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MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...

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Detection of miRNA Targets in High-throughput Using the 3'LIFE Assay

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Published on: May 25, 2015

Novel microRNAs modulating ecto-5'-nucleotidase expression.

Theresa Kordaß^1,2, Tsu-Yang Chao¹, Wolfram Osen¹

¹GMP & T Cell Therapy Unit, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Frontiers in Immunology

|July 6, 2023

Summary

This study identifies microRNAs (miRNAs) that regulate immune checkpoint molecules (ICMs) like NT5E in cancer cells. Some miRNAs suppress ICMs, potentially acting as tumor suppressors, while others promote ICMs, acting as oncogenes.

Keywords:

CD274 ENTPD1 NT5E/CD73 breast cancer melanoma miRNAs

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Last Updated: Jul 24, 2025

Detection of miRNA Targets in High-throughput Using the 3'LIFE Assay

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Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library

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Published on: April 6, 2012

Area of Science:

Oncology
Molecular Biology
Immunology

Background:

Immune checkpoint molecules (ICMs) on cancer cells promote immune escape by suppressing anti-tumor immune responses.
Ecto-5'-nucleotidase (NT5E/CD73) upregulates immunosuppressive adenosine, hindering T cell-mediated tumor attack.
MicroRNAs (miRNAs) are key regulators of gene expression, and their aberrant profiles in cancer offer diagnostic and therapeutic potential.

Purpose of the Study:

To identify specific miRNAs that modulate the expression of key immune checkpoint molecules (ICMs) in human cancer cell lines.
To characterize miRNAs that either suppress or enhance ICM expression, defining potential tumor suppressors and oncogenes.
To validate the findings of miRNA-mediated ICM regulation across multiple tumor types.

Main Methods:

High-throughput screening of a human miRNA library against melanoma (SK-Mel-28) and breast cancer (MDA-MB-231) cell lines.
Assessing the impact of identified miRNAs on the expression of NT5E, ENTPD1, and CD274.
In vitro validation of miRNA effects on NT5E expression across 12 diverse cancer cell lines.

Main Results:

Identified miR-1285-5p, miR-155-5p, and miR-3134 as potent inhibitors of NT5E expression.
Discovered miR-134-3p, miR-6859-3p, miR-6514-3p, and miR-224-3p as strong enhancers of NT5E expression.
Confirmed the regulatory roles of these miRNAs in NT5E expression across various cancer cell lines.

Conclusions:

The identified miRNAs hold potential as therapeutic agents targeting ICMs in cancer.
These miRNAs may serve as valuable biomarkers for early cancer detection or prognosis.
Specific miRNAs can be therapeutically targeted to modulate ICMs and enhance anti-tumor immunity.