Journal: International journal of cancer

Article Title: MED15, encoding a subunit of the mediator complex, is overexpressed at high frequency in castration-resistant prostate cancer.

doi: 10.1002/ijc.28647

Figure Lengend Snippet: Figure 5. MED15 knockdown and proliferation in PCa cells. Correlation of MED15 expression with tumor proliferation markers Ki67 and PHH3 (a). Expression of MED15 in PC3 (b) and DU145 (c) nuclear extracts by Western blot. SiRNA mediated MED15 knockdown in DU145 and PC3 cell lines by Western blot (d) and IHC (e). MTT proliferation assay of PC3 cells (f). *p < 0.05, **p < 0.01, ***p < 0.001. Similarly, proliferation assay of DU145 cells (g). *p < 0.05, **p < 0.01, ***p < 0.001. SiRNA mediated MED15 knockdown in LNCaP cell line (h). Prolif- eration assay of LNCaP cells treated with DHT (i). *p < 0.05, **p < 0.01, ***p < 0.001. ***p < 0.001, Pearson. ShRNA stable knockdown of MED15 in the DU145 cell line (j). Proliferation assay of DU145 cells with stable knockdown of MED15 compared to the scrambled control (k). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: Cell lysates and nuclear extracts of PC3 (sc-2152, Santa Cruz, CA) and DU145 (sc-24960, Santa Cruz, CA) were analyzed by immunobloting using MED15 rabbit polyclonal antibody (Proteintech, Chicago, IL).

Techniques: Knockdown, Expressing, Western Blot, Proliferation Assay, shRNA, Control

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Combined Treatment with Stattic and Docetaxel Alters the Bax/Bcl-2 Gene Expression Ratio in Human Prostate Cancer Cells

doi: 10.22034/APJCP.2016.17.11.5031

Figure Lengend Snippet: IC50 Values of Docetaxel and Stattic in DU145 Cells

Article Snippet: Human prostate cancer (DU145) cell line was obtained from National Cell Bank of Iran (Pasteur Institute, Iran).

Techniques: Incubation

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Combined Treatment with Stattic and Docetaxel Alters the Bax/Bcl-2 Gene Expression Ratio in Human Prostate Cancer Cells

doi: 10.22034/APJCP.2016.17.11.5031

Figure Lengend Snippet: DU145 Cells Exposed to Docetaxel and Stattic Treatments for 24, 48 and 72 h Determined by: MTT (A–C); Trypan Blue Cell Count (B-D). Data are Expressed as Cell Viability (%) ± SD of Three Independent Experiments.

Article Snippet: Human prostate cancer (DU145) cell line was obtained from National Cell Bank of Iran (Pasteur Institute, Iran).

Techniques: Cell Counting

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Combined Treatment with Stattic and Docetaxel Alters the Bax/Bcl-2 Gene Expression Ratio in Human Prostate Cancer Cells

doi: 10.22034/APJCP.2016.17.11.5031

Figure Lengend Snippet: Effect of Stattic, Docetaxel and Their Combination on the Expression of Bcl-2, Survivin, Mcl-1 and Bax in DU145 cells. A. The levels of Bcl-2, Survivin, Mcl-1 and Bax mRNA expression, followed by treatment with docetaxel (2 nM), stattic (2 µM) and their combination (stattic 1 µM+docetaxel 1 nM) for 24h, were determined using real-time PCR. Mean ± SD was calculated from three independent experiments(**p<0.01). B. Levels of Bcl-2, Mcl-1, Survivin and Bax expression were analyzed. Bands representing Bcl-2, Mcl-1, Survivin and Bax are shown.

Article Snippet: Human prostate cancer (DU145) cell line was obtained from National Cell Bank of Iran (Pasteur Institute, Iran).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Combined Treatment with Stattic and Docetaxel Alters the Bax/Bcl-2 Gene Expression Ratio in Human Prostate Cancer Cells

doi: 10.22034/APJCP.2016.17.11.5031

Figure Lengend Snippet: Effect of Stattic and/or Docetaxel on Bax/Bcl-2 Ratio. Bax/Bcl-2 Ratio was Calculated and Show That It was Significantly Increased in Stattic and/or Docetaxel Treated DU145 Cells (*p<0.05, **p<0.01).

Article Snippet: Human prostate cancer (DU145) cell line was obtained from National Cell Bank of Iran (Pasteur Institute, Iran).

