering RNA; TMX2 , thioredoxin-related transmembrane protein 2; FAM155B , family with sequence similarity 155, member B; PTGER3 , prostaglandin E receptor 3 (subtype EP3); GPR3 , G protein-coupled receptor 3; DDX49 , DEAD (Asp-Glu-Ala-Asp) box polypeptide 49.

Article Snippet: During the exponential phase of proliferation, commercial breast CSCs cells were plated in 24-well plates (E36102-29-24Well; Celprogen) and transfected with gene-specific small interfering RNAs (siRNAs) using Lipofectamine 2000 Reagent (11668-027; Thermo Fisher Scientific), according to the manufacturer’s instructions.

Techniques: Small Interfering RNA, Sequencing

Journal: Breast Cancer : Targets and Therapy

Article Title: Identification of genes involved in breast cancer and breast cancer stem cells

doi: 10.2147/BCTT.S85202

Figure Lengend Snippet: Gene expression of stemness transcription factors in breast CSCs. Notes: Relative gene expression of transcription factors in breast CSCs following knockdown. The ∆∆Ct method was used to perform the analysis. Each bar represents the average of the Ct values. The assays were performed in triplicate and a P -value of <0.05 was considered to be significant. The assays are presented in a log2 scale. Thus, positive values indicate overexpression while negative values indicate underexpression. Abbreviations: CSCs, cancer stem cells; TMX2 , thioredoxin-related transmembrane protein 2; FAM155B , family with sequence similarity 155, member B; PTGER3 , prostaglandin E receptor 3 (subtype EP3); GPR3 , G protein-coupled receptor 3; DDX49 , DEAD (Asp-Glu-Ala-Asp) box polypeptide 49; NANOG , Homeobox protein NANOG; OCT3/4 , Octamer-binding transcription factor ¾; SOX2 , sex determining region Y-box 2; CD34 , Hematopoietic progenitor cell antigen CD34.

Article Snippet: During the exponential phase of proliferation, commercial breast CSCs cells were plated in 24-well plates (E36102-29-24Well; Celprogen) and transfected with gene-specific small interfering RNAs (siRNAs) using Lipofectamine 2000 Reagent (11668-027; Thermo Fisher Scientific), according to the manufacturer’s instructions.

Techniques: Expressing, Over Expression, Sequencing, Binding Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Ailanthone synergizes with PARP1 inhibitor in tumour growth inhibition through crosstalk of DNA repair pathways in gastric cancer

doi: 10.1111/jcmm.18033

Figure Lengend Snippet: AIL with OLP blocks HSP90‐BRCA1 and PARP1 activity. The protein expression of HSP90, BRCA1, PARP1 in two GC cell lines after treated with a blank control group, OLP, AIL and OLP + AIL group. (B) The protein expression of HSP90, BRCA1 in two GC cell lines after being treated with a specified concentration of AIL for 24 h. (C) Pull‐down of selected proteins with HSP90 in the nucleus and cytoplasm of GC cells treated with AIL for 24 h, and then the IP fractions were immunoblotted with HSP90, BRCA1 and GAPDH. (D) Representative images of HSP90, BRCA1 and PARP1 in PDX tumour tissue as detected via immunohistochemistry. Scale bars, 200 μm. (E) Data were derived from experiments conducted in triplicate in (D). The PDX tumour tissues were compared with the control group, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BRCA1 (1:100, Proteintech, 22362‐1‐AP).

Techniques: Activity Assay, Expressing, Control, Concentration Assay, Immunohistochemistry, Derivative Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Ailanthone synergizes with PARP1 inhibitor in tumour growth inhibition through crosstalk of DNA repair pathways in gastric cancer

doi: 10.1111/jcmm.18033

Figure Lengend Snippet: AIL inhibits BRCA1 through downregulating P23 and inevitably suppress the HR pathway of GC. Western blotting was performed to detect the expression of P23, HSP90 and BRCA1 in AGS and SGC7901 cells after treatment with a specified concentration of CEL (P23 inhibitor) for 24 h. (B) The mRNA expression of P23 in two GC cell lines (AGS and SGC7901) after being treated with P23 shRNA lentivirus. (C) The protein expression of P23, HSP90 and BRCA1 in two GC cell lines after being treated with shP23‐1, shP23‐2 and shP23‐3. (D‐F) BALB/C nude mice are implanted with shNC or shP23‐3 GC cells subcutaneously. (D) The images of subcutaneous tumour nodules formed in two groups of mice. (E) Representative images of P23, HSP90 and BRCA1 in CDX tumour tissue as detected via immunohistochemistry. Scale bars, 50 μm. (F) Data were derived from experiments conducted in triplicate in (E).

Article Snippet: BRCA1 (1:100, Proteintech, 22362‐1‐AP).

Techniques: Western Blot, Expressing, Concentration Assay, shRNA, Immunohistochemistry, Derivative Assay