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Fig. 1 | Journal of Experimental & Clinical Cancer Research

Fig. 1

From: The SRSF1/circATP5B/miR-185-5p/HOXB5 feedback loop regulates the proliferation of glioma stem cells via the IL6-mediated JAK2/STAT3 signaling pathway

Fig. 1

CircATP5B was upregulated in glioma and correlated with poor patient survival, and regulated the proliferation of GSCs. a CircATP5B was expressed at higher levels in glioma tissues, compared with adjacent brain tissues as detected by qRT-PCR. (Adjacent vs. Tumor: n = 10, p < 0.0001, Student’s t-test). b The expression level of circATP5B in different glioma tissues as detected by qRT-PCR. (grade II, n = 20; grade III, n = 25; grade IV, n = 25; III vs. II: p < 0.0001, IV vs. III: p < 0.0001, IV vs. II: p < 0.0001, One-Way ANOVA). c Schematic illustration of the formation of circATP5B via the circularization of exons in the ATP5B gene. d Sanger sequencing confirmed the head-to-tail splicing of circATP5B. e Relative expressions of circATP5B and ATP5B in both GSC406 and GSC201 were detected by qRT-PCR in the presence or absence of RNase R. (GSC201: p < 0.0001; GSC406: p < 0.0001; Student’s t-test). f circATP5B was mainly located in the cytoplasm by nuclear-cytoplasmic fractionation assay. (GSC201: U6: p < 0.0001, GAPDH: p = 0.026, circATP5B: p = 0.0057; GSC406: U6: p < 0.0001, GAPDH: p = 0.032, circATP5B: p = 0.0082, Student’s t-test). g Kaplan-Meier analysis showed the prognostic significance of the 70 glioma patients, 45 LGG patients, and 25 GBM patients with high versus low circATP5B expression detected by qRT-PCR. (LGG: p = 0.0126; GBM: p = 0.0092; 70 cases: p = 0.0017; Log-rank test). h MTS assays showed that circATP5B knockdown or overexpression affected the cell viability of GSCs (GSC406: p < 0.0001; GSC201: p < 0.0001; One-Way ANOVA). i The EDU assays showed that circATP5B knockdown or overexpression affected the proliferation of GSCs. Scale bar = 100 μm. (GSC406: p < 0.01; GSC201: p < 0.001; One-Way ANOVA). j The neurospheres formation assays showed that circATP5B knockdown or overexpression affected the relative size of the neurospheres of GSCs. Scale bar = 20 μm. (GSC406: p < 0.01; GSC201: p < 0.01; One-Way ANOVA). k circATP5B knockdown or overexpression affected the neurosphere-forming capacity of GSCs as detected by limiting dilution assays (GSC406: p < 0.05; GSC201: p < 0.05; ELDA analysis; circles represent corresponding points, triangles mean the point is outside of the log fraction number wells). EV: empty vector, OE: overexpression, NC: negative control, KD: knockdown. All data were expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001

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