RESUMO
BACKGROUND: Glioblastoma is one of the most serious brain cancer. Previous studies have demonstrated that PTEN function disorder affects the causing and exacerbation of glioblastoma. Newcastle disease virus (NDV) has been studied as a cancer virotherapeutics. In this study, PTEN gene was delivered to glioblastoma by recombinant NDV (rNDV) and translated into protein at the cytoplasm of the glioblastoma. METHODS: We did comparison tests PTEN protein expression efficiency and oncolytic effect depend on the PTEN gene insertion site at the between NP and P genes and the between P and M gene. PTEN protein mRNA transcription, translation in glioblastoma cell, and functional PTEN protein effect of the rNDV in vitro and in vivo test performed using western blotting, RT-qPCR, MTT assay, and Glioblastoma xenograft animal model test. RESULTS: The result of this study demonstrates that rNDV-PTEN kills glioblastoma cells and reduces cancer tissue better than rNDV without the PTEN gene. In molecular immunological and cytological assays, PTEN expression level was high at located in the between NP and P gene, and PTEN gene was successfully delivered to the glioblastoma cell using rNDV and PTEN gene translated to functional protein and inhibits hTERT and AKT gene. CONCLUSIONS: PTEN gene enhances the oncolytic effect of the rNDV. And our study demonstrated that NP and P gene site is better than P and M gene site which is commonly and conventionally used. PTEN gene containing rNDV is a good candidate virotherapeutics for glioblastoma.
Assuntos
Neoplasias Encefálicas , Glioblastoma , Vírus da Doença de Newcastle , Terapia Viral Oncolítica , PTEN Fosfo-Hidrolase , Animais , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/terapia , Neoplasias Encefálicas/virologia , Linhagem Celular Tumoral , Xenoenxertos , Humanos , Camundongos , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/metabolismo , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/metabolismo , Monoéster Fosfórico Hidrolases/metabolismoRESUMO
A novel immunosensor for detecting Newcastle disease virus (NDV) was developed using excessively tilted fiber grating (Ex-TFG) coated with gold nanospheres (AuNs). AuNs were coated on the Ex-TFG surface via Au-S bonds using 3-mercaptopropyltrimethoxysilane (MPTMS), and the activated staphylococcal protein A (SPA) was linked to AuNs by covalent bonds via cysteamine. AuNs greatly enhanced the impact of the analyte on the fiber cladding mode through the local surface Plasmon resonance (LSPR) effect, thus improving the detection limit and sensitivity of the immunosensor. Meanwhile, SPA enhanced the bioactivity of anti-NDV monoclonal antibodies (MAbs), thus promoting the effectiveness of specific binding events on the fiber surface. Immunoassays were performed by monitoring the resonance wavelength shift of the proposed sensor under NDV samples containing different particle amounts. Specificity was assessed, and clinical tests for NDV were performed by contrast experiments. Experimental results showed that the detection limit for NDV was about 5~10 times improved compared to that of reference Ex-TFG without AuN treatment. Moreover, the novel biosensor was reusable and could potentially be applied in clinic.