To identify epigenetically silenced genes in multiple myeloma (MM) cell lines and to determine the effects of 5-aza-2-deoxycytidine and trichostatin A on gene expression.  We treated 3 multiple myeloma cell lines (MM1, NCI-H929, U266) with 5-aza-2-deoxycytidine and/or trichostatin A.

Transcription profiling of three human multiple myeloma cell lines treated with 5-aza-2-deoxycytidine and/or trichostatin A

Genome-wide transcriptional response to 5-aza-2-deoxycytidine and trichostatin A in multiple myeloma cells

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

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Transcription profiling by array of three human multiple myeloma cell lines treated with 5-aza-2-deoxycytidine and/or trichostatin A

To identify epigenetically silenced genes in multiple myeloma (MM) cell lines and to determine the effects of 5-aza-2-deoxycytidine and trichostatin A on gene expression.  We treated 3 multiple myeloma cell lines (MM1, NCI-H929, U266) with 5-aza-2-deoxycytidine and/or trichostatin A.


P-AFFY-2

Labeling of samples was carried out according to the manufacturer’s standard protocol

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

RPMI 1640 supplemented with 10% FCS under humidified conditions.

5-aza-2-deoxycytidine: 0.5 µM Aza-dC for 7 days   Trichostatin A (TSA): 100 ng/ml TSA for 24 h   combination: 0.5 µM Aza-dC for 7 days and 100 ng/ml TSA for additional 24 h.   Control cells received no drug treatment.

Total cellular RNA was isolated from cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer´s instruction. 10µg of purified RNA were used for double-stranded cDNA synthesis. First- and second-strand cDNA synthesis was performed using the SuperScript double-stranded cDNA synthesis kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol with the use of an oligo-dT primer containing a T7 RNA polymerase promoter site. IVT was performed using the BioArray High Yield RNA Transcript Labeling kit (Enzo Life Sciences, Farmingdale, US). 15µg of cRNA were fragmented at 94°C for 35 minutes in a fragmentation buffer (Affymetrix, Santa Clara, CA).

Transcription profiling by array of three human multiple myeloma cell lines treated with 5-aza-2-deoxycytidine and/or trichostatin A

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