Figure 9.

SR3029 represses the growth of APC^min/+ colorectal tumor organoids and patient-derived colorectal tumor xenografts (PDTX) through enhancing AES expression. (A) Morphology of colorectal tumor organoids from B6-APC^min/+ mice treated with SR3029. The organoids were treated with the indicated amounts of SR3029 for 24 h. Then SR3029 was removed and the organoids were grown in normal medium for another 5 days before photomicrographed. Right panel: graphical representation of quantitative data showed the relative size of organoid. Scale bar, 200 μm. (B) AES staining of colorectal tumor organoids treated with or without SR3029. Scale bar, 50 μm. (C) Representative images of tumors from the control and SR3029-treated PDTX. (D) Mean tumor volume (n = 10). (E) Mean tumor weight (n = 10). (F) Total RNA was extracted from tumor samples treated with or without SR3029 and real-time PCR was performed to detect the mRNA expression of Wnt target genes (Axin2, Fibronectin and LEF1), Wnt-related stemness marker genes (CD44 and LGR5) and Notch target genes (HES1 and HES2). Quantification of mRNA level was normalized to GAPDH (n = 5). (G) H&E, Ki-67, AES, CD44, and LGR5 staining of control and SR3029-treated PDTX. Scale bar, 50 μm. (H) The expression levels of LEF1, CD44, LGR5, PCNA and AES in tumor samples from the control and SR3029-treated PDTX were detected by immunoblotting. Values are shown as means ± SD. *P < 0.05; Two-way ANOVA for (D); Student's t test for (A), (E) and (F).