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. 2021 Jul 25;12(19):5732-5744.
doi: 10.7150/jca.60257. eCollection 2021.

LncRNA-PVT1 indicates a poor prognosis and promotes angiogenesis via activating the HNF1B/EMT axis in glioma

Yongyan Bi  1   2 , Jie Ji  3 , Youxin Zhou  1
Affiliations

LncRNA-PVT1 indicates a poor prognosis and promotes angiogenesis via activating the HNF1B/EMT axis in glioma

Yongyan Bi et al. J Cancer. .

Abstract

Recent studies identified that long non-coding RNAs (lncRNAs) exhibited critical roles in tumor migration and invasion. However, the roles of lncRNAs in glioma remain unclear. The aim of this study was to uncover the underlying mechanisms of glioma progression and provide potential therapeutic targets for its treatment in clinic. Our microarray study showed that lncRNA-PVT1 was significantly upregulated in glioma tissues and played an important role in cell proliferation, migration, invasion and angiogenesis. Our data showed that the expression of lncRNA-PVT1 was increased obviously and associated with advanced tumor stage, metastasis, invasion ability, and poor prognosis in glioma patients. Up-regulation of lncRNA-PVT1 was observed to promote glioma cells proliferation, and invasion abilities in vitro as well as tumor growth in vivo by regulating miR-1207-3p expression. Online software (TargetScan, miRDB and miR TarBase) were used to predict the regulating mechanisms of lncRNA-PVT1, miR-1207-3p and HNF1B, which were validated by dual-luciferase reporter gene system. In vivo tumor-bearing mice models were established to validate the cellular results. Therefore, we suggested that lncRNA-PVT1/miR-1207-3p/HNF1B axis might play critical roles in glioma progression, indicating that lncRNA-PVT1/miR-1207-3p/HNF1B signaling axis may serve as novel molecular targets for glioma prevention and treatment.

Keywords: EMT; HNF1B; glioma; lncRNA-PVT1; miR-1207-3p.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

