Abstract
Extracellular vesicles (EVs) contain various types of molecules including micro-RNAs, so isolating EVs can be an effective way to analyze and diagnose diseases. A lot of micro-RNAs have been known in relation to prostate cancer (PCa), and we evaluate miR-21, miR-141, and miR-221 in EVs and compare them with prostate-specific antigen (PSA). EVs were isolated from plasma of 38 patients with prostate cancer and 8 patients with benign prostatic hyperplasia (BPH), using a method that showed the highest recovery of RNA. Isolation of EVs concentrated micro-RNAs, reducing the cycle threshold (Ct) value of RT-qPCR amplification of micro-RNA such as miR-16 by 5.12 and miR-191 by 4.65, compared to the values before EV isolation. Normalization of target micro-RNAs was done using miR-191. For miR-221, the mean expression level of patients with localized PCa was significantly higher than that of the control group, having 33.45 times higher expression than the control group (p < 0.01). Area under curve (AUC) between BPH and PCa for miR-221 was 0.98 (p < 0.0001), which was better than AUC for prostate-specific antigen (PSA) level in serum for the same patients. The levels of miR-21 and miR-141 in EVs did not show significant changes in patients with PCa compared to the control group in this study. This study suggests isolating EVs can be a helpful approach in analyzing micro-RNAs with regard to disease.
Highlights
Micro-RNAs are small noncoding RNA molecules that regulate gene expression at the posttranscriptional level [1]
The aim of this study is to investigate how micro-RNAs in extracellular vesicles (EVs) would be effective in comparison with prostate-specific antigen (PSA), and we test three miRNAs, miR-21, miR-141, and miR-221 by isolating EVs from plasma, and compare the result with the level of PSA regarding their diagnostic performance in patients with prostate cancer [18, 19]
In the receiver operating characteristic curve (ROC) about the predictability for prostate cancer, the area under the curve (AUC) for miR-221 in EVs from plasma was 0.98, while Area under curve (AUC) for PSA was 0.86 (Fig 5B)
Summary
Micro-RNAs (miRNAs) are small noncoding RNA molecules that regulate gene expression at the posttranscriptional level [1]. They play a regulatory role by binding to 3’ UTR of transcripts and are involved in many developmental and physiological processes [2]. Abnormal expression pattern of micro-RNAs has been related to disease such as cancer, and lots of research has been conducted to apply micro-RNAs diagnostically [3] In biological fluids such as blood, urine, and saliva [4,5,6], micro-RNAs are known to exist in one of the forms that involve extracellular vesicles (EVs) or proteins like high-density lipoprotein or Argonaute protein [7, 8].
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