University of Twente Student Theses


Characterizing prostate cancer cell lines of varied aggressiveness and evaluating their response to Enzalutamide

Peters, Y. (2022) Characterizing prostate cancer cell lines of varied aggressiveness and evaluating their response to Enzalutamide.

[img] PDF
Abstract:Prostate cancer (PCa) is the most diagnosed cancer in men and it is the second leading cause of cancer-related death in Western societies. Despite improved treatments for metastatic prostate cancer patients, not all patients respond similarly thereby affecting overall survival. To reduce the frequent administration and ineffectiveness of treatments, there is a need for prognostic markers which can help predict the response of patients to therapies, making treatments more targeted and personalized. In this study, we aimed to develop a realistic model to study the phenotypic response of PCa cell lines, towards androgen stimulation and inhibition. We performed a proteomic analysis (PSA secretion) of multiple PCa cell lines (LNCaP, PC-3, 22Rv1, RWPE-1, RWPE-2 and PWR-1E, EMCPCa-41 organoid cells) derived from different stages of PCa, in response to androgen stimulation (R1881) and inhibition (Enzalutamide). We detected PSA secretion from different malignant PCa cell lines (LNCaP, 22Rv1, EMC-PCa-41) and observed absence of PSA secretion in the benign cell lines. This is confirmed with ELISA. In addition, PSA secretion from single LNCaP cells was identified and we observed that the PSA secretion of LNCaP cells is affected by the anti-androgen Enzalutamide. These results suggest that LNCaP cells can be used as a model mimicking the responses to anti-androgens in mHSPC patients. However, this method fails to distinguish between PSA secretion from low-producing cell lines and artifacts. To overcome this, a microwell chip is recommended and more optimization on the quantification method must be done. The PCa cell lines (LNCaP, PC-3, 22Rv1, RWPE-1, RWPE-2 and PWR-1E) were also phenotypically characterized using immunofluorescence staining (PSA, PSMA) and flow cytometry (PSMA, EpCAM, HER2). Our findings indicate PSMA, EpCAM, and HER2 expression in malignant PCa cell lines and the absence of PSMA expression in the benign cell lines. In addition, the hormone-sensitive LNCaP cell line shows higher levels of PSMA and HER2 expression compared to the benign and the castration-resistant cell lines. Proteomic analysis of single cells can help identify prognostic biomarkers and develop an effective and personalised therapy for PCa patients. Using a PVDF membrane, LNCaP cells can serve as a good model to study the PSA secretion in response to drugs. In the future, a microwell chip can help to identify the effect of drugs and stimulation on PSA secretion from the other PCa cell lines.
Item Type:Essay (Master)
Faculty:TNW: Science and Technology
Programme:Biomedical Engineering MSc (66226)
Link to this item:
Export this item as:BibTeX
HTML Citation
Reference Manager


Repository Staff Only: item control page