Fig. 7: Dual targeting of HSP70 and AURKA enhances N-Myc degradation and improves treatment in NEPC. | Nature Communications

Fig. 7: Dual targeting of HSP70 and AURKA enhances N-Myc degradation and improves treatment in NEPC.

From: Proteostasis perturbation of N-Myc leveraging HSP70 mediated protein turnover improves treatment of neuroendocrine prostate cancer

Fig. 7

a Cell proliferation was measured in CWR22Rv1 cells transfected with siRNA against negative control, HSP70, AURKA, or in combination for 5 days (n = 3 samples). b, c CWR22Rv1 cells were transfected with siRNA targeting AURKA or negative control and then treated with JG231. Cell proliferation (b) and N-Myc expression (c) were determined by cell counting or western blotting, respectively (n = 3 samples). d CWR22Rv1, H660, and UCDCaP-CR cells were treated with increasing doses of alisertib for 3 days, and the viable cells were counted. e The expression of N-Myc was detected in CWR22Rv1 cells treated with alisertib alone or combined with JG231. f CWR22Rv1, H660, and UCDCaP-CR cells were treated with alisertib (0.025 µM for CWR22Rv1, 0.1 µM for other cell lines), JG231 (0.1 µM) or the combination for 7-10 days, and the viable cells were counted (n = 3 samples). g CWR22Rv1 cells were treated with JG231 alone or with alisertib in the clonogenic assay. h Organoids from LuCaP93 PDX were treated with JG231 (0.1 µM), alisertib (0.25 µM) alone or in combination for 7 days. Cell viability was assayed by the CellTiter-Glo Luminescent assay and the live-and-dead cells were visualized by immunofluorescence (n = 3 samples). Scale bar represents 100 microns. i Mice bearing H660 xenografts were treated with vehicle control, JG231 (2 mg/Kg i.p), alisertib (10 mg/Kg p.o), or JG231 plus alisertib for 30 days (n = 6). Tumor volumes were measured twice weekly. Tumors were photographed and weighed. j, k Drug toxicity tests were conducted by measuring the blood urea nitrogen (BUN) and alanine aminotransferase (ALT) levels in the serum samples collected from animals in each group (n = 4 samples). l IHC staining of N-Myc and Ki67 in each group was performed. Scale bar represents 20 microns. For (a, f, and h–k), statistical significance was determined by one-way ANOVA with a Tukey multiple-comparison test. For (b), statistical significance was determined by an unpaired two-sided t-test. Results are the mean of three independent experiments ( ± S.D.). Source data are provided as a Source Data file.

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