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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

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. 5

CircATP5B promoted the proliferation of GSCs by upregulating HOXB5 expression. a, b, c qRT-PCR (a) and western blotting (b, c) showed the expression of HOXB5 in GSCs after circATP5B knockdown or overexpression. (GSC406: p < 0.01; GSC201: p < 0.001; One-Way ANOVA). d The decreased expression of HOXB5 in GSC406 induced by circATP5B knockdown was reversed by miR-185-5p inhibitor treatment, as detected by western blotting. e The increased expression of HOXB5 in GSC201 induced by circATP5B overexpression was reversed by miR-185-5p mimic treatment, as determined by western blotting. f The effects of both circATP5B and miR-185-5p on the expression of HOXB5 in GSCs were detected by qRT-PCR. (GSC406: p < 0.01; GSC201: p < 0.01; One-Way ANOVA). g The relative expression correlation between circATP5B and HOXB5 in 70 cases of glioma patients were detected by qRT-PCR. (Total: r = 0.4625, p = 0.04; Grade II: r = 0.4840, p = 0.0142; Grade III: r = 0.5333, p = 0.006; Grade IV: r = 0.6290, p < 0.0001; Pearson’s correlation analyses). h and i MTS assays showed that circATP5B knockdown or overexpression affected the cell viability of GSCs and was reversed by HOXB5 overexpression or knockdown, respectively. (GSC406: p < 0.001; GSC201: p < 0.001; One-Way ANOVA). j The EDU assays showed that circATP5B knockdown or overexpression affected the proliferation of GSCs and was reversed by HOXB5 overexpression or knockdown, respectively. Scale bar = 100 μm. (GSC406: p < 0.01; GSC201: p < 0.01; One-Way ANOVA). k The neurospheres formation assays showed that circATP5B knockdown or overexpression affected the relative size of the neurospheres of GSCs and was reversed by HOXB5 overexpression or knockdown, respectively. Scale bar = 20 μm. (GSC406: p < 0.01; GSC201: p < 0.01; One-Way ANOVA). l and m Limiting dilution assays showed that circATP5B knockdown or overexpression affected the neurosphere-forming capacity of GSCs and was reversed by HOXB5 overexpression or knockdown, respectively (GSC406: p < 0.01; 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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