Abstract
BackgroundThe microRNA‐371a‐3p (miR‐371a‐3p) has been reported to be an informative liquid biopsy (serum and plasma) molecular biomarker for both diagnosis and follow‐up of patients with a malignant (testicular) germ cell tumor ((T)GCT). It is expressed in all histological cancer elements, with the exception of mature teratoma. However, normal testis, semen, and serum of males with a disrupted testicular integrity without a TGCT may contain miR‐371a‐3p levels above threshold, of which the cellular origin is unknown.ObjectivesTherefore, a series of relevant tissues (frozen and formalin‐fixed paraffin‐embedded (FFPE), when available) from the complete male urogenital tract (i.e., kidney to urethra and testis to urethra) and semen was investigated for miR‐371a‐3p levels using targeted quantitative RT‐PCR (qRT‐PCR).Materials and methodsIn total, semen of males with normospermia (n = 11) and oligospermia (n = 3) was investigated, as well as 88 samples derived from 32 different patients. The samples represented one set of tissues related to the entire male urogenital tract (11 anatomical locations), three sets for 10 locations, and four sets for six locations.ResultsAll testis parenchyma (n = 17) cases showed low miR‐371a‐3p levels. Eight out of 14 (57%) semen samples showed detectable miR‐371a‐3p levels, irrespective of the amount of motile spermatozoa, but related to sperm concentration and matched Johnsen score (Spearman's rho correlation coefficient 0.849 and 0.871, p = 0.000, respectively). In all other tissues investigated, miR‐371a‐3p could not be detected.DiscussionThis study demonstrates that the miR‐371a‐3p in healthy adult males is solely derived from the germ cell compartment.ConclusionsThe observation is important in the context of applying miR‐371a‐3p as molecular liquid biopsy biomarker for diagnosis and follow‐up of patients with malignant (T)GCT. Moreover, miR‐371a‐3p might be an informative seminal biomarker for testicular germ cell composition.
Highlights
IntroductionMicroRNAs (miRNAs) are small, non-coding single-strandedRNA molecules about 22 nucleotides long that are involved in post-transcriptional gene regulation (Lee et al, 1993; Reinhart et al, 2000; Bentwich et al, 2005; Zamore & Haley, 2005).miRNAs are found in diverse organisms, including animals and plants (Ambros, 2003), and are highly stable in various types of human body fluid, including serum, plasma, cerebrospinal fluid, saliva, ejaculate, seminal plasma, and urine (Calin et al, 2002; Reis et al, 2010).In 2006, the relevance of a defined set of embryonic stem cellassociated miRNAs, including miR-371a-3p, was identified as potential oncogene for malignant testicular germ cell tumors (TGCT) (Voorhoeve et al, 2006)
This study indicates that in healthy males, the germ cell compartment is the cellular origin of miR-371a-3p
The cluster miR-371-373 is expressed in all histological elements of primary as well as metastatic testicular germ cell tumors (TGCT), except for pure teratoma (Voorhoeve et al, 2006; Cheng et al, 2018; Leao et al, 2018; Terbuch et al, 2018)
Summary
MicroRNAs (miRNAs) are small, non-coding single-strandedRNA molecules about 22 nucleotides long that are involved in post-transcriptional gene regulation (Lee et al, 1993; Reinhart et al, 2000; Bentwich et al, 2005; Zamore & Haley, 2005).miRNAs are found in diverse organisms, including animals and plants (Ambros, 2003), and are highly stable in various types of human body fluid, including serum, plasma, cerebrospinal fluid, saliva, ejaculate, seminal plasma, and urine (Calin et al, 2002; Reis et al, 2010).In 2006, the relevance of a defined set of embryonic stem cellassociated miRNAs, including miR-371a-3p, was identified as potential oncogene for malignant testicular germ cell tumors (TGCT) (Voorhoeve et al, 2006). The microRNA-371a-3p (miR-371a-3p) has been reported to be an informative liquid biopsy (serum and plasma) molecular biomarker for both diagnosis and follow-up of patients with a malignant (testicular) germ cell tumor ((T)GCT). It is expressed in all histological cancer elements, with the exception of mature teratoma. MiR-371a-3p might be an informative seminal biomarker for testicular germ cell composition
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