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Updated: Jan 3, 2026
Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
Multiplex profiling identifies clinically relevant signalling proteins in an isogenic prostate cancer model of
S Inder1,2, M Bates1, N Ni Labhrai1
1Translational Radiobiology and Molecular oncology, Applied Radiation Therapy Trinity, Trinity Translational Medicine Institute (TTMI), Trinity College Dublin, Dublin, Ireland.
Abstract:
The exact biological mechanism governing the radioresistant phenotype of prostate tumours at a high risk of recurrence despite the delivery of advanced radiotherapy protocols remains unclear. This study analysed the protein expression profiles of a previously generated isogenic 22Rv1 prostate cancer model of radioresistance using DigiWest multiplex protein profiling for a selection of 90 signalling proteins. Comparative analysis of the profiles identified a substantial change in the expression of 43 proteins. Differential PARP-1, AR, p53, Notch-3 and YB-1 protein levels were independently validated using Western Blotting. Pharmacological targeting of these proteins was associated with a mild but significant radiosensitisation effect at 4Gy. This study supports the clinical relevance of isogenic in vitro models of radioresistance and clarifies the molecular radiation response of prostate cancer cells.
Insights
Prostate tumors resisting radiation therapy show altered protein levels, including PARP-1 and AR. Targeting these proteins may improve radiotherapy effectiveness for recurrent prostate cancer.
Area of Science:
- Oncology
- Molecular Biology
- Radiotherapy Research
Background:
- Prostate tumors can resist radiotherapy, leading to recurrence despite advanced treatments.
- The underlying biological mechanisms of this radioresistant phenotype remain largely unknown.
Purpose of the Study:
- To investigate the protein expression profiles in an isogenic model of prostate cancer radioresistance.
- To identify key signaling proteins involved in the molecular response to radiation.
Main Methods:
- Utilized DigiWest multiplex protein profiling on 90 signaling proteins in a 22Rv1 prostate cancer radioresistance model.
- Validated differential protein expression of key candidates (PARP-1, AR, p53, Notch-3, YB-1) using Western Blotting.
Main Results:
- Identified substantial changes in 43 proteins, with significant differences in PARP-1, AR, p53, Notch-3, and YB-1 expression.
- Pharmacological targeting of these identified proteins resulted in mild but significant radiosensitization at 4Gy.
Conclusions:
- This study validates the clinical relevance of isogenic in vitro models for studying radioresistance.
- The findings clarify the molecular mechanisms of radiation response in prostate cancer cells, highlighting potential therapeutic targets.
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