Figures

Figure 1
Figure 1
Dysregulated expression profiles of lncRNAs in glioma. (A) Aberrantly expression of lncRNAs was evaluated in 3 pairs of glioma and adjacent normal tissues though lncRNA expression microarray. (B) lncRNA-PVT1 expression in microarray datasets GSE29384, GSE7181 and GSE23806. (C) Scatter plot of gene expression in glioma compared with the control group (x-axis). Up-regulated lncRNAs are shown in red while down-regulated lncRNAs are shown in green. (D, E) GO biological process classification for differently expressed lncRNAs. The upper graph was related to the up-regulated lncRNAs (D) while the lower graph was related to the down-regulated lncRNAs (E). (F, G) GSEA revealed that proliferation and metastasis related biological functions were enriched in response to high lncRNA-PVT1 expression based on the microarray data set.
Figure 2
Figure 2
lncRNA-PVT1 was upregulated in glioma. (A, B) lncRNA-PVT1 expression in multiple solid tumors and normal tissues from TCGA Pan-Cancer dataset. (C) lncRNA-PVT1 expression in different TNM stage of glioma cases from TCGA dataset. (D) The lncRNA-PVT1 expression in glioma cases were determined by qRT-PCR assay. (E, F) High lncRNA-PVT1 expression was positively associated with advanced TNM stage and metastasis in patients with glioma. (G, H) The Kaplan-Meier curves introduced for survival analysis in glioma patients with high level and low level of lncRNA-PVT1. **p<0.01, ***p<0.001.
Figure 3
Figure 3
lncRNA-PVT1 promotes glioma the development of pro-metastatic phenotype in vitro. (A) lncRNA-PVT1 expression in glioma cell lines (H4, SHG-44, U87 and U251) and HEB was detected by qRT-PCR assay. (B, C) Knockdown of lncRNA-PVT1 by shRNA in U87 and U251 cells. (D-F) MTT assay was performed to assess the function of lncRNA-PVT1 in glioma cells proliferation. (G-I) The supernatants from sh-lncRNA-PVT1 cells exhibited a strong negative effect on tubule formation of HUVECs, compared to control supernatants in U87 and U251 cells. (J-L) Knockdown of lncRNA-PVT1 in U251 and U87 cells significantly inhibited cell invasion. We selected the 1# and 2# sh-plasmids with high interference efficiency for testing. *p<0.05.
Figure 4
Figure 4
lncRNA-PVT1 acted as a sponge for miR-1207-3p. (A) lncRNA-PVT1 was located in the cytoplasm of U87 and U251 cells. (B) Venn diagrams showed the results of potential targets of lncRNA-PVT1 (LncBase Predicted, Starbase 3.0 and miRcode databases). (C) Bioinformatics analysis shows that lncRNA-PVT1 contains one conserved target binding site of miR-1207-3p, the predicted complementary binding sites between lncRNA-PVT1 and miR-1207-3p. (D) miR-1207-3p expression in multiple solid tumors and normal tissues from TCGA Pan-Cancer dataset. (E) Dual-luciferase assay showed miR-1207-3p mimics reduced the luciferase activity of lncRNA-PVT1-Wt. (F) Low miR-1207-3p expression was associated with poor OS and DFS in patients with glioma. **p<0.001.
Figure 5
Figure 5
HNF1B is the direct target gene of miR-1207-3p and associated with poor prognosis of glioma. (A) The predicted complementary binding sites between miR-1207-3p and HNF1B. (B) Dual-luciferase assay showed miR-1207-3p mimics reduced the luciferase activity of HNF1B-Wt. (C) miR-1207-3p mimics reduced HNF1B expression in glioma cells, while miR-1207-3p inhibitors increased HNF1B mRNA expression. (D) IHC assays showed HNF1B expression was upregulated and associated advanced TNM stage in patients with glioma. (E, F) Western blot results showed overexpression of miR-489 significantly down-regulated HNF1B in U87 and U251 cells. *p<0.05.
Figure 6
Figure 6
lncRNA-PVT1 inhibition reduced tumor growth in vivo, and lncRNA-PVT1/miR-1207-3p/HNF1B axis promoted glioma metastasis. (A) Representative images of tumors in xenograft mouse model. (B)The relative weights of xenograft tumors. (C) The growth curves of xenograft tumors. (D, E) HALLMARK and KEGG Pathway Enrichment in glioma from microarry datasets. The datasets were analyzed using the Hallmark gene signature collection. Hallmark_EMT was identified as the pathway with the top two-highest association. (F) The expressions of HNF1B, EMT-related makers and VEGF-related molecules were determined using western blot. Proteins were isolated from cells transfected as indicated. *p<0.05, **p< 0.01, ***p<0.001.
Figure 7
Figure 7
Schema diagram of lncRNA-PVT1-miR-1207-3p/HNF1B axis in human glioma.
See this image and copyright information in PMC

References

    1. Reifenberger G, Wirsching HG, Knobbe-Thomsen CB, Weller M. Advances in the molecular genetics of gliomas - implications for classification and therapy. Nat Rev Clin Oncol. 2017;14(7):434–452. - PubMed
    1. Bi J, Chowdhry S, Wu S, Zhang W, Masui K, Mischel PS. Altered cellular metabolism in gliomas - an emerging landscape of actionable co-dependency targets. Nat Rev Cancer. 2020 Jan;20(1):57–70. - PubMed
    1. Chen R, Smith-Cohn M, Cohen AL, Colman H. Glioma Subclassifications and Their Clinical Significance. Neurotherapeutics. 2017 Apr;14(2):284–297. - PMC - PubMed
    1. Gusyatiner O, Hegi ME. Glioma epigenetics: From subclassification to novel treatment options. Semin Cancer Biol. 2018 Aug;51:50–58. - PubMed
    1. Hervey-Jumper SL, Berger MS. Insular glioma surgery: an evolution of thought and practice. J Neurosurg. 2019 Jan 1;130(1):9–16. - PubMed