SNHG4 was upregulated in glioma tissues and cell lines and ssociated with prognosis of patients. a The expression level of SNHG4 in multiple cancer tissues was analyzed by UALCAN website comparing to normal tissues, including glioma tissues. b The correlation between overall sruvival of glioma patients and high SNHG4 expression was analyzed by Kaplan–Meier plotter. c. The expression level of SNHG4 was measured by qRT-PCR in human glioma cell lines (Ln229, U251,U87) compared with the normal brain astrocyte cell line (SVG) (P = 0.0090 and 0.0099). **: P < 0.01

SNHG4 promoted the proliferation and migration of glioma cells in vitro. a The knockdown efficiency of SNHG4 was detected in Ln229 cells by qRT-PCR assay when transfection with si1-SNHG4, si2-SNHG4 and NC (P = 0.0027 and 0.0002). b The knockdown efficiency of SNHG4 was detected in U251 cells by qRT-PCR assay when transfection with si1-SNHG4, si2-SNHG4 and NC (P = 0.0030 and 0.0011). c. The overexpression efficiency of SNHG4 was detected in U87 cells by qRT-PCR assay when transfection with pcDNA3.1-SNHG4 plasmids (P = 0.0492). d, e CCK-8 assay were used to assess the proliferation of Ln229 (D) and U251 (E) cells after knocking-down SNHG4. f For overexpression of SNHG4 in U87 cells, the CCK-8 assay was used to assess the cell proliferation. g, h Transwell assays were performed to examine cell migration in Ln229 (g) and U251 (h) cells transfected with SNHG4 siRNAs (P = 0.0156 and 0.0064, P = 0.0067 and 0.0101). i. Transwell assays were performed to examine cell migration in U87 cells when transfection with pcDNA3.1-SNHG4 plasmids. *: P < 0.05, **: P < 0.01, ***: P < 0.001

Knockdown of SNHG4 decreases the growth and metastasis of glioma cells in zebrafish xenograft. a The glioma cells transfected with si-SNHG4 or NC were injected into the PVS of 2-dpf (days post‑fertilization) wild type zebrafish larvae. Images of the yolk were taken by a stereomicroscope at 4 dpi. b The CM-DiI-positive areas in the yolk were quantified for cell proliferation. c The typical images were imaged by confocal microscope. d The glioma cells transfected with si-SNHG4 or NC were injected into the PVS of 2-dpf wild type zebrafish larvae. Images of the trunk were taken by a stereomicroscope at 4 dpi. e The CM-DiI-positive areas in the trunk were quantified for cell metastasis. f The typical images were imaged by confocal microscope. g, h Statistical analysis of proliferation (g) and metastasis (h) when knocking down SNHG4 in Ln229 cells (n = 17), compared with NC (n = 16) (P = 0.0001 and 0.0131). i, j Statistical analysis of proliferation (i) and metastasis (j) when knocking down SNHG4 in U251 cells (n = 14), compared with NC (n = 11) (P = 0.0002). *: P < 0.05, ***: P < 0.001. Scale: 100 μm

SNHG4 was a ceRNA to sponge miR-367-3p. a Subcellular localization of SNHG4 was analyzed by LncLocator website. b The potential miRNAs which might bind to SNHG4 by StarBase. c, d The expression levels of six miRNAs were examined when knockdown of SNHG4 in Ln229 (c) and U251 (d) cells by qRT-PCR (P < 0.0002 and 0.0249). e The potential binding site between miR-367-3p and SNHG4 was predicted by StarBase. f Schematic diagram of luciferase reporter plasmids construction, named pGL3-promoter-SNHG4-WT and pGL3-promoter-SNHG4-MUT. g Luciferase reporter assay was performed to assess the interaction between LINC01123 and miR-384 (P = 0.0031). *: P < 0.05, **: P < 0.01, ***: P < 0.01

