Simultaneous detection of the concentration variations of microRNA-221 (miRNA-221) and PTEN mRNA molecules in the PI3K/AKT signaling pathway is of significance to elucidate cancer cell migration and invasion, which is useful for cancer diagnosis and therapy. In this work, we show the biodegradable MnO 2 nanosheet-assisted and target-triggered DNAzyme recycling signal amplification cascaded approach for the specific detection of the PI3K/AKT signaling pathway in live cells via simultaneous and sensitive monitoring of the variation of intracellular miRNA-221 and PTEN mRNA. Our nanoprobes enable highly sensitive and multiplexed sensing of miRNA-221 and PTEN mRNA with low detection limits of 23.6 and 0.59 pM in vitro, respectively, due to the signal amplification cascades. Importantly, the nanoprobes can be readily delivered into cancer cells and the MnO 2 nanosheets can be degraded by intracellular glutathione to release the Mn 2+ cofactors to trigger multiple DNAzyme recycling cycles to show highly enhanced fluorescence at different wavelengths to realize sensitive and multiplexed imaging of PTEN mRNA and miRNA-221 for detecting the PI3K/AKT signaling pathway. Moreover, the regulation of PTEN mRNA expression by miRNA-221 upon stimulation by various drugs can also be verified by our method, indicating its promising potentials for both disease diagnosis and drug screening.
Keyphrases
- pi k akt
- signaling pathway
- cell cycle arrest
- induced apoptosis
- label free
- cell proliferation
- epithelial mesenchymal transition
- loop mediated isothermal amplification
- binding protein
- endoplasmic reticulum stress
- living cells
- high resolution
- single cell
- drug delivery
- air pollution
- particulate matter
- quantum dots
- young adults
- emergency department
- mass spectrometry
- nucleic acid
- lymph node metastasis
- squamous cell carcinoma
- squamous cell
- sensitive detection
- gold nanoparticles
- reduced graphene oxide