1.PTBP1 promotes osteosarcoma cell proliferation and inhibits apoptosis via the PTEN-Akt pathway
Yikemu XIRENIJIANG ; Chengli DA ; Kamilijiang ROZI ; Abulizi TAJIGULI ; Mijiti REZIYE
Cancer Research and Clinic 2025;37(8):591-596
Objective:To investigate the effect of polypyrimidine tract binding protein 1 (PTBP1) on the proliferation and apoptosis of osteosarcoma cell lines via the phosphatase and tensin homolog (PTEN)-protein kinase B (Akt) signaling pathway.Methods:Osteosarcoma cell lines MG63, U2-OS, Saos-2, 143B, and the osteoblast cell line hFOB1.19 were selected. Western blotting was used to detect the relative expression level of PTBP1 protein, and the Saos-2 cell line with the highest PTBP1 relative expression level was selected for subsequent experiments. Saos-2 cells were divided into 5 groups: si-NC group [transfected with negative control siRNA (NC siRNA)], si-PTBP1 group (transfected with PTBP1 siRNA), solvent control+si-NC group [treated with dimethyl sulfoxide(DMSO) to a final concentration of 0.1% and transfected with NC siRNA], solvent control+si-PTBP1 group (treated with 0.1% DMSO and transfected with PTBP1 siRNA), and SF1670+si-PTBP1 group (treated with 10 μmol/L SF1670, a PTEN inhibitor, and transfected with PTBP1 siRNA). CCK-8 assay was used to detect the cell proliferation [expressed as the absorbance value at 490 nm ( A490)]; TUNEL staining was used to detect apoptosis ability, and apoptosis rate was calculated; Western blotting was used to detect the expression levels of relevant proteins. Results:The relative expression level of PTBP1 protein in osteosarcoma MG63, U2-OS, Saos-2, 143B, and osteoblast cell line hFOB1.19 cells was 0.80±0.12, 0.55±0.08, 1.04±0.10, 0.49±0.07, and 0.26±0.05, respectively, and the difference was statistically significant ( F = 47.10, P < 0.001). CCK-8 assay result showed that the A490 value of Saos-2 cells in si-NC group and si-PTBP1 group was 1.00±0.05 and 0.55±0.07, respectively, and the difference was statistically significant ( t = 10.46, P < 0.001). The result of TUNEL staining showed that the apoptosis rate was (1.26±0.05)% and (12.86±0.87)%, respectively in si-NC group and si-PTBP1 group, and the difference was statistically significant ( t = 26.62, P < 0.001). Western blotting showed that PTEN relative expression level of Saos-2 cell in si-PTBP1 group was higher than that in si-NC group (1.02±0.05 vs. 0.26±0.04, t = 23.74, P < 0.001), and the relative expression level of p-Akt in si-PTBP1 group was lower than that in si-NC group (0.22±0.03 vs. 1.00±0.07, t = 20.48, P < 0.001). In the solvent control+si-NC, solvent control+si-PTBP1, and SF1670+si-PTBP1 groups, the relative expression level of p-Akt protein was 0.96±0.07, 0.21±0.02, and 0.79±0.03, respectively, and the difference was statistically significant ( F = 299.30, P < 0.001), and the relative expression level of PTEN protein was 0.26±0.02, 0.98±0.08, and 0.96±0.11, respectively, and the difference was statistically significant ( F = 106.80, P < 0.001). The A490 values were 1.00±0.10, 0.53±0.08, and 0.89±0.08, respectively in the solvent control+si-NC, solvent control+si-PTBP1, and SF1670+si-PTBP1 groups, and the difference was statistically significant ( F = 31.81, P < 0.001); apoptosis rates were (1.27±0.11)%, (13.26±0.89)%, and (4.86±0.37)%, respectively, and the difference was statistically significant ( F = 482.90, P < 0.001). Conclusions:Knockdown of PTBP1 inhibits the proliferation and promotes the apoptosis of osteosarcoma cells, and this effect may be associated with the regulation of the PTEN-Akt pathway.
