Alpha-2 Heremans Schmid Glycoprotein (AHSG) promotes the proliferation of bladder cancer cells by regulating the TGF-β signalling pathway

Figures & data

Figure 1. Levels of Alpha-2 Heremans Schmid Glycoprotein (AHSG) in bladder cancer (BC) cells and tissues.

Notes: (A) The levels of AHSG in human normal bladder epithelial cell line SV-40-immortalized human uroepithelial cell line and BC cell line T24 and 5637 was detected by western blotting. *** P < 0.001, **** P < 0.0001. (B a) Levels of AHSG in adjacent tissues detected by immunohistochemistry, –, 400×, Scale bars, 100 µm; (B b) Levels of AHSG in BC tissue detected by immunohistochemistry, 1+, 400×, Scale bars, 100 µm; (B c) Immunohistochemical detection of the expression of AHSG in BC tissue, 2+, 400×, Scale bars, 100 µm; (B d) Immunohistochemistry detection of AHSG levels in BC tissue, 3+, 400×, Scale bars, 100 µm; (B e) AHSG levels in adjacent tissues and BC tissues detected by immunohistochemistry, ** P < 0.01.

Figure 2. Transfection efficiency of overexpressed/knockdown Alpha-2 Heremans Schmid Glycoprotein (AHSG) in T24 and 5637 detected by western blotting.

Notes: (A) AHSG overexpression/knockdown efficiency in T24 compared with the negative control, **** P < 0.0001; (B) AHSG overexpression/knockdown efficiency in 5637 compared with the negative control, *** P < 0.001, **** P < 0.0001.

Figure 3. Effects of overexpression/knockdown of Alpha-2 Heremans Schmid Glycoprotein (AHSG) on the proliferation and cell cycle of T24 and 5637.

Notes: (A-B) The proliferation ability and clone formation ability of T24 and 5637 detected by the CCK8 and clone formation assay after AHSG overexpression, * P < 0.05; (C-D) CCK8 and clone formation assays were performed to detect the changes in the proliferation and clone formation abilities of T24 and 5637 after AHSG knockdown, ** P < 0.01; (E-F) Flow cytometry for the detection of the changes in the cell cycle in T24 and 5637 after AHSG overexpression, * P < 0.05; (G-H) Flow cytometry for the detection of the changes in the cell cycle in T24 and 5637 after AHSG knockdown, ** P < 0.01.

Figure 3. (continued)

Figure 3. (continued)

Figure 3. (continued)

Figure 4. The effect of overexpression/knockdown of Alpha-2 Heremans Schmid Glycoprotein (AHSG) in T24 and 5637 on the phosphorylation level of Smad2/3 protein, a key molecule downstream of the TGF-β conventional pathway.

Notes: (A-B) After the overexpression of AHSG in T24 and 5637, the cells were induced by human recombinant TGF-β for 30–60 min, and the change in Smad2/3 protein phosphorylation levels (p-Smad2/3/total Smad2/3) was detected by western blotting, *** P < 0.001. (C-D) After AHSG knockdown in T24 and 5637, the cells were induced by human recombinant TGF-β for 30–60 min, and the change in Smad2/3 protein phosphorylation levels (p-Smad2/3/total Smad2/3) was detected by western blotting, * P < 0.05, **** P < 0.0001.

Figure 5. The levels of Alpha-2 Heremans Schmid Glycoprotein (AHSG) in the urine of bladder cancer (BC) patients.

Notes: (A) The levels of AHSG in the urine of BC patients were detected by enzyme-linked immunosorbent assay, * P < 0.05, **** P < 0.0001. (B) Receiver operating characteristic curve analysis of 64 BC patients and 64 healthy subjects, and the area under the curve value is 0.9548 (95%CI: 0.9188 to 0.9908,**** P < 0.0001).