Fig. 3.

Inhibition of PIK3C3 in miR-338-5p stable cells promoted CRC metastasis in vivo. HCT116 stable cells were injected into the spleens of NOD and SCID mice. After 42 days, the spleen, liver, and lung were dissected out. (a) Kaplan-Meier analysis was used to calculate the overall survival rate of mice (P values were analysed using Log Rank tests). (b) The amount of ascites was measured and tumour cells in ascites were demonstrated by Liu stain. Data are presented as median (IQR) (n = 5) (P values were analysed using Mann-Whitney tests). (c) Tumours in the spleen, liver, and lung were examined by hematoxylin and eosin (H&E) stain and PIK3C3 IHC staining (100×). The metastatic tumours were indicated by arrow point. IHC showed that PIK3C3 was high expression in primary tumour and metastatic nodules, when PIK3C3 was overexpressed. (d) The number and volume of metastatic tumours were measured in the liver and lung, respectively. The scale bar = 200 μm. Data are presented as median (IQR) (n = 5) (P values were analysed using Mann-Whitney tests). (e) Expression of miR-338-5p or PIK3C3 RNA in primary tumour of spleen was assayed by QPCR. Data are expressed as median (IQR) (n = 5) (P values were analysed using Mann-Whitney tests). IHC of PIK3C3 expression (f) and ratio of miR-338-5p/PIK3C3 (g) were measured in the splenic primary tumour, metastasis in the liver and lung, respectively. Data are expressed as median (IQR) (n = 5) (P values were analysed using Mann-Whitney tests). Correlation was calculated using linear regression (Data were analysed by Spearman tests).