2.PTBP1 promotes osteosarcoma cell proliferation and inhibits apoptosis via the PTEN-Akt pathway
Yikemu XIRENIJIANG ; Chengli DA ; Kamilijiang ROZI ; Abulizi TAJIGULI ; Mijiti REZIYE
Cancer Research and Clinic 2025;37(8):591-596
Objective:To investigate the effect of polypyrimidine tract binding protein 1 (PTBP1) on the proliferation and apoptosis of osteosarcoma cell lines via the phosphatase and tensin homolog (PTEN)-protein kinase B (Akt) signaling pathway.Methods:Osteosarcoma cell lines MG63, U2-OS, Saos-2, 143B, and the osteoblast cell line hFOB1.19 were selected. Western blotting was used to detect the relative expression level of PTBP1 protein, and the Saos-2 cell line with the highest PTBP1 relative expression level was selected for subsequent experiments. Saos-2 cells were divided into 5 groups: si-NC group [transfected with negative control siRNA (NC siRNA)], si-PTBP1 group (transfected with PTBP1 siRNA), solvent control+si-NC group [treated with dimethyl sulfoxide(DMSO) to a final concentration of 0.1% and transfected with NC siRNA], solvent control+si-PTBP1 group (treated with 0.1% DMSO and transfected with PTBP1 siRNA), and SF1670+si-PTBP1 group (treated with 10 μmol/L SF1670, a PTEN inhibitor, and transfected with PTBP1 siRNA). CCK-8 assay was used to detect the cell proliferation [expressed as the absorbance value at 490 nm ( A490)]; TUNEL staining was used to detect apoptosis ability, and apoptosis rate was calculated; Western blotting was used to detect the expression levels of relevant proteins. Results:The relative expression level of PTBP1 protein in osteosarcoma MG63, U2-OS, Saos-2, 143B, and osteoblast cell line hFOB1.19 cells was 0.80±0.12, 0.55±0.08, 1.04±0.10, 0.49±0.07, and 0.26±0.05, respectively, and the difference was statistically significant ( F = 47.10, P < 0.001). CCK-8 assay result showed that the A490 value of Saos-2 cells in si-NC group and si-PTBP1 group was 1.00±0.05 and 0.55±0.07, respectively, and the difference was statistically significant ( t = 10.46, P < 0.001). The result of TUNEL staining showed that the apoptosis rate was (1.26±0.05)% and (12.86±0.87)%, respectively in si-NC group and si-PTBP1 group, and the difference was statistically significant ( t = 26.62, P < 0.001). Western blotting showed that PTEN relative expression level of Saos-2 cell in si-PTBP1 group was higher than that in si-NC group (1.02±0.05 vs. 0.26±0.04, t = 23.74, P < 0.001), and the relative expression level of p-Akt in si-PTBP1 group was lower than that in si-NC group (0.22±0.03 vs. 1.00±0.07, t = 20.48, P < 0.001). In the solvent control+si-NC, solvent control+si-PTBP1, and SF1670+si-PTBP1 groups, the relative expression level of p-Akt protein was 0.96±0.07, 0.21±0.02, and 0.79±0.03, respectively, and the difference was statistically significant ( F = 299.30, P < 0.001), and the relative expression level of PTEN protein was 0.26±0.02, 0.98±0.08, and 0.96±0.11, respectively, and the difference was statistically significant ( F = 106.80, P < 0.001). The A490 values were 1.00±0.10, 0.53±0.08, and 0.89±0.08, respectively in the solvent control+si-NC, solvent control+si-PTBP1, and SF1670+si-PTBP1 groups, and the difference was statistically significant ( F = 31.81, P < 0.001); apoptosis rates were (1.27±0.11)%, (13.26±0.89)%, and (4.86±0.37)%, respectively, and the difference was statistically significant ( F = 482.90, P < 0.001). Conclusions:Knockdown of PTBP1 inhibits the proliferation and promotes the apoptosis of osteosarcoma cells, and this effect may be associated with the regulation of the PTEN-Akt pathway.
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