The phosphatase and tensin homolog gene inserted between NP and P gene of recombinant Newcastle disease virus oncolytic effect test to glioblastoma cell and xenograft mouse model

Abstract

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.

The glioblastoma suppression effect of recombinant Newcastle disease virus harboring the PTEN gene delivered intravenously to a U87 MG PTEN mutant glioblastoma cell-bearing orthotropic mouse model

“…Many studies have reported that overexpression of PTEN in GBM reduces cell proliferation and increases cell death by inactivating the Akt pathway (31464538, 32445778). We prepared recombinant rNDV using a NDV backbone for PTEN overexpression and used it to infect U87 MG GBM cells; rNDV replication e ciency and PTEM overexpression were con rmed in these cells 9 . Overexpression of PTEN signi cantly decreased the proliferation of U87 MG cells compared with SH-SY5Y neuroblastoma cells (Fig.…”

“…The recombinant NDV containing the PTEN gene (rNDV-PTEN) virus was previously constructed 9 . The viruses were propagated by infection of vero cells at an MOI of 0.5 for 2 days.…”

“…The following antibodies: anti-GAPDH (sc-32233) (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-LC3 (NB100-2220, Novos Bio, USA); anti-MMP9 (MA5-15886, ThermoFisher); anti-PCNA (PC 10, Sigma-Aldrich, USA); anti-P-mTOR (Ser2448, 2971S), anti-mTOR (2972S), anti-P-Akt (Ser473, 9271S), anti-Akt (9272S), anti-cleaved Cas9 (9509S), anti-cleaved Cas3 (9664S), anti-cleaved Cas8 (9496S), anti-Bax (2772S), anti-p62 (5114S) and anti-PTEN (9552S) (Cell signaling Technology, Danvers, MA, USA). Immunoblotting of 30 µg of whole-cell lysate or tissue homogenate and next step were performed as described previously 9 . Values were normalized to GAPDH as loading controls.…”

“…We previously constructed recombinant NDV expressing human PTEN (rNDV-PTEN) and showed that rNDV-PTEN inhibits the growth of GBM cells in vitro and in a xenograft animal model 9 . In this study, we evaluated whether rNDV-PTEN can increase PTEN expression and induce an antitumor effect in an orthotopic GBM mouse model.…”

The tumor suppressive effect and apoptotic mechanism of TRAIL gene‐containing recombinant NDV in TRAIL‐resistant colorectal cancer HT‐29 cells and TRAIL‐nonresistant HCT116 cells, with each cell bearing a mouse model

“…Lytic strains of NDV can infect and kill cancer cells as well as release newly formed progeny viruses 20,21 . The oncolytic effects and mechanisms of several low‐virulence NDV strains have been studied 22–24 . The cancer cell‐killing mechanisms of non‐lytic NDV involve both intrinsic and extrinsic apoptotic pathways and stimulation of immune responses against NDV‐infected cancer cells.…”