Cell Proliferation Assay with Targeted Therapies

The water-soluble tetrazolium (WST-1) proliferation assay (Roche Diagnostics) was used to measure cell proliferation after knockdown or stimulation as described earlier [17 (link)]. Cells were treated with cetuximab (1/10 µg/ml, Merck), trastuzumab (5/20 µg/ml, Roche), afatinib (0.5 µM, Biozol), DMSO (0.05%, afatinib solvent), trastuzumab solvent (described in [17 (link)]) or cetuximab solvent (8.48 mg/ml NaCl, 1.88 mg/ml Na₂HPO₄ × 7H₂O, 0.41 mg/ml NaH₂PO₄xH₂O) for 72 h (cell numbers see Table S1, Additional file 1). In case of stimulation, cells were treated with 5 ng/ml recombinant HBEGF or 15 ng/ml recombinant AREG (R&D Systems).

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Ebert K., Haffner I., Zwingenberger G., Keller S., Raimúndez E., Geffers R., Wirtz R., Barbaria E., Hollerieth V., Arnold R., Walch A., Hasenauer J., Maier D., Lordick F, & Luber B. (2022). Combining gene expression analysis of gastric cancer cell lines and tumor specimens to identify biomarkers for anti-HER therapies—the role of HAS2, SHB and HBEGF. BMC Cancer, 22, 254.

Publication 2022

WST-1) proliferation assay cetuximab trastuzumab afatinib recombinant HBEGF recombinant AREG

Afatinib Areg Assay Cell proliferation Cells Cetuximab Diagnostics Dmso Hbegf Nacl Solvent Tetrazolium Trastuzumab

Corresponding Organization :

Other organizations : Klinikum rechts der Isar, Technical University of Munich, University of Bonn, Helmholtz Centre for Infection Research, Helmholtz Zentrum München, Biomax Informatics (Germany)

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Variable analysis

independent variables

Cetuximab (1/10 µg/ml, Merck)
Trastuzumab (5/20 µg/ml, Roche)
Afatinib (0.5 µM, Biozol)
DMSO (0.05%, afatinib solvent)
Trastuzumab solvent (described in [17 (link)])
Cetuximab solvent (8.48 mg/ml NaCl, 1.88 mg/ml Na2HPO4 × 7H2O, 0.41 mg/ml NaH2PO4xH2O)
Recombinant HBEGF (5 ng/ml)
Recombinant AREG (15 ng/ml)

dependent variables

Cell proliferation

control variables

Cell numbers (see Table S1, Additional file 1)
Treatment duration (72 h)

controls

Positive control: not specified
Negative control: DMSO (0.05%, afatinib solvent)

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