Techniques:

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a SLFN11 expression in normal tissues, arrows indicate (from left to right); negative breast lobular cells, positive (red) and negative (blue) prostatic cells, positive lymphocytes, positive bronchial epithelial cells, negative colon epithelial cells. Negative Langerhans islets (blue circle) and positive cluster of acinar cells (red circle) also shown. b SLFN11 prevalence in various cancer types from multi-tumour tissue microarrays (TMAs), expression of SLFN11 shown as percent of positive tumour cells when internal control staining was acceptable. Median given (green) for each cancer type. c Example images of SLFN11 IHC staining in head and neck, thyroid, lymphoma and prostate cancers (Ca). d , e Breakdown of the SLFN11 protein levels by: d breast cancer subtype; oestrogen receptor-positive (ER + ), human epidermal growth factor receptor 2 (HER2 + ), triple-negative breast cancer (TNBC), invasive lobular breast cancer (ILC) and e thyroid cancer subtype; papillary and follicular. f – h SLFN11 expression in colorectal cancer (CRC) patients ( n = 144) subcategorised by clinical patient information: f presence of metastasis or not; g tumour grade, h disease stage. Data shown as median ± interquartile range. * P value <0.05 by Mann–Whitney test. Black arrows are stromal and endothelial cells. Scale bars at 100 µm.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Expressing, Control, Staining, Immunohistochemistry, MANN-WHITNEY

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a – c SLFN11 sub-clonal expression pattern demonstrated by IHC of a a CRC TMA core, b resected serous ovarian cancer tissue, c resected breast cancer tissue. A magnified image of the high- and low SLFN11 subclones are shown for each. Black arrows indicate stromal and endothelial cells used as a positive internal control. d RNA in situ hybridisation (ISH) of SLFN11 showing expression of SLFN11 RNA transcripts in the breast cancer SLFN11 subclones. e SLFN11 sub-clonal expression correlated to Ki67 expression. Line indicates patient-matched subclones. * P value < 0.05 paired t test. Scale bars at 50 µm unless otherwise stated.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Expressing, Control, In Situ, Hybridization

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a Image of a SCLC patient tumour demonstrating sub-clonal SLFN11 expression, areas from the high- and low-expressing subclones are magnified. The black arrow indicates SLFN11-positive stromal cells in the negative sub-clone. Scale bars at 100 µm unless otherwise stated. b SLFN11 prevalence by IHC in SCLC patients, median in green. c Bimodal prevalence of SLFN11 in the SCLC cohort ( n = 124). d SLFN11 expression in SCLC subcategorised by disease stage. Black; SLFN11 monoclonal expression, Red; SLFN11 sub-clonal tumours. Median ± interquartile range shown. e – h Kaplan–Meier analysis of SCLC patients categorised by SLFN11 expression; SLFN11 high (H-score >122) compared to low (H-score ≤122). e Progression-free survival (PFS) and f overall survival (OS) in months for patients ( n = 24) that received first-line platinum (carboplatin or cisplatin) plus etoposide. g PFS and h OS in months for 45 and 57 patients, respectively, that received any chemotherapy. Events table with patients at risk shown for each timepoint.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a Prevalence of SLFN11 by IHC H-score in serous ovarian cancer patient biopsies ( n = 151), cut-off depicting the high and low SLFN11 subgroups shown, b representative images of high and low SLFN11 tumours. Black arrow indicates SLFN11-positive stromal cells used as an internal control. Scale bars at 100 µm. c SLFN11 H-score of patients divided by extreme good and poor clinical responders to carboplatin and paclitaxel doublet therapy. Median ± interquartile range shown. d SLFN11 IHC protein expression by H-score compared to NanoString quantified SLFN11 gene expression in the HGSOC patients. Dotted line indicates gene expression threshold that distinguishes positive from negative SLFN11 patients. Patients with sub-clonal SLFN11 indicated by a red dot. e , f Kaplan–Meier survival curves of ( e ) progression-free interval (PFI) and ( f ) overall survival (OS) of high (>30 H-score SLFN11) vs. low SLFN11 ( ≤ 30 H-score SLFN11) subgroups in the 34 patients with high-grade serous ovarian cancer treated with carboplatin and paclitaxel doublet. Events table with patients at risk shown for each timepoint.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Control, Expressing, Gene Expression

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a Cox proportional hazards model of progression-free survival (PFS) of high (>30 H-score SLFN11) vs. low SLFN11 ( ≤ 30 H-score SLFN11) subgroups in platinum-sensitive serous ovarian carcinoma patients treated with either placebo or olaparib. b Overall survival (OS) of high vs. low SLFN11 in the same study. Events table with patients at risk shown for each timepoint.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques:

