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Bioengineered miR-34a modulates mitochondrial inner membrane protein 17 like 2 (MPV17L2) expression toward the control of cancer cell mitochondrial functions
- Source :
- Bioengineered, vol 13, iss 5
- Publication Year :
- 2022
- Publisher :
- Taylor & Francis, 2022.
-
Abstract
- Genome-derived microRNAs (miRNAs or miRs) control post-transcriptional gene expression critical for various cellular processes. Recently, we have invented a novel platform technology to achieve high-yield production of fully humanized, bioengineered miRNA agents (hBERAs) for research and development. This study is aimed to produce and utilize a new biologic miR-34a-5p (or miR-34a) molecule, namely, hBERA/miR-34a, to delineate the role of miR-34a-5p in the regulation of mitochondrial functions in human carcinoma cells. Bioengineered hBERA/miR-34a was produced through in vivo fermentation production and purified by anion exchange fast protein liquid chromatography. hEBRA/miR-34a was processed to target miR-34a-5p in human osteosarcoma and lung cancer cells, as determined by selective stem-loop reverse transcription quantitative polymerase chain reaction analysis. The mitochondrial inner membrane protein MPV17 like 2 (MPV17L2) was validated as a direct target for miR-34a-5p by dual luciferase reporter assay. Western blot analysis revealed that bioengineered miR-34a-5p effectively reduced MPV17L2 protein outcomes, leading to much lower levels of respiratory chain Complex I activities and intracellular ATP that were determined with specific assay kits. Moreover, Seahorse Mito Stress Test assay was conducted, and the results showed that biologic miR-34a-5p sharply reduced cancer cell mitochondrial respiration capacity, accompanied by a remarkable increase of oxidative stress and elevated apoptotic cell death, which are manifested by greater levels of reactive oxygen species and selective apoptosis biomarkers, respectively. These results demonstrate the presence and involvement of the miR-34a-5p-MPV17L2 pathway in the control of mitochondrial functions in human carcinoma cells and support the utility of novel bioengineered miRNA molecules for functional studies.
- Subjects :
- Lung Neoplasms
1.1 Normal biological development and functioning
cancer metabolism
Bone Neoplasms
Bioengineering
Microbiology
Applied Microbiology and Biotechnology
Mitochondrial Proteins
Rare Diseases
Underpinning research
Genetics
Humans
MPV17L2
Cell Proliferation
miRNA
Cancer
Biological Products
Bioengineered
Carcinoma
Membrane Proteins
General Medicine
Mitochondria
MicroRNAs
Generic health relevance
miR-34a
Biotechnology
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Bioengineered, vol 13, iss 5
- Accession number :
- edsair.doi.dedup.....66dd043e42ae49d3ffe86958d67a921a
- Full Text :
- https://doi.org/10.6084/m9.figshare.19779799.v1