Journal Article

Adenovirus infection promotes the formation of glioma stem cells from glioblastoma cells through the TLR9/NEAT1/STAT3 pathway

Liang, Liang, Zheng, Min-Hua, Zhang, Yu-Fei, Cao, Xiu-Li, Zhang, Yi-Zhe, Han, Hua, Shi, Mei, Zang, Jian, Gao, Xiang-Yu, Cao, Yuan

2020

Overview

Background Glioma stem cells (GSCs) are glioma cells with stemness and are responsible for a variety of malignant behaviors of glioma. Evidence has shown that signals from tumor microenvironment (TME) enhance stemness of glioma cells. However, identification of the signaling molecules and underlying mechanisms has not been completely elucidated. Methods Human samples and glioma cell lines were cultured in vitro to determine the effects of adenovirus (ADV) infection by sphere formation, RT-qPCR, western blotting, FACS and immunofluorescence. For in vivo analysis, mouse intracranial tumor model was applied. Bioinformatics analysis, gene knockdown by siRNA, RT-qPCR and western blotting were applied for further mechanistic studies. Results Infection of patient-derived glioma cells with ADV increases the formation of tumor spheres. ADV infection upregulated stem cell markers and in turn promoted the capacities of self-renewal and multi-lineage differentiation of the infected tumor spheres. These ADV infected tumor spheres had stronger potential to form xenograft tumors in immune-compromised mice. GSCs formation could be promoted by ADV infection via TLR9, because TLR9 was upregulated after ADV infection, and knockdown of TLR9 reduced ADV-induced GSCs. Consistently, MYD88, as well as total STAT3 and phosphorylated (p-)STAT3, were also upregulated in ADV-induced GSCs. Knockdown of MYD88 or pharmaceutical inhibition of STAT3 attenuated stemness of ADV-induced GSCs. Moreover, we found that ADV infection upregulated lncRNA NEAT1. Knockdown of NEAT1 impaired stemness of ADV-induced GSCs. Lastly, HMGB1, a damage associated molecular pattern (DAMP) that triggers TLR signaling, also upregulated stemness markers in glioma cells. Conclusion ADV, which has been developed as vectors for gene therapy and oncolytic virus, promotes the formation of GSCs via TLR9/NEAT1/STAT3 signaling. 3M_UHBD3-jpk5pbfpFGQrF Video abstract

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Adenovirus / Adenovirus diseases / Adenoviruses / Antibodies / Bioinformatics / Biomedical and Life Sciences / Biotechnology / Brain cancer / Brain tumors / Cell Biology / Cell differentiation / Cell self-renewal / Cellular signal transduction / China / Complications and side effects / Cytokines and Growth Factors / DAMP / Development and progression / Diseases / Experiments / Expression vectors / Flow cytometry / Gene therapy / Genetic aspects / Genetic vectors / Glioblastoma cells / Glioblastoma multiforme / Glioma / Glioma cells / Glioma stem cells / Gliomas / Growth factors / Health aspects / HMGB1 protein / Immunofluorescence / Infections / Life Sciences / Medical research / MyD88 protein / NEAT1 / Oncolysis / Penicillin / Protein-Ligand Interactions / Proteins / Receptors / RNA / siRNA / Spheres / Stat3 protein / Stem cell transplantation / Stem cells / TLR9 / TLR9 protein / Toll-like receptors / Tumors / Viral infections / Western blotting / Xenografts