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

Molecular effectors and modulators of hypericin-mediated cell death in bladder cancer cells

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Study of the gene expression of T24 bladder cancer cells in response to hypericin-mediated photodynamic therapy in the absence or presence of the p38 MAPK inhibitor PD169316

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Transcription profiling by array of human T24 bladder cancer cells in response to hypericin-mediated photodynamic therapy in the absence or presence of the p38 MAPK inhibitor PD169316

Study of the gene expression of T24 bladder cancer cells in response to hypericin-mediated photodynamic therapy in the absence or presence of the p38 MAPK inhibitor PD169316


Affymetrix:Protocol:Hybridization-EukGE-WS2v3

Title: Affymetrix EukGE-WS2v3 Hybridization. Description:

P-AFFY-2

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

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MEXP-86170

T24 cells (human transitional cell carcinoma of the bladder, obtained from American Type Culture Collection (ATCC)) were grown in Dulbecco’s Modified Eagle Medium (DMEM) containing L-glutamine (2 mM), penicillin (100 IU/ml), streptomycin (100 µg/ml), and 10 % foetal calf serum.  

P-MEXP-86172

For each condition, total RNA was isolated using the RNeasy Mini kit (Qiagen, Venlo, Netherlands) from 2 independent experiments performed in duplicate, generating 4 hybridizations per condition. RNA quality and concentration was determined using Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA). 

P-MEXP-86174

P-MEXP-86178

T24 cells were incubated with 150 nM hypericin for 16 h in subdued light conditions (< 1 µW/cm²). 

P-MEXP-86179

T24 cells were pre-incubated with 150 nM hypericin for 16 h in subdued light conditions (< 1 µW/cm²) and then 1uM of PD169316 was added in DMEM for 1h.

P-MEXP-86180

T24 cells were pre-incubated with 150 nM hypericin for 16 h in subdued light conditions (< 1 µW/cm²), followed by irradiation (4 J/cm²) for 7h.  Irradiation protocol: Cells were irradiated by placing the cell culture dishes on a plastic diffuser sheet 5 cm above a set of 7 L18W/30 fluorescent lamps (Osram, Berlin, Germany). The maximal emission of the fluorescent lamps was between 530 and 620 nm, which coincides with the absorption peak of hypericin (595 nm). At the surface of the diffuser, the uniform fluence rate was 4.5 mW/cm², as measured by an IL 1400 radiometer (International Light, Newbyport, MA, USA). The fluence or light dose (J/cm²) was calculated by multiplying the fluence rate with the time of irradiation. During the irradiation procedure, the temperature never exceeded 37°C.  

P-MEXP-86181

T24 cells were pre-incubated with 150 nM hypericin for 16 h in subdued light conditions (< 1 µW/cm²), followed by addition of 1uM of PD169316 in DMEM for 1h. cells were then irradiated (4 J/cm²) for 7h.   Irradiation protocol: Cells were irradiated by placing the cell culture dishes on a plastic diffuser sheet 5 cm above a set of 7 L18W/30 fluorescent lamps (Osram, Berlin, Germany). The maximal emission of the fluorescent lamps was between 530 and 620 nm, which coincides with the absorption peak of hypericin (595 nm). At the surface of the diffuser, the uniform fluence rate was 4.5 mW/cm², as measured by an IL 1400 radiometer (International Light, Newbyport, MA, USA). The fluence or light dose (J/cm²) was calculated by multiplying the fluence rate with the time of irradiation. During the irradiation procedure, the temperature never exceeded 37°C.  

Transcription profiling by array of human T24 bladder cancer cells in response to hypericin-mediated photodynamic therapy in the absence or presence of the p38 MAPK inhibitor PD169316

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