Journal: Oncology Letters

Article Title: Sakurasosaponin inhibits lung cancer cell proliferation by inducing autophagy via AMPK activation

doi: 10.3892/ol.2023.14088

Figure Lengend Snippet: S-saponin inhibits non-small cell lung cancer cell proliferation. (A and B) A549 and H1299 cells were treated with various concentrations of S-saponin for 24 or 48 h, and cell proliferation was assessed using a Cell Counting Kit-8 cell viability assay. Data represent the mean ± SD of three independent experiments. (C and D) The clonogenic assay showing the effects of S-saponin on colony formation in A549 and H1299 cells. After treatment with S-saponin in a dose-dependent manner, the media was changed after 24 h. The media was then changed once every 2–3 days; the cells were fixed after 7 days of observation and counted following the staining with crystal violet. Representative images of colonies are shown, and quantitative data represent the mean ± SD of three independent experiments. *P<0.05, **P<0.01 and ***P<0.001 vs. the control group. S-saponin, sakurasosaponin.

Article Snippet: The human NSCLC cell lines, A549 (cat. no. CCL-185) and H1299 (cat. no. CRL-5803) were obtained from the American Type Culture Collection.

Techniques: Cell Counting, Viability Assay, Clonogenic Assay, Staining, Control

Journal: Oncology Letters

Article Title: Sakurasosaponin inhibits lung cancer cell proliferation by inducing autophagy via AMPK activation

doi: 10.3892/ol.2023.14088

Figure Lengend Snippet: S-saponin does not induce apoptosis in A549 and H1299 cells. A549 and H1299 cells were treated with S-saponin (A) in a dose-dependent manner for 24 h and (B) in a time-dependent manner (5 µg/ml). The cells were subjected to western blotting using the PARP and tubulin antibodies. The experiment was independently performed thrice, quantified through the ImageJ software and normalized with tubulin. (C) A549 and H1299 cells were treated with S-saponin (5 µg/ml) for 24 h. The cell viability of S-saponin-treated A549 and H1299 cells was determined through FACS analysis using annexin V/PI staining. S-saponin, sakurasosaponin; PARP, poly (adenosine diphosphate-ribose) polymerase.

Article Snippet: The human NSCLC cell lines, A549 (cat. no. CCL-185) and H1299 (cat. no. CRL-5803) were obtained from the American Type Culture Collection.

Techniques: Western Blot, Software, Staining

Journal: Oncology Letters

Article Title: Sakurasosaponin inhibits lung cancer cell proliferation by inducing autophagy via AMPK activation

doi: 10.3892/ol.2023.14088

Figure Lengend Snippet: S-saponin induces autophagy in non-small cell lung cancer cells. A549 and H1299 cells were treated with S-saponin (A) in a dose-dependent manner for 24 h and (B) in a time-dependent manner (5 µg/ml). The cells were subjected to western blotting using the indicated antibodies. The experiment was independently performed thrice, quantified through the ImageJ software and normalized with full LC3 band. (C) A549 and H1299 cells were transfected with GFP-LC3 vector and treated with or without S-saponin for 12 h. After 12 h of culture, the cells were counterstained with Hoechst 33342. Fluorescence microscopy images of GFP-LC3 puncta formation were recorded using a Leica Confocal laser scanning microscope. Scale bar, 10 µm; magnification, ×40. S-saponin, sakurasosaponin.

Article Snippet: The human NSCLC cell lines, A549 (cat. no. CCL-185) and H1299 (cat. no. CRL-5803) were obtained from the American Type Culture Collection.

Techniques: Western Blot, Software, Transfection, Plasmid Preparation, Fluorescence, Microscopy, Laser-Scanning Microscopy

Journal: Oncology Letters

Article Title: Sakurasosaponin inhibits lung cancer cell proliferation by inducing autophagy via AMPK activation

doi: 10.3892/ol.2023.14088

Figure Lengend Snippet: S-saponin activates the AMPK signaling pathway in A549 and H1299 non-small cell lung cancer cells. A549 and H1299 cells were treated with S-saponin (A) in a dose-dependent manner for 24 h and (B) in a time-dependent manner (5 µg/ml). The cells were subjected to western blotting using the phosphor-AMPKα and AMPKα antibodies. Tubulin was used as the loading control. The experiment was independently performed thrice, quantified through the ImageJ software and normalized with AMPK. *P<0.05 and **P<0.01 vs. 0 µΜ or 0 h. S-saponin, sakurasosaponin; AMPK, adenosine monophosphate-activated protein kinase.

Article Snippet: The human NSCLC cell lines, A549 (cat. no. CCL-185) and H1299 (cat. no. CRL-5803) were obtained from the American Type Culture Collection.

