LncRNA HAS2-AS1 Promotes Glioblastoma Proliferation by Sponging miR-137

Yalin Lu; Gaochao Guo; Rujun Hong; Xingjie Chen; Yan Sun; Fang Liu; Zhimeng Zhang; Xun Jin; Jun Dong; Kai Yu; Xuejun Yang; Yang Nan; Qiang Huang

doi:10.3389/fonc.2021.634893

LncRNA HAS2-AS1 Promotes Glioblastoma Proliferation by Sponging miR-137

Front Oncol. 2021 May 20:11:634893. doi: 10.3389/fonc.2021.634893. eCollection 2021.

Authors

Yalin Lu^{1

2

3}, Gaochao Guo⁴, Rujun Hong^{1

2

3}, Xingjie Chen^{1

2

3}, Yan Sun^{1

2

3}, Fang Liu⁵, Zhimeng Zhang⁶, Xun Jin^{7

8

9

10}, Jun Dong¹¹, Kai Yu^{1

2

3}, Xuejun Yang^{1

2

3}, Yang Nan^{1

2

3

12}, Qiang Huang^{1

2

3

12}

Affiliations

¹ Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, China.
² Key Laboratory of Post-Trauma Neuro-Repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, China.
³ Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China.
⁴ Department of Neurosurgery, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, Zhengzhou, China.
⁵ Department of Psychiatry and Imaging-Genetics and Co-morbidity (PNGC Lab), Tianjin Anding Hospital, Tianjin Mental Health Center, Mental Health Teaching Hospital, Tianjin Medical University, Tianjin, China.
⁶ Department of Neurosurgery, Ningbo Hospital of Zhejiang University, Ningbo, China.
⁷ Tianjin Medical University Cancer Institute and Hospital, Tianjin, China.
⁸ National Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, Tianjin, China.
⁹ Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University Cancer Institute and Hospital, Tianjin, China.
¹⁰ Tianjin's Clinical Research Center for Cancer, Tianjin, China.
¹¹ Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Soochow, China.
¹² Department of Neurosurgery, Tianjin Medical University General Hospital Airport Site, Tianjin, China.

Abstract

GBM (Glioblastoma multiform) is the most malignant tumor type of the central nervous system and has poor diagnostic and clinical outcomes. LncRNAs (Long non-coding RNAs) have been reported to participate in multiple biological and pathological processes, but their underlying mechanism remains poorly understood. Here, we aimed to explore the role of the lncRNA HAS2-AS1 (HAS2 antisense RNA 1) in GBM. GSE103227 was analyzed, and qRT-PCR was performed to measure the expression of HAS2-AS1 in GBM. FISH (Fluorescence in situ hybridization) was performed to verify the localization of HAS2-AS1. The interaction between HAS2-AS1 and miR-137 (microRNA-137) was predicted by LncBook and miRcode followed by dual-luciferase reporter assays, and the relationships among HAS2-AS1, miR-137 and LSD1 (lysine-specific demethylase 1) were assessed by WB (western blot) and qRT-PCR. Colony formation and CCK-8 (cell counting kit-8) assays were performed as functional tests. In vivo, nude mice were used to confirm the function of HAS2-AS1. HAS2-AS1 expression was upregulated in GBM cell lines, and HAS2-AS1 was localized mainly in the cytoplasm. In vitro, high HAS2-AS1 expression promoted proliferation, and knockdown of HAS2-AS1 significantly inhibited proliferation. Furthermore, HAS2-AS1 functioned as a ceRNA (competing endogenous RNA) of miR-137, leading to the disinhibition of its downstream target LSD1. The miR-137 level was downregulated by HAS2-AS1 overexpression and upregulated by HAS2-AS1 knockdown. In a subsequent study, LSD1 expression was negatively regulated by miR-137, while miR-137 reversed the LSD1 expression levels caused by HAS2-AS1. These results were further supported by the nude mouse tumorigenesis experiment; compared with xenografts with high HAS2-AS1 expression, the group with low levels of HAS2-AS1 exhibited suppressed proliferation and better survival. We conclude that lncRNA HAS2-AS1 promotes proliferation by functioning as a miR-137 decoy to increase LSD1 levels and thus might be a possible biomarker for GBM.

Keywords: LSD1; ceRNA; glioblastoma; lncRNA; miR-137.