anti pdk1  (Cell Signaling Technology Inc)

Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: PDK1 overexpression impaired the anti-proliferation effect of gramine on OC cells. (A) Western blotting assay determining the effect of gramine on PDK1 expressions. SK-OV-3 and OV-90 cells were incubated with gramine (50, 100 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the untreated control. (B) Western blotting assay assessing the transfection efficiency of PDK1 overexpression lentivirus. *** P<0.001 vs. the EV control. (C) Cell counting kit-8 assay detecting the effect of PDK1 overexpression on cell viability. (D) Clone formation assay assessing the effect of PDK1 overexpression on colony formation ability. (E) Western blotting assay examining the effect of PDK1 overexpression on Ki67 expression. * P<0.05, ** P<0.01 vs. the EV control or gramine-treated EV group (C-E). Data represent the mean ± SD of three independent experiments. EV, empty vector; OV, overexpression.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: Over Expression, Western Blot, Incubation, Control, Transfection, Cell Counting, Tube Formation Assay, Colony Assay, Expressing, Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: PDK1 overexpression impaired the pro-apoptotic effect of gramine on ovarian cancer cells. (A) TUNEL assay evaluating the effect of PDK1 overexpression on cell apoptosis. Scale bar, 200 µm. (B) Western blotting assay determining the effect of PDK1 overexpression on Bax and Bcl2 expressions. PDK1 overexpressing SK-OV-3 and OV-90 cells were treated with gramine (100 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the EV control or gramine-treated EV group. Data represent the mean ± SD of three independent experiments. EV, empty vector; OV, overexpression.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: Over Expression, TUNEL Assay, Western Blot, Control, Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: PDK1/AKT signaling pathway was involved in the anti-migratory and anti-invasive effects of gramine in ovarian cancer cells. (A) Wound healing and (B) transwell invasion assays detecting the effect of PDK1 overexpression on the abilities of cell migration and invasion respectively. Scale bar, 200 µm. (C) Western blotting assay determining the effect of PDK1 overexpression on N-cadherin, vimentin and p-AKT (Thr308) expressions. PDK1 overexpressing SK-OV-3 and OV-90 cells were incubated with gramine (50 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the EV control or gramine-treated EV group. Data represent the mean ± SD of three independent experiments. EV, empty vector; OV, overexpression.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: Over Expression, Migration, Western Blot, Incubation, Control, Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: Gramine downregulates PDK1/AKT linked to cisplatin sensitization of OC cells. (A) CCK-8 assay detecting the effect of cisplatin on cell viability. SK-OV-3 and SK-OV-3-R cells were incubated with cisplatin (5, 10, 20, 40, 80 µM) for 24 h. * P<0.05, ** P<0.01 vs. the untreated control. (B) CCK-8 assay detecting the effect of gramine and cisplatin on cell viability. SK-OV-3-R cells were incubated with gramine (50 µM) and cisplatin (5, 10, 20, 40, 80 µM) for 24 h. * P<0.05 vs. the DMSO group at the identical cisplatin concentration. (C) Clone formation assay assessing the effect of gramine and cisplatin on colony formation ability. (D) TUNEL assay evaluating the effect of gramine and cisplatin on cell apoptosis. Scale bar, 200 µm. (E) Wound healing and transwell invasion assays detecting the effect of gramine and cisplatin on the abilities of cell migration and invasion respectively. Scale bar, 200 µm. (F) Western blotting assay determining the effect of gramine and cisplatin on the expressions of PDK1 and p-AKT (Thr308). SK-OV-3-R cells were incubated with gramine (50 µM) and cisplatin (80 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the cisplatin group (C-F). Data represent the mean ± SD of three independent experiments. CCK-8, cell counting kit-8.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: CCK-8 Assay, Incubation, Control, Concentration Assay, Tube Formation Assay, Colony Assay, TUNEL Assay, Migration, Western Blot, Cell Counting