Fig. 2. PTHrP increases cell viability and tumorsphere formation of patient-derived GSCs. (A∼C) Three patient-derived GSCs (GSC-01, GSC-02, and GSC-03) were treated with a series of recombinant human PTHrP proteins (rPTHrP; 1, 3, 5, 10, 50, and 100 nM/ml) for 3 days, and cell viability was detected by CCK‐8 assay. *p<0.05, **p<0.01, ***p<0.001 versus control (Ctrl) group. (D) Patient-derived GSCs were transfected with FAM-labeled non-specific siRNAs (siNC) or PTHrP-targeted siRNA (siPTHrP). At the next day, more than 90% of transfected cells were observed. Scale bar=400 μm. (E, F) The expression of PTHrP was detected by western blot. Data from three independent experiments (n=3) were presented as the ratio of PTHrP to β‐Actin. ***p<0.001 versus the siNC group. After transfecting with siNC or siPTHrP, the tumorsphere number (G∼I) and the mean diameter (J∼L) were measured at 3 days, 5 days, and 7 days. Data were represented as the mean±SD of three independent experiments (n=3). (M, N) The PTHrP expression wsa examined by western blot. Data from three independent experiments (n=3) were presented as the ratio of PTHrP to β‐Actin. ***p<0.001 versus shNC group. The luciferase-expressing shNC GSCs or KD-PTHrP GSCs were used to establish the orthotopic glioma mouse model. (O, P) The in vivo imaging of shNC GSCs and KD-PTHrP GSCs in tumor-bearing mice at different time points after injection. Data are presented as the mean±standard deviation of five independent experiments (n=5). *p<0.05, **p<0.01 versus the shNC group.

International Journal of Stem Cells 2023;16:315-25 https://doi.org/10.15283/ijsc22097

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