Background
We assess the effects exerted by CRISPR/Cas9 mediated microRNA 21 (miR-21) depletion on the biologic characteristics of CNE2 nasopharyngeal carcinoma (NPC) cells and the underlying mechanisms.
Conclusions
miR-21 depletion can suppress the cell growth as well as proliferation and induce apoptosis in CNE2 cells possibly by inhibiting the PI3K/AKT/mTOR signaling pathway.
Methods
The sgRNA was designed targeted at miR-21 gene, along with the construction of the CRISPR/Cas9 lentivirus system and the detection of editing efficiency through T7EN1 enzyme digestion. Effects of miR-21 depletion on the biologic characteristics of CNE2 cells were detected through CCK-8, Transwell Invasion Assay and flow cytometry. Mechanistic studies were based on bioinformatic analysis and immunoblotting.
Results
A CRISPR/Cas9 system with targeted knockdown of miR-21 gene was obtained. miR-21 depletion evidently inhibited the growth, clone formation, and invasion as well as migration abilities of CNE2 cells, thus inducing apoptosis. A total of 28 KEGG were identified through the bioinformatic analysis. Further immunoblotting showed that the expressions of proteins involved in the PI3K/AKT/mTOR signaling pathway were decreased in response to miR-21 depletion. Conclusions: miR-21 depletion can suppress the cell growth as well as proliferation and induce apoptosis in CNE2 cells possibly by inhibiting the PI3K/AKT/mTOR signaling pathway.