Introduction: Prostate cancer ranks second globally in cancer diagnoses and fifth in cancer deaths among men.1 This study aims to correlate microRNA (miR) concentration profiles in prostate cancer cell lines with stage. miR, 20-22 long non-coding RNA, holds promise for accurate prostate cancer prognosis compared to current methods like prostate-specific antigen (PSA) and prostate biopsy. 2 miR concentrations could more accurately correlate circulating biomarkers in prostate cancer cells with progression.2 RT-qPCR is the most sensitive technique for miR detection and quantification.2 This project aims to validate multiple miR concentrations in prostate cancer cells and correlate with disease progression by cell type. We found cell lines DU145, PC3, and 22RV1 have higher levels of miR-1290 concentrations. Statistical analysis through t-test show that castration-resistant prostate cancer (CRPC) cell line 22RV1 has higher miR-1290 concentrations.
Materials and
Methods: Cell lines DU145, PC3, and 22RV1 were selected based on stage and severity of prostate cancer with HEK 293 cells as a negative control. Standard cell culture protocol was performed. Next, RNA extraction and isolation occurred followed by two step RT-qPCR. Nanodrop results of samples were taken to test purity of total RNA collected to ensure RT-qPCR was performed properly. The student’s t-test will look at the significance of difference concentrations across cell lines. The initial objective was to develop miR standard assays for target miR-375 and miR-1290, alongside internal controls (miR-16 and miR-30a-5p) by adapting existing methods (standard assays). Our second objective was to quantify the miR concentrations in five prostate cancer cell lines using RT-qPCR and assess correlation across different PCa stages.
Results, Conclusions, and Discussions: We found 22RV1 cells have higher miR-1290 concentrations. Total RNA concentration raw data is shown in Fig 1. A larger N value for cell lines to see the clear difference between PC3 and DU145 and rank cell lines by severity level (22RV1 > PC3 > DU145 > LNCAP in terms of stages). Data shows that 22RV1 and DU145 will have higher target concentration based on Fig 2. Statistical analysis shows that cell line 22RV1 has higher miR-1290 concentrations. This project aims to develop RT-PCR assays to accurately and rapidly quantify miR expression profiles from prostate cancer cell lines. In the future, we have identified another negative control with HeLa cells and will continue to test if controls are stably expressed across cell lines (p-test, p-value ≤ 0.05).
Acknowledgements (Optional): We thank Smith lab, the Illinois Scholars Undergraduate Research (ISUR) Program, and the Cancer Center of Illinois (CCIL). We thank the University of Illinois Urbana-Champaign Department of Bioengineering, for fostering an outstanding academic environment.
