ABSTRACT
Recently, PIWI-interacting small non-coding RNAs (piRNAs) have emerged as novel cancer biomarkers candidate because of their high expression level in various cancer types and role in the control of tumor suppressor genes. In this study, a novel breast cancer theragnostics probe based on a single system targeting the piRNA-36026 (piR-36026) molecular pathway was developed using a piR-36026 molecular beacon (MB). The piR-36026 MB successfully visualized endogenous piR-36026 biogenesis, which is highly expressed in MCF7 cells (a human breast cancer cell line), and simultaneously inhibited piR-36026-mediated cancer progression in vitro and in vivo. We discovered two tumor suppressor proteins, SERPINA1 and LRAT, that were directly regulated as endogenous piR-36026 target genes in MCF7 cells. Furthermore, multiplex bioimaging of a single MCF7 cell following treatment with piR-36026 MB clearly visualized the direct molecular interaction of piRNA-36026 with SERPINA1 or LRAT and subsequent molecular therapeutic responses including caspase-3 and PI in the nucleus.
Subject(s)
Breast Neoplasms/genetics , Gene Expression Regulation, Neoplastic , RNA, Small Interfering/genetics , Acyltransferases/genetics , Animals , Base Sequence , Breast/diagnostic imaging , Breast/metabolism , Breast/pathology , Breast Neoplasms/diagnostic imaging , Breast Neoplasms/pathology , Female , HEK293 Cells , Humans , MCF-7 Cells , Male , Mice, Inbred BALB C , Mice, Nude , Oligonucleotide Probes/analysis , Oligonucleotide Probes/genetics , Optical Imaging , RNA, Small Interfering/analysis , alpha 1-Antitrypsin/geneticsABSTRACT
Current methods for delivering foreign genetic materials into mammalian cells are highly successful. However, these methods cannot be applied in oocyte or embryo systems due to their toxicity and low efficiency. Moreover, no satisfactory methods exist for delivering foreign genetic material without inducing physical damage to membranes. Here we developed an organic compound (VisuFect)-mediated small interfering RNA (siRNA) delivery method and evaluated this method in P19 cells and mouse zygotes. Oct4-siRNA conjugated VisuFect (Oct4-siRNA-VF) permeated the zona pellucida effectively and localized inside mouse zygotes without inducing membrane damage. Successful VisuFect-mediated delivery was further demonstrated by strong transcriptional repression of Oct4 expression by the delivered Oct4-siRNA, in addition to repressed embryonic development of mouse zygotes.
Subject(s)
Gene Transfer Techniques , RNA, Small Interfering/administration & dosage , Zygote , Animals , Embryonic Development , Female , Mice , Octamer Transcription Factor-3/biosynthesis , Octamer Transcription Factor-3/genetics , Pregnancy , Zona PellucidaABSTRACT
A self-assembling magnetic resonance beacon was used to visualize the microRNA-1 expression-dependent change in magnetic resonance signal intensity.
Subject(s)
MicroRNAs/analysis , MicroRNAs/chemistry , Oligonucleotide Probes/chemistry , Animals , Cell Line , Fluorescence Resonance Energy Transfer , Magnetic Resonance Spectroscopy , Mice , Nanoparticles/chemistryABSTRACT
A dual fluorophore-based color-tunable molecular beacon visualized the microRNA-294 expression-dependent color change in cells.
