DOI: 10.5061/dryad.3bk3j9kxp
title: Data from: Investigating the combined effects of jadomycin B and celecoxib against triple-negative breast cancer using zebrafish larval xenografts

description: Breast cancer affects 1 in 8 Canadian women over their lifetime.
 Triple-negative breast cancer (TNBC) represents 10-20 % of all advanced
 stage breast cancers, often developing multi-drug resistance (MDR),
 commonly resulting in treatment failure. Jadomycin B (JB), a natural
 product of Streptomyces venezuelae, maintains cytotoxicity against MDR
 TNBCs, shown to be enhanced when combined with selective COX-2 inhibitor,
 celecoxib (CXB). Our objectives were to generate a fluorescent human TNBC
 cell line, as well as evaluate the toxicity and anticancer effect of JB
 combined with CXB using zebrafish larval xenografts. Fluorescent human
 TNBC MDA-MB-231 cells (231-EGFP) were generated and characterized for
 zebrafish larval xenografts. A maximum tolerated dose (MTD) in zebrafish
 larvae was determined for JB (20 mM) and CXB (5 mM). Zebrafish embryos
 were xenotransplanted with 231-EGFP cells and treated with the MTD of JB
 CXB. The combination of JB and CXB resulted in a 75% reduction in 231-EGFP
 fluorescence intensity, significantly higher than reductions caused by
 either drug alone (39% for JB, 15 % for CXB) (p &lt; 0.05). This study
 demonstrates the safety and efficacy of JB combined with CXB in a
 zebrafish larval xenograft model for human TNBC, enhancing anticancer
 effects, similar to effects previously determined in vitro.
description: # Data from: Investigating the combined effects of jadomycin B and
 celecoxib against triple-negative breast cancer using zebrafish larval
 xenografts Hailey M. Stack, Michael G. Morash, Brendan T. McKeown, Lee D.
 Ellis, Kerry B. Goralski Contact of Authors:  Hailey M. Stack Department
 of Pharmacology, Faculty of Medicine, Dalhousie University Halifax, NS,
 B3H 4R2 [hailey.stack@dal.ca](mailto:hailey.stack@dal.ca) 
 
 Kerry B. Goralski College of Pharmacy, Faculty of Health, Department of
 Pharmacology Halifax, NS B3H 4R2
 [kerry.goralski@dal.ca](mailto:kerry.goralski@dal.ca) 
 
 Dataset DOI:
 [10.5061/dryad.3bk3j9kxp](https://doi.org/10.5061/dryad.3bk3j9kxp) ##
 Description of the data and file structure raw_toxicity_data: excel file
 containing raw data collected from a phenotypic (fish embryo toxicity
 assay/FET) and a behavioural assay of various treatment groups (jadomycin
 B: 2.5-80 uM; celecoxib: 2.5-25 uM; combination treatment: jadomycin B
 (5-20 uM) + celecoxib (5-10 uM)). Phenotypic data is presented in percent
 survival (%) which was recorded every 24 hours over a 72 hour
 treatment. Phenotypic data is presented in difference in distance traveled
 during light-dark (mm) which was recorded during 30 mins of alternating
 light to dark transitions.  raw_data_MTT: excel file containing raw data
 collected from MTT metabolic activity assays of various treatments
 (jadomycin B (0.04-10 uM) + celecoxib (5uM) in
 MDA-MB-231-control, MDA-MB-231-mCherry, and MDA-MB-231-EGFP cell lines. 5
 technical replicates (R1-5) and 3 biological replicates (Rep1-3) were
 preformed for each treatment in each cell line. Raw data is presented in
 absorbance values as well as a calculated percent viability (viability)
 and standard deviation (stdev).  raw_injection_data: excel file containing
 raw data collected from zebrafish embryos xenografts treated with
 jadomycin B (20uM) +/- celecoxib (5uM) at 0 hours and 72 hours post
 treatment. Data is presented as sum intensity (ROI Sum Intensity)
 calculated from fluorescent (GFP channel) microscope images take with a
 Nikon AZ100 Multizoom Microscope (set at 10X magnification and 4X
 objective) of xenografts in zebrafish embryos. Intensity fold change was
 calculated from dividing 72 hours post treatment by 0 hours post treatment
 (72h/0h) for each individual embryo. All intensity fold change was
 normalized to the average intensity fold change of the vehicle control to
 calculate precent reduction.  Sequencing Files: .seq files (containing
 sequencing reads) from Sanger Sequencing analysis of fluorescently
 transfected and control cell lines. The .seq files are plain text (fasta)
 file of the final base sequence from the sequencer. List:
 HS337_A01_1A-187F.seq, HS337_A02_1A-808R.seq, HS337_A03_1B-187F.seq,
 HS337_A04_1B-808R.seq, HS337_A05_1C-187F.seq, HS337_A06_1C-808R.seq,
 HS337_A07_2A-183F.seq, HS337_A08_2A-803R.seq, HS337_A09_2B-183F.seq,
 HS337_A10_2B-803R.seq, HS337_A11_2C-183F.seq, HS337_A12_2C-803R.seq,
 HS337_B01_3G-187F.seq, HS337_B02_3G-182R.seq, HS337_B03_3R-187F.seq,
 HS337_B04_3R-182R.seq, HS337_B05_4G-804F.seq, HS337_B06_4G-803R.seq,
 HS337_B07_4R-804F.seq, HS337_B08_4R-803R.seq  ### Files and variables ####
 File: raw_toxicity_data_.xlsx **Description:** raw data collected from a
 phenotypic (fish embryo toxicity assay/FET) and a behavioural assay of
 various treatment groups (jadomycin B: 2.5-80 uM; celecoxib: 2.5-25 uM;
 combination treatment: jadomycin B (5-20 uM) + celecoxib (5-10 uM)).
 Phenotypic data was recorded manually from visual scoring and is presented
 in percent survival (%), which was recorded every 24 hours over a 72 hour
 treatment. Behavioural data was recorded using a Noldus Daniovision
 Behavioral Tracking instrument and is presented in difference in distance
 traveled during light-dark (mm) which was recorded during 30 mins of
 alternating light to dark transitions.  Any cells including
 "N/A" indicates a read that was excluded to phenotypic toxicity
 including major phenotypic toxicity/changes or death of the embryo. 
 
