Journal: Translational Cancer Research

Article Title: Obovatol induces apoptosis in breast cancer by downregulating the PI3K/Akt pathway

doi: 10.21037/tcr-2025-1-2627

Figure Lengend Snippet: Ob diminished the activity of the PI3K/Akt pathway in BC cells. (A) Volcano plot showing DEGs in MDA-MB-231 cells between the Ob group and control groups. (B) Gene Ontology enrichment analysis revealed that the DEGs were strongly enriched in specific biological processes, cellular components, and molecular functions in the Ob-treated BC cells. (C) KEGG enrichment analysis revealed altered signaling pathways in the Ob-treated BC cells. (D) A clustering heatmap analysis was conducted to identify the DEGs associated with the PI3K/Akt pathway. (E) The relative protein expression of p-PI3K, PI3K, p-Akt, and Akt was assessed by WB assay. (F) The relative protein expression of p-PI3K, PI3K, p-Akt, and Akt was assessed by WB assay after the treatment with 50 µM of Ob and 20 µM of 740Y-P for 48 h. *, P<0.05; **, P<0.01 ( vs. control group); # , P<0.05; ## , P<0.01 ( vs. Ob group). ADR, doxorubicin hydrochloride; Akt, protein kinase B; BC, breast cancer; Con, control; DEGs, differentially expressed genes; KEGG, Kyoto Encyclopedia of Genes and Genomes; Ob, obovatol; p-Akt, phosphorylated Akt; p-PI3K, phosphorylated PI3K; PI3K, phosphoinositide 3-kinase; WB, Western blotting.

Article Snippet: Phosphoinositide 3-kinase (PI3K; bs-10657R), phosphorylated PI3K (p-PI3K; bs-6417R), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; bs-33033M), mouse secondary antibody (bs-0296G-HRP), and rabbit secondary antibody (bs-0295G-HRP) were supplied by Bioss (Beijing, China). β-tubulin (A12289) was acquired from ABclonal (Wuhan, China). β-tubulin and GAPDH were used as loading controls.

Techniques: Activity Assay, Control, Protein-Protein interactions, Expressing, Western Blot

Journal: Translational Cancer Research

Article Title: Obovatol induces apoptosis in breast cancer by downregulating the PI3K/Akt pathway

doi: 10.21037/tcr-2025-1-2627

Figure Lengend Snippet: Ob suppressed invasion and induced the apoptosis of BC cells by inhibiting the PI3K/Akt pathway. (A) OD values at 450 nm for BC cells were measured by CCK-8 assay following treatment with Ob and 740Y-P for durations of 0, 24, 48, 72, and 96 h. (B) Inhibition of invasion ability of BC cells was assessed by Transwell assay following treatment with Ob and 740Y-P (crystal violet staining). (C) EMT-relative mRNA expression of E-cadherin, N-cadherin, and vimentin was assessed by RT-qPCR assay following treatment with Ob and 740Y-P. (D) Apoptosis induced by Ob in BC cells was assessed by flow cytometry. (E) Relative mRNA expression of Bcl-2 and Bax associated with apoptosis was assessed by RT-qPCR assay. *, P<0.05; **, P<0.01; ***, P<0.001 ( vs. control group); # , P<0.05; ## , P<0.01 ( vs. Ob group), n=3. ADR, doxorubicin hydrochloride; Akt, protein kinase B; Annexin V-APC, Annexin V-allophycocyanin conjugate; Bax, Bcl-2-associated X protein; BC, breast cancer; Bcl-2, B-cell lymphoma 2; CCK-8, cell counting kit-8; Con, control; EMT, epithelial-mesenchymal transition; Ob, obovatol; OD, optical density; PI, propidium iodide; PI3K, phosphoinositide 3-kinase; RT-qPCR, real-time quantitative polymerase chain reaction.

Article Snippet: Phosphoinositide 3-kinase (PI3K; bs-10657R), phosphorylated PI3K (p-PI3K; bs-6417R), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; bs-33033M), mouse secondary antibody (bs-0296G-HRP), and rabbit secondary antibody (bs-0295G-HRP) were supplied by Bioss (Beijing, China). β-tubulin (A12289) was acquired from ABclonal (Wuhan, China). β-tubulin and GAPDH were used as loading controls.

Techniques: CCK-8 Assay, Inhibition, Transwell Assay, Staining, Expressing, Quantitative RT-PCR, Flow Cytometry, Control, Cell Counting, Real-time Polymerase Chain Reaction