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a – d Cox proportional hazards model by SLFN11 high (>30 H-score SLFN11) vs. low SLFN11 ( ≤ 30 H-score SLFN11) subgroups in platinum-sensitive serous ovarian carcinoma patients treated with either placebo or olaparib for ( a ) progression-free survival (PFS) in BRCA wild-type patients, b PFS in BRCA mutant patients, c Overall survival (OS) in BRCA wild-type patients, d OS in BRCA mutant patients. Events table with patients at risk shown for each timepoint. VUS BRCAm patients included in the BRCAwt group.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Mutagenesis

Journal: Cancer Medicine

Article Title: Overexpression of claspin promotes docetaxel resistance and is associated with prostate‐specific antigen recurrence in prostate cancer

doi: 10.1002/cam4.4113

Figure Lengend Snippet: Effect of claspin inhibition on cell proliferation. (A) Western blot analysis of claspin in DU145 and PC3 cells transfected with CLSPN or negative control siRNAs. (B) qRT‐PCR analysis of CLSPN in DU145 and PC3 cells transfected with CLSPN or negative control siRNAs. (C) Effects of claspin knockdown on DU145 and PC3 cell proliferation. An MTT assay assessed cell proliferation at 1, 2, and 4 days after seeding on 96‐well plates. Bars and error bars indicate the mean and S.D., respectively

Article Snippet: DU145, PC3, and LNCaP were purchased from the Japanese Collection of Research Bioresources Cell Bank.

Techniques: Inhibition, Western Blot, Transfection, Negative Control, Quantitative RT-PCR, Knockdown, MTT Assay

Journal: Cancer Medicine

Article Title: Overexpression of claspin promotes docetaxel resistance and is associated with prostate‐specific antigen recurrence in prostate cancer

doi: 10.1002/cam4.4113

Figure Lengend Snippet: Association of claspin inhibition with DNA damage and resistance to docetaxel (DTX). (A) Dose‐dependent effect of DTX on the viability of DU145 and PC3 cells analyzed by MTT assay. Bars and error bars indicate the mean and S.D., respectively. (B) Western blot analysis of γ‐H2AX expressions in DU145 and PC3 cells transfected with CLSPN or negative control siRNAs after 24 h of treatment with DTX. (C) Western blot analysis of cleaved PARP expressions in DU145 and PC3 cells transfected with CLSPN or negative control siRNAs after 24 h of treatment with DTX

Article Snippet: DU145, PC3, and LNCaP were purchased from the Japanese Collection of Research Bioresources Cell Bank.

Techniques: Inhibition, MTT Assay, Western Blot, Transfection, Negative Control

Journal: Cancer Medicine

Article Title: Overexpression of claspin promotes docetaxel resistance and is associated with prostate‐specific antigen recurrence in prostate cancer

doi: 10.1002/cam4.4113

Figure Lengend Snippet: Effect of claspin inhibition on Akt, Erk, and ATR‐CHK1 pathways. (A) Western blot analysis of claspin, Akt, p‐Akt, Erk1/2, and p‐Erk1/2 in DU145 and PC3 cells transfected with CLSPN or negative control siRNAs. (B) Western blot analysis of claspin, CHK1, and p‐CHK1 in DU145 and PC3 cells transfected with CLSPN or negative control siRNAs

Article Snippet: DU145, PC3, and LNCaP were purchased from the Japanese Collection of Research Bioresources Cell Bank.

Techniques: Inhibition, Western Blot, Transfection, Negative Control

Journal: Cancer Medicine

Article Title: Overexpression of claspin promotes docetaxel resistance and is associated with prostate‐specific antigen recurrence in prostate cancer

doi: 10.1002/cam4.4113

Figure Lengend Snippet: Effect of claspin inhibition on the docetaxel (DTX)‐resistant DU145 (DU145‐DR) cell line. (A) Western blot analysis of claspin, Akt, p‐Akt, Erk1/2, and p‐Erk1/2 in DU145 and DU145‐DR cells transfected with CLSPN or negative control siRNAs. (B) Western blot analysis of claspin, CHK1, and p‐CHK1 in DU145‐DR cells transfected with CLSPN or negative control siRNAs. (C) Effects of claspin knockdown on DU145‐DR cell proliferation that was assessed by an MTT assay. (D) Dose‐dependent effect of DTX on the viability of DU145‐DR cells analyzed by MTT assay. Bars and error bars indicate the mean and S.D., respectively. (E) Western blot analysis of claspin, Akt, p‐Akt, ERK1/2, p‐ERK1/2, CHK1, and p‐CHK1 in DU145‐DR cells transfected with CLSPN or negative control siRNAs

Article Snippet: DU145, PC3, and LNCaP were purchased from the Japanese Collection of Research Bioresources Cell Bank.

Techniques: Inhibition, Western Blot, Transfection, Negative Control, Knockdown, MTT Assay