 #### File: raw_data_MTT.xlsx **Description:** raw data collected from MTT
 metabolic activity assays of various treatments (jadomycin B (0.04-10 uM)
 + celecoxib (5uM) in MDA-MB-231-control, MDA-MB-231-mCherry, and
 MDA-MB-231-EGFP cell lines. 5 technical replicates (R1-5) and 3 biological
 replicates (Rep1-3) were preformed for each treatment in each cell line.
 Raw data is presented in absorbance values (measured at 550 nm using a
 BioTek Synergy HT plate reader Gen5 v2.01 software). Calculated percent
 viability (%viability) and standard deviation (stdev) is also presented in
 this data. Cells including "N/A" for the Blank absorbance
 reading is due to only 3 blanks being used per plate compared to 5
 technical replicates used for each treatment per plate.  ##### #### File:
 raw_injection_data_.xlsx **Description:** contains raw data collected from
 zebrafish embryos xenografts treated with jadomycin B (20uM) +/- celecoxib
 (5uM) at 0 hours and 72 hours post treatment. Data is presented as sum
 intensity (ROI Sum Intensity) calculated from fluorescent (GFP channel)
 microscope images take with a Nikon AZ100 Multizoom Microscope (set at 10X
 magnification and 4X objective) of xenografts in zebrafish embryos.
 Intensity fold change was calculated from dividing 72 hours post treatment
 by 0 hours post treatment (72h/0h) for each individual embryo. All
 intensity fold change was normalized to the average intensity fold change
 of the vehicle control to calculate precent reduction.  #### #### File:
 (HS337_A01_1A-187F.seq, HS337_A02_1A-808R.seq, HS337_A03_1B-187F.seq,
 HS337_A04_1B-808R.seq, HS337_A05_1C-187F.seq, HS337_A06_1C-808R.seq,
 HS337_A07_2A-183F.seq, HS337_A08_2A-803R.seq, HS337_A09_2B-183F.seq,
 HS337_A10_2B-803R.seq, HS337_A11_2C-183F.seq, HS337_A12_2C-803R.seq,
 HS337_B01_3G-187F.seq, HS337_B02_3G-182R.seq, HS337_B03_3R-187F.seq,
 HS337_B04_3R-182R.seq, HS337_B05_4G-804F.seq, HS337_B06_4G-803R.seq,
 HS337_B07_4R-804F.seq, HS337_B08_4R-803R.seq) **Description:** All raw
 sequencing files (.seq) containing sequencing reads from Sanger Sequencing
 analysis of fluorescently transfected and control cell line to validate
 successful transfection.  For analysis in the publication, consensus
 amplicons were generated by aligning both forward and reverse strand
 sequencing reads (.seq files) as a megamer and exporting a consensus
 sequence using DNASTAR Laser gene core suite 12 analysis package (DNASTAR,
 USA). Sequences were aligned with the plasmid sequence (Addgene,
 Watertown, MA, USA), and the human genome sequence (GRCh38.p14 build,
 NCBI).