SNHG4 interacts with miR‐367-3p to regulate glioma cell proliferation and migration. a, b The overexpression efficiency of miR-367-3p in Ln229 (a) and U251 (b) cells when transfecting its mimics by qRT-PCR assay (P = 0.0106 and 0.0234). c, d The inhibitory efficiency of miR-367-3p in Ln229 (c) and U251 (d) cells when transfecting its inhibitor by qRT-PCR assay (P = 0.0163 and 0.0313). e, f The proliferation was assessed by CCK-8 assay when overexpression miR-367-3p in Ln229 (e) and U251 (f) cells. g, h The migration was assessed by Transwell assay when overexpression miR-367-3p in Ln229 (g) and U251 (h) cells (P = 0.0486). i, i. CCK-8 assay was used to evaluate cell proliferation and migration when co-transfection of miR-384 inhibitor and si-SNHG4 in Ln229 (i) and U251 (j) cells. k, l Transwell assays was used to evaluate cell migration when co-transfection of miR-384 inhibitor and si-SNHG4 in Ln229 (k) and U251 (l) cells (P = 0.0207 and 0.0022, 0.0029 and 0.0011). *: P < 0.05, **: P < 0.01, ***: P < 0.01

MYO1B is a target of miR-367-3p, and miR-367-3p regulates the progression of glioma through MYO1B. a The target genes of miR-367-3p was predicted by miRDB website. b, c The expression levels of target genes were detected in mRNA levels by qRT-PCR assay after overexpression of miR-367-3p in Ln229 (b) and U251 (c) cells (P = 0.0128 and 0.0062, 0.0002 and 0.0072). d The expression levels of MYO1B were detected in protein levels by western blot assay after overexpression of miR-367-3p in Ln229 and U251 cells. e The potential binding site between miR-367-3p and MYO1B 3’UTR was predicted by StarBase. f Schematic diagram of luciferase reporter plasmids construction, named pGL3-promoter-MYO1B-WT and pGL3-promoter-MYO1B-MUT. g Luciferase reporter assay was performed to assess the interaction between MYO1B 3’UTR and miR-367-3p (P = 0.0004). h, i The overexpression efficiency of MYO1B when transfection with pcDNA3.1- MYO1B plasmids in Ln229 (h) and U251 (i) cells at mRNA levels by qRT-PCR assays (P = 0.0039 and 0.0212). j. The overexpression efficiency of MYO1B when transfection with pcDNA3.1- MYO1B plasmids in Ln229 and U251 cells at protein level by western blot assays (P = 0.0039 and 0.0212). k, l CCK-8 assay was used to evaluate cell proliferation and migration when co-transfection of miR-367-3p mimics and pcDNA3.1- MYO1B in Ln229 (k) and U251 (l) cells. m, n Transwell assays were used to evaluate cell migration when co-transfection of miR-367-3p mimics and pcDNA3.1- MYO1B in Ln229 (m) and U251 (n) cells (P = 0.0495 and 0.0034). *: P < 0.05, **: P < 0.01, ***: P < 0.01

SNHG4 regulated the proliferation and migration of glioma cells via MYO1B. a, b The expression levels of MYO1B were examined after silencing SNHG4 in Ln229 (a) and U251 (b) cells at mRNA level by qRT-PCR assay (P = 0.0042 and 0.0001, 0.0126 and 0.0362). c, d The expression levels of MYO1B were examined after silencing SNHG4 in Ln229 (c) and U251 (d) cells at protein level by western blot assays. e, f CCK-8 assay was used to evaluate cell proliferation when co-transfection of si-SNHG4 and pcDNA3.1-MYO1B plasmid in Ln229 (e) and U251 (f) cells. g, h Transwell assay was used to evaluate cell migration when co-transfection of si-SNHG4 and pcDNA3.1-MYO1B plasmid in Ln229 (g) and U251 (h) cells (P = 0.0004 and 0.0012, 0.0004 and 0.0017). *: P < 0.05, **: P < 0.01, ***: P < 0.001