Open Journals System
Issue Information: Vol. 4, No. 2,(2012)

Article Title: Assessment of tissue distribution and subcellular localization of miR-302 and miR-21 by means of in situ hybridization technique

pages: 50-55

DOI: 10.22067/jcmr.v4i2.22595

MicroRNAs (miRNAs) are a group of short non-coding RNAs implicated in numerous fundamental cellular
processes, and their disregulations have been linked to several pathologic conditions, mainly cancers.
Determining tissue distribution of miRNAs is a prerequisite for understanding their exact functions during
development, tissue homeostasis and abnormality. In situ hybridization is a powerful technique to delineate the
sub-cellular localization and tissue distribution patterns of mRNAs as well as miRNAs. Due to the important
role of miRNAs in tumorigenesis, we optimized an ISH technique for detection of two well-known miRNAs
(miR-302 and miR-21) in formalin-fixed paraffin-embedded (FFPE) tumor samples along with a pluripotent
embryonal carcinoma cell line, NTERA-2 (NT2). After fixation of cells on slides/sectioning of FFPE blocks,
proteinase K digestion, probe concentration, antibody development and light sensitive color reaction were
optimized for both the FFPE samples and cell line. Signals for U6 snRNA, as an internal control, were detected
in the nuclei of the cells. MiR-21 and miR-302 expression was detected in the cytoplasm of FFPE samples of
seminoma carcinoma and in NT2 cell line, respectively. In this study, we optimized ISH for miRNA detection
in FFPE samples and NT2 cell line.

key words:   in situ hybridization; microRNA; miR-302; miR-21

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