Effects of PAFAH1B3 on proliferation, migration, and aerobic glycolysis of bladder cancer cells via regulating CRYAB/PI3K/Akt pathway
10.3760/cma.j.cn121382-20250703-00051
- VernacularTitle:PAFAH1B3调控CRYAB/PI3K/Akt通路促进膀胱癌细胞的增殖、迁移和有氧糖酵解
- Author:
Chenyun LI
1
;
Na ZHUO
;
Tong SUN
;
Jing LI
Author Information
1. 天津医科大学第二医院超声科，天津 300211
- Keywords:
Bladder cancer;
Platelet activating factor acetylhydrolase 1b catalytic subunit 3;
αB-crystallin;
Phosphoinositide 3-kinase;
Protein kinase B;
Aerobic glyc
- From:
International Journal of Biomedical Engineering
2025;48(5):454-461
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expression of platelet activating factor acetylhydrolase 1b catalytic subunit 3 (PAFAH1B3) in bladder cancer cells, and to explore the effects of PAFAH1B3 on proliferation, migration, and aerobic glycolysis in bladder cancer T24 cells and its mechanism.Methods:The relative expression of PAFAH1B3 in bladder cancer RT4, 5637, T24, J82T24 cells and human normal bladder epithelial SV-HUC-1 cells were detected using Western blotting. After culture, T24 cells were divided into a control group and a knockdown group based on treatment conditions. T24 cells were transfected with 50 nmol/L of negative control small interfering RNA (siRNA) or PAFAH1B3 siRNA, respectively. T24 cells with PAFAH1B3 knockdown were subsequently infected with an adenovirus vector overexpressing αB-crystallin ( CRYAB) at a multiplicity of infection of 50, and were designated as the overexpression group. The relative expression of PAFAH1B3 in T24 cells was detected using Western blotting. The cell survival rate, the number of colonies per unit field of view and the scratch closure rate of T24 cells were assessed using cell counting kit-8, colony formation assay and scratch assay, respectively. Glucose uptake, lactate production and adenosine triphosphate (ATP) levels of T24 cells were measured with corresponding commercial kits. The relative expression levels of glucose transporter 1 (GLUT1), hexokinase 2 (HK2), lactate dehydrogenase A (LDHA), CRYAB, phosphorylated phosphoinositide 3-kinase (p-PI3K) to PI3K and phosphorylated protein kinase B (p-Akt) to Akt were detected using Western blotting. Data were analyzed by an independent sample t test or one-way analysis of variance. Results:The relative expression levels of PAFAH1B3 protein in RT4, 5637, T24 and J82 cells (1.13±0.15, 1.40±0.10, 1.50±0.10, 0.77±0.13) were significantly higher than that in SV-HUC-1 cells (0.42±0.08) (all P<0.01). The relative expression levels of PAFAH1B3 protein in the knockdown group (0.22±0.08) was significantly lower than that in the control group (1.08±0.03) ( P<0.01). The cell survival rate, the number of colonies per unit field of view and the scratch closure rate in the knockdown group [(54.00±6.00)%, (104.00±8.00) cells, (42.00±3.00)%] were all lower than those in the control group [(96.00±3.00)%, (170.00±6.00) cells, (72.00±4.00)%] (all P<0.01). Glucose uptake, lactate production and ATP level in the knockdown group [(0.46±0.09) mg/ml, (22.67±2.67, 7.50±0.15) μmol/L] were lower than those in the control group [(1.21±0.11) mg/ml, (41.67±2.33, 18.00±2.00) μmol/L] (all P<0.01). The relative expression levels of GLUT1, HK2 and LDHA proteins in the knockdown group (0.31±0.04, 0.18±0.03, 0.47±0.07) were all lower than those in the control group (0.89±0.09, 0.97±0.04, 0.95±0.03) (all P<0.01). The relative expression of CRYAB, p-PI3K/PI3K and p-Akt/Akt proteins in the knockdown group (0.26±0.04, 0.44±0.03, 0.31±0.04) were all lower than those in the control group (0.99±0.05, 0.94±0.05, 0.99±0.06) (all P<0.01). The relative expression of CRYAB in the overexpression group (4.30±0.40) was higher than that in the knockdown group (0.94±0.05) ( P<0.01). The cell survival rate and the number of colonies per unit field of view in the overexpression group [(92.00±3.00)%, (172.00±8.00) cells] were all higher than those in the knockdown group [(46.00±3.00)%, (103.00±7.00) cells] (both P<0.01). Glucose uptake, lactate production and ATP level in the overexpression group [(1.25±0.06) mg/ml, (43.00±4.00, 21.00±2.00) μmol/L] were all higher than those in the knockdown group [(0.49±0.06) mg/ml, (19.00±3.00, 7.00±1.00) μmol/L] (all P<0.01). The relative expression levels of GLUT1, HK2 and LDHA proteins in the overexpression group (0.71±0.04, 0.98±0.04, 0.99±0.04) were higher than those in the knockdown group (0.26±0.03, 0.52±0.03, 0.21±0.02) (all P<0.01). The relative expression levels of p-PI3K/PI3K and p-Akt/Akt proteins in the overexpression group (0.77±0.03, 0.96±0.04) were all higher than those in the knockdown group (0.24±0.05, 0.18±0.03) (both P<0.01). Conclusions:PAFAH1B3 may promote the proliferation, migration and aerobic glycolysis of bladder cancer cells via regulation of the CRYAB/PI3K/Akt signaling pathway.
