Lactobacillus fermentum ZS09 Mediates Epithelial–Mesenchymal Transition (EMT) by Regulating the Transcriptional Activity of the Wnt/β-Catenin Signalling Pathway to Inhibit Colon Cancer Activity

Figures & data

Figure 1 Gram staining microscopic examination. (A) Lactobacillus fermentum ZS09; (B) Lactobacillus bulgaricus.

Figure 2 (A) Colon tissue morphology. The arrow shows the location of the colon mass. (B) Mouse survival curve.

Abbreviations: NC, normal untreated mice; CRC, colon cancer model mouse induced with AOM-DSS; ZS09-L, mouse treated with low-dose L. fermentum ZS09 (10⁹ CFU); ZS40-H, mouse treated with high-dose L. fermentum ZS09 (10¹¹ CFU); BLA, mouse treated with L. bulgaricus strain (10¹¹ CFU); SD, mouse treated with sulfasalazine (2.5%).

Figure 3 Mouse serum indicators.

Abbreviations: MMP-9, matrix metallopeptidase 9; TNF-α, tumour necrosis factor; IL-6R, interleukin 6; Ang-2, angiopoietin 2; VEGFR-2, vascular endothelial growth factor receptor 2; DKK1, Dickkopf 1; NC, normal untreated mice; CRC, colon cancer model mouse induced with AOM-DSS; ZS09-L, mouse treated with low-dose L. fermentum ZS09 (10⁹ CFU); ZS40-H, mouse treated with high-dose L. fermentum ZS09 (10¹¹ CFU); BLA, mouse treated with L. bulgaricus strain (10¹¹ CFU); SD, mouse treated with sulfasalazine (2.5%).

Figure 4 (A) Pathological section of mouse colon (H&E). Magnification 100×. (B) Colon severity pathological scores. 0 marks: intact villi and epithelium; 1 marks: slight submucosal or lamina propria swelling and separation; 2 marks: moderate submucosa or lamina propria swelling and separation and plasma cell infiltration; 3 marks: severe submucosal or lamina propria swelling and plasma cell infiltration, local villi atrophy and shedding; 4 marks: intestinal villi disappeared with intestinal wall necrosis. Pathology score ≥ 2, regarded as intestinal injury.

Abbreviations: NC, normal untreated mice; CRC, colon cancer model mouse induced with AOM-DSS; ZS09-L, mouse treated with low-dose L. fermentum ZS09 (10⁹ CFU); ZS40-H, mouse treated with high-dose L. fermentum ZS09 (10¹¹ CFU); BLA, mouse treated with L. bulgaricus strain (10¹¹ CFU); SD, mouse treated with sulfasalazine (2.5%).

Figure 5 (A) Pathological section of mouse colon (CD34 immunohistochemical staining). Magnification 200×. (B) Quantitative immunohistochemical density. AOD (Average Optical Density) = IOD/ Area.

Abbreviations: NC, normal untreated mice; CRC, colon cancer model mouse induced with AOM-DSS; ZS09-L, mouse treated with low-dose L. fermentum ZS09 (10⁹ CFU); ZS40-H, mouse treated with high-dose L. fermentum ZS09 (10¹¹ CFU); BLA, mouse treated with L. bulgaricus strain (10¹¹ CFU); SD, mouse treated with sulfasalazine (2.5%).

Figure 6 (A) mRNA expression levels of inflammatory factors in colon tissue. *Represents a significant difference from the NC group. *p <0.05, **<0.01, ***<0.001, ****<0.0001 (B) Expression of key proteins in signaling pathway in colon tissue.

Abbreviations: NC, normal untreated mice; CRC, colon cancer model mouse induced with AOM-DSS; ZS09-L, mouse treated with low-dose L. fermentum ZS09 (10⁹ CFU); ZS40-H, mouse treated with high-dose L. fermentum ZS09 (10¹¹ CFU); BLA, mouse treated with L. bulgaricus strain (10¹¹ CFU); SD, mouse treated with sulfasalazine (2.5%).