Affymetrix GeneChip Human Genome HG-U133A [HG-U133A]

Delineation of molecular mechanisms of sensitivity to lapatinib in breast cancer cell lines using global gene expression profiles

rawData

scan

processedDataMatrix

Dose and time course response of lapatinib in breast cancer cell lines.

cell line

compound

dose

time

Transcription profiling by array of human breast cancer cell lines after treatment with lapatinib

Transcription profiling of lapatinib treated of breast cancer cell lines

Dose and time course response of lapatinib in breast cancer cell lines.


Affymetrix:Protocol:ExpressionStat

Title: Affymetrix CHP Analysis (ExpressionStat). Description:

Affymetrix:Protocol:Hybridization-EukGE-WS2v4_450

Title: Affymetrix EukGE-WS2v4_450 Hybridization. Description:

Affymetrix:Protocol:Hybridization-Unknown

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MEXP-12318

MDA-MB-468, SK-Br-3, T-47D and BT-474 breast tumor cell lines were obtained from the American Type Culture Collection (Manassas, VA).  MDA-MB-468 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, Invitrogen, Carlsbad, CA, catalog number 12430-054) containing 10% Fetal Bovine Serum (FBS, HyClone, Logan, UT, catalog number SH30071.03).  SK-Br-3, T-47D and BT-474 were cultured in RPMI 1640 (RPMI, Invitrogen, catalog number 22400-089) containing 10% FBS.  Cells were plated in 100 mm tissue culture dishes (Becton Dickinson, Franklin Lakes, NJ, catalog number 35-3003) and allowed to grow until cells were approximately 70% confluent.  All cells were cultured in a humidified incubator at 37 C in 95% air/5% CO2.

P-MEXP-12319

Powdered GW572016F was synthesized as described (Carter, MC, et al.  1999 PCT Appl.) and dissolved in neat dimethyl sulfoxide (DMSO, Sigma Aldrich Chemical Company, St. Louis, MO, catalog number D2650) at ten millimolar.  The ten millimolar stock was serially diluted in neat DMSO to make stocks of one millimolar and 0.1 millimolar.  The serially diluted DMSO stocks of GW572016F were added to the appropriate growth media for each cell line at a 1:1000 dilution to make treatment solutions of one micromolar and 0.1 micromolar in growth media.  These solutions contained 0.1% DMSO.  A vehicle control containing 0.1 % DMSO was also made.  The media from the cell plates were aspirated and ten milliliters media containing vehicle, one micromolar or 0.1 micromolar GW572016F were added to each of three plates.  One set of three plates were untreated to be used as a negative control.  Cells were returned to the incubator and treated for the times described in the data tables.  After appropriate incubation times, the media were aspirated and three milliliters of TRIzol Reagent (Invitrogen, catalog number 15596-018) were added to each plate.  The plates were scraped to ensure removal of all cells.  The lysate was collected into 15 milliliter polypropylene tubes  and transferred to a -80 C freezer.

P-MEXP-12320

Cells were lysed in TRIzol®, the cell lysate was extracted with chloroform, and RNA was purified from the lysate using the QIAGEN RNeasy® Mini Kit (QIAgen Inc., Valencia, CA.).  cDNA was prepared from 5 ug total RNA using the Invitrogen™ SuperScript™ Double-Stranded cDNA Synthesis Kit (Invitrogen Inc, Carlsbad, CA). cDNA was amplified and biotinylated using the ENZO BioArray™ HighYield™ RNA Transcript Labeling Kit (Enzo Biochem, Inc., Farmingdale, NY).  The amplified cDNA samples were fragmented and hybridized onto HG-U133A GeneChips®. After sample hybridization, arrays were washed, stained and scanned according to the standard Affymetrix® protocol. 

P-MEXP-12334

 Cells plated in triplicate were lysed in TRIzol®, the cell lysate was extracted with chloroform, and RNA was purified from the lysate using the QIAGEN RNeasy® Mini Kit (QIAgen Inc., Valencia, CA.).  cDNA was prepared from 5 ug total RNA using the Invitrogen™ SuperScript™ Double-Stranded cDNA Synthesis Kit (Invitrogen Inc, Carlsbad, CA). cDNA was amplified and biotinylated using the ENZO BioArray™ HighYield™ RNA Transcript Labeling Kit (Enzo Biochem, Inc., Farmingdale, NY).  The amplified cDNA samples were fragmented and hybridized onto HG-U133A GeneChips®. After sample hybridization, arrays were washed, stained and scanned according to the standard Affymetrix® protocol.  

Transcription profiling by array of human breast cancer cell lines after treatment with lapatinib

Submitter Supplied Information

Samples