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BioAssay: AID 624296

A quantitative high throughput screen for small molecules that induce DNA re-replication in MCF 10a normal breast cells.

Eukaryotic cells normally restrict genome duplication to once per cell division. When this process is aberrant, DNA re-replication occurs when cells reinitiate DNA replication before the first round of replication is complete. DNA re-replication induces the DNA damage response, arrests cell proliferation, and triggers apoptosis. The cell cycle components cyclin A, Cdt-1, Cdc6 and geminin are more ..
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 Tested Compounds
 Tested Compounds
All(343222)
 
 
Active(9886)
 
 
Inactive(282614)
 
 
Inconclusive(51153)
 
 
 Tested Substances
 Tested Substances
All(345034)
 
 
Active(9908)
 
 
Inactive(283849)
 
 
Inconclusive(51277)
 
 
AID: 624296
Data Source: NCGC (DNARRMCF10aPRI)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-06-15

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 9886
Related Experiments
AIDNameTypeComment
463113qHTS assay for small molecules that induce DNA re-replication: SummarySummarydepositor-specified cross reference
463097Validation screen for small molecules that induce DNA re-replication in MCF 10A normal breast cellsConfirmatorysame project related to Summary assay
493165Retest of LOPAC molecules that induce DNA re-replication in SW480 adenocarcinoma cells.Confirmatorysame project related to Summary assay
493166Retest of LOPAC molecules that induce DNA re-replication in MCF 10A normal breast cells.Confirmatorysame project related to Summary assay
504364Validation screen for small molecules that induce DNA re-replication in SW480 colon adenocarcinoma cellsConfirmatorysame project related to Summary assay
504806Followup screen of LOPAC molecules that induce DNA re-replication in SW480 adenocarcinoma cellsConfirmatorysame project related to Summary assay
504809Followup screen for small molecules that induce DNA re-replication in MCF 10A normal breast cellsConfirmatorysame project related to Summary assay
624297A quantitative high throughput screen for small molecules that induce DNA re-replication in SW480 colon adenocarcinoma cells.Confirmatorysame project related to Summary assay
Description:
H Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]

MLPCN Grant: Fast Track K99 CA136555
Assay Provider: Melvin DePamphilis, NICHD, NIH

Assay Overview

Eukaryotic cells normally restrict genome duplication to once per cell division. When this process is aberrant, DNA re-replication occurs when cells reinitiate DNA replication before the first round of replication is complete. DNA re-replication induces the DNA damage response, arrests cell proliferation, and triggers apoptosis. The cell cycle components cyclin A, Cdt-1, Cdc6 and geminin are critical regulators of this process and their misregulation results in DNA re-replication in normal and cancer cells. To identify compounds that induce re-replication in normal cells, an image-based assay was used to measure DNA content of nuclei, as detected by the fluorescent DNA binding dye, Hoechst. Cell nuclei were imaged by a fluorescence scanning plate imager and compounds that induce DNA re-replication were identified by the increase of nuclei with elevated fluorescence intensity
Protocol
For screening, 250 cells in 5 uL/well were dispensed into black clear 1536-well plates (Aurora) using a solenoid-based dispenser. Following transfer of 23 nL compound or DMSO vehicle by a pin tool, the plates were incubated 48 hr at 37 C and 5% CO2. After addition of 1 uL/well Hoescht stain, the plates were incubated 40 min at 37 C, 5% CO2 and total intensity of fluorescent objects (nuclei) were enumerated by an Acumen Explorer Ex3 (TTP LabTech) fluorescence scanning plate imager (405 nm excitation, 500-530 nm emission).
Keywords: NIH Roadmap, MLPCN, MLI, MLSMR, qHTS, NCGC, DNA replication, cancer
Comment
Compound Ranking:
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent activators are ranked higher than compounds that showed apparent inhibition.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
From ChEMBL:
Assay Type: Functional
Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Activity_ScoreActivity score.Integer
6Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0000590000 uM (5.9e-05μM**)% Activity at given concentration.Float%
16Activity at 0.0001320000 uM (0.000132μM**)% Activity at given concentration.Float%
17Activity at 0.0002739534 uM (0.000273953μM**)% Activity at given concentration.Float%
18Activity at 0.0006031963 uM (0.000603196μM**)% Activity at given concentration.Float%
19Activity at 0.00135 uM (0.00134872μM**)% Activity at given concentration.Float%
20Activity at 0.00295 uM (0.00295108μM**)% Activity at given concentration.Float%
21Activity at 0.00674 uM (0.0067391μM**)% Activity at given concentration.Float%
22Activity at 0.015 uM (0.0149446μM**)% Activity at given concentration.Float%
23Activity at 0.034 uM (0.0337455μM**)% Activity at given concentration.Float%
24Activity at 0.074 uM (0.0737298μM**)% Activity at given concentration.Float%
25Activity at 0.169 uM (0.168636μM**)% Activity at given concentration.Float%
26Activity at 0.369 uM (0.369124μM**)% Activity at given concentration.Float%
27Activity at 0.843 uM (0.842729μM**)% Activity at given concentration.Float%
28Activity at 1.841 uM (1.84066μM**)% Activity at given concentration.Float%
29Activity at 4.214 uM (4.21365μM**)% Activity at given concentration.Float%
30Activity at 9.220 uM (9.21994μM**)% Activity at given concentration.Float%
31Activity at 20.60 uM (20.6μM**)% Activity at given concentration.Float%
32Activity at 46.10 uM (46.1μM**)% Activity at given concentration.Float%
33Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: K99 CA136555

Data Table (Concise)
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