Abstract 3788: Profiling of microRNAs in androgen independent prostate cancer cells
Authors: Sharad Shashwat , Li Hua 
Published: April 2023

Introduction: Short noncoding microRNAs (miRNAs) function as tumor suppressors or oncogenes playing important roles in tumorigenesis and metastasis. In prostate cancer, the studies had indicated the potential roles of miRNA as biomarkers for earlier detection, diagnosis, prognosis evaluation and therapy targets. The castration resistance and metastasis remain the major challenges for the administration of prostate cancer patients, and the underlying biology is still not fully understood. Here, we reported the comparative profiling of miRNAs in hormone dependent and independent prostate cancer cells, aiming to identify novel miRNAs for monitoring aggressive disease progression and therapy targets.

Methods: The hormone sensitive LNCaP cells, and androgen independent C42B and PC3 cells were used to miRNA sequencing. The miRNA sequencing libraries were prepared with NanoDrop ND-1000. The DNA fragments were denatured to generate single-stranded DNA molecules and amplified in situ with TruSeq Rapid SR Cluster kit. Sequencing was performed using the Illumina NextSeq 500. Raw sequencing data passing the Illumina chastity filter were used for further analysis. Trimmed reads were aligned to reference genome with bowtie software. The expression levels of miRNAs were calculated using mirdeep2. Differentially expressed miRNAs analysis were performed with R package edgeR. miRNA target prediction was filtered based on miRNA Target databases. The analysis of Gene Oncology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were further conducted for miRNA biological function assessments.

Results: Among 777 known miRNAs, high expression (CPM&gt;30) of 356, 325 and 358 miRNAs were detected in LNCaP, C42B and PC3 cells, respectively. No expression (CPM=0) of 93, 114 and 75 miRNAs were detected in LNCaP, C42B and PC3 cells, respectively. Only miR-1247-3p was detected with high expression in LNCaP cells but no expression in C42B cells. Furthermore, there were 19 miRNAs with high expression in PC3 cells, and 20 miRNAs with high expression in PC3 cells but no expression in C42B cells including miR-125b-1-3p, miR-181a-3p, miR-31-3p, miR-1247-3p and miR-873-5p. In contrast, only 5 miRNAs with high expression in LNCaP cells and 7 miRNAs with high expression in C42B cells but not detectable in PC3 cells such as miR-141-5p. Pathway analysis further revealed that the miRNA regulated pathway of TGF-β, Hippo and Wnt were activated and the pathway of FoxO, P53, MAPK and Hedgehog were suppressed in C42B cells compared to LNCaP cells. Similarly, the pathway of HIF-1 was activated and the pathways of PI3K-Akt, ErbB and GnRH were inhibited in PC3 cells.

Conclusion: The sequencing of miRNA of hormone dependent and independent prostate cancer cells generated the distinct miRNA signatures which granted further validation in human prostate cancer samples and underlying biology study in aggressive disease progression.

Citation Format: Shashwat Sharad, Hua Li. Profiling of microRNAs in androgen independent prostate cancer cells. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3788.