Nanoparticles (NPs)-meditated si-lncRNA NONHSAT159592.1 inhibits glioblastoma progression and invasion through targeting the ITGA3/FAK/PI3K/AKT pathway

Abstract

The study aims to investigate the regulatory role of NPs lncRNA NONHSAT159592.1 in glioblastoma cells and its molecular mechanism. We have designed a reduction-responsive nanoparticle (NP) platform for efficient delivery of si-lncRNA (si-lnc). The size of siRNA nanoparticles was observed and determined by transmission electron microscopy. The distribution size of nanoparticles was analyzed by the NanoSight nanoparticle tracking analyzer. The fluorescence spectrum and UV spectrum were determined. The level of lncRNA in glioblastoma cells was detected by RT-qPCR analysis. The localization of lncRNA NONHSAT159592.1 in glioblastoma cells was detected by fluorescence in situ hybridization. Cell proliferation activity was evaluated by clonal formation experiment and CCK-8 kit. Cell migration and invasion were detected by wound healing assay and Transwell experiment. Western blot assay was used to detect the expression level of EMT-related proteins in cells. EdU staining was used to detect cell proliferation. NPs or PBS and IR780 were injected intravenously into nude mice with tumors, and fluorescence imaging was performed in vivo to evaluate the proliferation of tumor tissue. The positive rate of Ki67 and Vimentin in tumor tissue was detected by immunohistochemical staining. We found that lncRNA NONHSAT159592.1 was significantly down-regulated in glioblastoma cell lines, localized in the nucleus and cytoplasm. In U87 and U251 cells, we found that NPs-si-lncRNA NONHSAT159592.1 significantly inhibited glioblastoma cell proliferation, invasion, and EMT progression. In the orthotopic xenograft model, we found that silencing lncRNA could significantly inhibit tumor proliferation and prolong the survival time of tumor-bearing mice. Further studies confirmed that overexpression of ITGA3 reversed the inhibitory effects of NPs-si-lnc on the proliferation, invasion, and migration of glioblastoma cell lines. Our study suggested that NPs (si-lnc) could inhibit the malignant development of glioma by a mechanism that may be linked to the activation of the ITGA3/FAK/PI3K/AKT signaling pathway.

Keywords: Glioblastoma; ITGA3; Nanoparticles; lncRNA NONHSAT159592.1.