Techniques: Western Blot, Control, Software

Journal: Oncology Letters

Article Title: Sakurasosaponin inhibits lung cancer cell proliferation by inducing autophagy via AMPK activation

doi: 10.3892/ol.2023.14088

Figure Lengend Snippet: Pharmacologic inhibition of the AMPK pathway attenuates S-saponin-induced autophagy and cell proliferation inhibition. A549 and H1299 cells were pretreated with 20 µM Compound C for 30 min, followed by 5 µg/ml S-saponin for 24 h. (A) The cells were subjected to western blotting using p-AMPKα, AMPKα, and LC3 II antibodies. Tubulin was used as the loading control. The experiment was independently performed thrice, quantified through the ImageJ software and normalized with AMPK or full LC3 band. (B) A549 and H1299 cells were pretreated with 20 µM Compound C for 30 min, followed by 5 µg/ml S-saponin for 24 h. Cell proliferation was assessed using a Cell Counting Kit-8 cell viability assay. Data represent the mean ± SD of three independent experiments. *P<0.05 and **P<0.001 compared with the control group. AMPK, adenosine monophosphate-activated protein kinase; S-saponin, sakurasosaponin; p-, phosphorylated.

Article Snippet: The human NSCLC cell lines, A549 (cat. no. CCL-185) and H1299 (cat. no. CRL-5803) were obtained from the American Type Culture Collection.

Techniques: Inhibition, Western Blot, Control, Software, Cell Counting, Viability Assay

Journal: Oncology Letters

Article Title: Sakurasosaponin inhibits lung cancer cell proliferation by inducing autophagy via AMPK activation

doi: 10.3892/ol.2023.14088

Figure Lengend Snippet: Knockdown of AMPK attenuates S-saponin-induced autophagy and cell proliferation inhibition. (A) A549 and H1299 cells were transfected with control siRNA or siRNA-AMPKα and treated with or without 5 µg/ml S-saponin. The cells were subjected to western blotting using p-AMPKα, AMPKα, and LC3 II antibodies. Tubulin was used as the loading control. The experiment was independently performed thrice, quantified through the ImageJ software and normalized with AMPK or full LC3 band. (B) A549 and H1299 cells were transfected with control siRNA or siRNA-AMPK and treated with or without 5 µg/ml S-saponin. Following the treatment with S-saponin, the media was changed once after 24 h, and then once every 2 days; the cells were fixed after 4 days of observation and counted following the staining with crystal violet. Representative images of colonies are shown, and quantitative data represent the mean ± SD of three independent experiments. *P<0.05 and ***P<0.001 compared with control group. AMPK, adenosine monophosphate-activated protein kinase; S-saponin, sakurasosaponin; si, small interfering; p-, phosphorylated.

Article Snippet: The human NSCLC cell lines, A549 (cat. no. CCL-185) and H1299 (cat. no. CRL-5803) were obtained from the American Type Culture Collection.

Techniques: Knockdown, Inhibition, Transfection, Control, Western Blot, Software, Staining

Journal: Molecular cancer therapeutics

Article Title: GnRH-R targeted lytic peptide sensitizes BRCA wild-type ovarian cancer to PARP inhibition

doi: 10.1158/1535-7163.MCT-18-0770

Figure Lengend Snippet: (A) A network of downregulated/upregulated proteins in combination group compared to olaparib group was determined using Netwalker analysis. (B) Expression of PI3K/AKT, p-PI3K/p-AKT, PARP, Cleaved PARP, BRCA1, BRCA2, and RAD 51 after treatment. (C) Expression and localization of GnRH-R in BRCA1 mutant (BRCA1m) COV362 and MDA-MB-436 cells and BRCA2 mutant (BRCA2m) KURAMOCHI cells. The fixed cells without permeabilization (left panels) were visualized using confocal microscopy (TCS SP5 MP; Leica Microsystems, Buffalo Grove, IL). The fixed and permeabilized cells (right panels) were visualized using a laser-scanning microscope (Leica). (Scale bar: 50 μM). (D) cell viability of COV362, MDA-MB-436, and KURAMOCHI cells after treatment with EP-100, olaparib, or EP-100 plus olaparib for 72 h. (E) CI values are shown as in Fa-CI plots.

Article Snippet: The primary antibody dilution factors were as follows according to manufacturer’s instructions: anti-GnRH-R (1:1,000, Abcam, Cambridge, UK, Cat: ab183079), RAD 51 (1:10,000, Abcam, Cambridge, UK, Cat: ab133534), phosphorylated PI3K p85 (1:1,000, Cat: 4228s), PI3K (1:1,000, Cat: 4292s), phosphorylated AKT Ser473 (1:1,000, Cat: 9271s), AKT (1:1,000, Cat: 9272s), PARP (1:1,000, Cat: 9532s), BRCA1 (1:1,000, Cat: 9025s) and BRCA2 (1:1,000, Cat: 10741s) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Mutagenesis, Confocal Microscopy, Laser-Scanning Microscopy