lncRNA CRNDE is Upregulated in Glioblastoma Multiforme and Facilitates Cancer Progression Through Targeting miR-337-3p and ELMOD2 Axis

Abstract

Introduction: Colorectal neoplasia differentially expressed (CRNDE) was reported to promote carcinogenesis in several cancers. However, the role of CRNDE in glioblastoma multiforme (GBM) needs to be further explored.

Methods: CRNDE expression levels in GBM tissues and cells were explored using real-time quantitative PCR at first. Effects of CRNDE on GBM cell behaviors were detected by conducting in vitro experiments. Interactions of CRNDE, microRNA-337-3p (miR-337-3p), and ELMO domain containing 2 (ELMOD2) were verified by bioinformatics analysis tools and dual-luciferase reporter assay. Expression correlations of CRNDE and ELMOD2 in GBM tissues were analyzed at GEPIA website.

Results: CRNDE expression was upregulated in GBM tissues and cells compared with normal counterparts. CRDNE knockdown inhibits proliferation and migration, but promotes apoptosis in GBM cell, while CRNDE overexpression caused opposite effects. Mechanisms exploration indicated CRNDE serves as sponge of miR-337-3p to upregulate ELMOD2 expression. Furthermore, we showed CRNDE and ELMOD2 were positively correlated in GBM tissues.

Discussion: In conclusion, our study highlighted the importance of CRNDE/miR-337-3p/ELMOD2 axis in GBM progression and offered novel strategies for GBM treatment.

Keywords: CRNDE; ELMOD2; glioblastoma multiforme; miR-337-3p.

Figure 1

Highly expressed of CRNDE in GBM. (A) CRNDE expression in GBM tissues and normal tissues detected at GEPIA. (B) CRNDE expression in GBM tissues and normal tissues detected at Oncomine. (C) CRNDE expression in GBM cells and normal cell. (D) Correlation of CRNDE expression and overall survival of GBM patients. ***P < 0.001; *P < 0.05.

Figure 2

CRNDE regulates GBM cell growth, migration, and invasion. (A) Transfection efficiency of si-CRNDE and pCRNDE in GBM cell. (B) Cell proliferation rate of GBM cells transfected with si-CRNDE or pCRNDE. (C) Cell apoptosis percentage of GBM cells transfected with si-CRNDE or pCRNDE. (D) Cell migration ability of GBM cells transfected with si-CRNDE or pCRNDE. (E) Cell invasion ability of GBM cells transfected with si-CRNDE or pCRNDE. ***P < 0.001, **P < 0.01.

Figure 3

CRNDE serves as sponge for miR-337-3p in GBM. (A) Prediction result between CRNDE and miR-337-3p. (B) miR-337-3p expression in GBM cells and normal cell. (C) Luciferase activity reporter assay showed direct interaction between CRNDE and miR-337-3p. (D) Cell proliferation rate of GBM cells transfected with miR-scramble+pcDNA3.1, miR-scramble+pCRNDE, miR-337 mimic+pcDNA3.1, or miR-337 mimic+pCRNDE. (E) Cell apoptosis percentage of GBM cells transfected with miR-scramble+pcDNA3.1, miR-scramble+pCRNDE, miR-337 mimic+pcDNA3.1, or miR-337 mimic+pCRNDE. (F) Cell migration ability of GBM cells transfected with miR-scramble+pcDNA3.1, miR-scramble+pCRNDE, miR-337 mimic+pcDNA3.1, or miR-337 mimic+pCRNDE. (G) Cell invasion ability of GBM cells transfected with miR-scramble+pcDNA3.1, miR-scramble+pCRNDE, miR-337 mimic+pcDNA3.1, or miR-337 mimic+pCRNDE. ***P < 0.001, **P < 0.01, *P < 0.05.

Figure 4

ELMOD2 serves as target for miR-337-3p. (A) Prediction result between ELMOD2 and miR-337-3p. (B) ELMOD2 expression in GBM tissues and normal tissues detected at GEPIA. (C) ELMOD2 expression in GBM tissues and normal tissues detected at Oncomine. (D) ELMOD2 expression in different tumor grades detected at Oncomine. (E) ELMOD2 expression in GBM cells and normal cell detected by RT-qPCR. (F) ELMOD2 expression in GBM cells and normal cell detected by Western blot. (G) Luciferase activity reporter assay showed direct interaction between ELMOD2 and miR-337-3p. ***P < 0.001, **P < 0.01, *P < 0.05.

Figure 5

CRNDE modulates ELMOD2 by binding with miR-337-3p. (A) miR-337-3p and ELMOD2 expression in GBM cells with pCRNDE transfection. (B) ELMOD2 expression in GBM cells with pcDNA3.1+miR-Scramble, pcDNA3.1+siR-Scramble, pCRNDE+miR-Scramble, pCRNDE+miR-337-3p mimic, or pCRNDE+si-ELMOD2 transfection detected by RT-qPCR. (C) ELMOD2 expression in GBM cells with pcDNA3.1+miR-Scramble, pcDNA3.1+siR-Scramble, pCRNDE+miR-Scramble, pCRNDE+miR-337-3p mimic, or pCRNDE+si-ELMOD2 transfection detected by Western blot. (D) Correlation between CRNDE and ELMOD2 in GBM tissues. (E) Cell proliferation rate of GBM cells transfected with pcDNA3.1+siR-Scramble, pcDNA3.1+siR-Scramble, pCRNDE+siR-Scramble, pCRNDE+miR-337-3p mimic, or pCRNDE+si-ELMOD2. (F) Cell apoptosis percentage of GBM cells transfected with pcDNA3.1+miR-Scramble, pcDNA3.1+siR-Scramble, pCRNDE+miR-Scramble, pCRNDE+miR-337-3p mimic, or pCRNDE+si-ELMOD2. (G) Cell migration ability of GBM cells transfected with pcDNA3.1+miR-Scramble, pcDNA3.1+siR-Scramble, pCRNDE+miR-Scramble, pCRNDE+miR-337-3p mimic, or pCRNDE+si-ELMOD2. (H) Cell invasion ability of GBM cells transfected with pcDNA3.1+miR-Scramble, pcDNA3.1+siR-Scramble, pCRNDE+miR-Scramble, pCRNDE+miR-337-3p mimic, or pCRNDE+si-ELMOD2. ***P < 0.001, **P < 0.01, *P < 0.05.

Figure 6

CRNDE promotes GBM tumorigenesis in vivo. (A) Tumor size, (B) Tumor volume, and (C) Tumor weight in GBM cells with CRNDE knockdown or not were measured in vivo. (D) RT-qPCR was used to detect CRNDE, miR-337-3, and ELMOD2 expression in animal model with CRNDE knockdown. (E) Survival rate of nude mice bearing sh-CRNDE or shR-scramble. ***P < 0.001, **P < 0.01.