Overexpression of lncRNA TUG1 enhances the efficacy of DC-CIK immunotherapy in neuroblastoma in vitro and in vivo

Article type: Research Article

Authors: Tan, Wei-Qiang^{a; 1} | Yuan, Li^{a; 1} | Cao, Xu^{b; 1} | Wu, Xiao-Yuan^a | Xing, Yi-Qun^a | Ye, Ming^{a; *}

Affiliations: [a] Department of Surgery, Xiang’an Hospital of Xiamen University, Xiamen University, Xiamen, Fujian, China | [b] Department of Surgery, Children’s Hospital of Soochow University, Soochow University, Suzhou, Jiangsu, China

Correspondence: [*] Corresponding author: Ming Ye, Department of Surgery, Xiang’an Hospital of Xiamen University, Xiamen University, Xiamen 361102, Fujian, China. E-mail: [email protected].

Note: [1] Wei-Qiang Tan, Li Yuan and Xu Cao are the co-first authors.

Abstract: BACKGROUND: Long non-coding RNA (LncRNA) TUG1 plays a critical role in the development of human cancers. This study explored whether TUG1 is involved in the cytotoxicity of dendritic cells and cytokine-induced killer cells (DCs-CIK), an immunotherapy approach, in neuroblastoma. METHODS: A TUG1 expression plasmid was transfected into DCs. Neuroblastoma SK-N-SH cells were incubated with CIK cells, DCs-CIK cells, and TUG1-overexpressing DCs-CIK cells, with or without irradiation. SK-N-SH cell viability, colony formation, migration, and apoptosis were analyzed using CCK-8, colony formation assay, transwell assay, and flow cytometry, respectively. Production of IL-12, IL-2 and IFN-γ in the supernatants was determined using ELISA. A dual luciferase activity assay was performed to confirm the molecular interactions between TUG1 and miR-204. Tumor-bearing mice were established by injection of SK-N-SH cells followed by stimulation with CIK cells, DC-CIK cells, and TUG1-overexpressing DCs-CIK cells. RESULTS: Compared to CIK alone or DC-CIK therapy, overexpression of TUG1 significantly suppressed tumor cell proliferation, colony formation, and migration of neuroblastoma cells. Moreover, upregulation of TUG1 robustly induced apoptosis and altered key molecules associated with apoptosis and epithelial-mesenchymal transition. Contents of IL-12, IL-2 and IFN-γ were dramatically elevated in the supernatants in the coculturing system upon transfection with TUG1. In addition, TUG1 was found to be act as miR-204 sponge. Furthermore, in vivo experiments demonstrated that upregulation of TUG1 potentiated the antitumor activity of DC-CIK immunotherapy. CONCLUSION: Overexpression of TUG1 promotes DC maturation and enhances CIK cytotoxicity, suggesting that TUG1 may be a novel target for enhancing DC-CIK based immunotherapy for neuroblastoma.

Keywords: Neuroblastoma, immunotherapy, lncRNA TUG1

DOI: 10.3233/CBM-210436

Journal: Cancer Biomarkers, vol. 36, no. 1, pp. 53-61, 2023