Journal: American Journal of Cancer Research

Article Title: Peruvoside is a novel Src inhibitor that suppresses NSCLC cell growth and motility by downregulating multiple Src-EGFR-related pathways

doi:

Figure Lengend Snippet: Effects of peruvoside on Src, EGFR and STAT3 expression and the viability of different cell lines. A. Chemical structure of peruvoside. B. Inhibition of A549 cell viability by peruvoside at different concentrations and time points as determined by PrestoBlue staining. The results are presented as percentages of the vehicle control (0 µM, 0.1% DMSO). The IC50 values of the designated time points are shown in Table S1. Each experiment was performed independently and in triplicate. *P < 0.05 compared with the vehicle control. C. Inhibition of the protein expression and phosphorylation of Src, EGFR, and STAT3 by peruvoside in A549 cells. Left panel: cells were treated with the indicated concentrations of peruvoside for 24 h and then subjected to western blotting. Right panel: cells were treated with 50 nM peruvoside for the specified time and then subjected to western blotting. D. Inhibition of Src, EGFR, and STAT3 expression and phosphorylation by peruvoside in H3255 (left panel) and H1975 (right panel) cells. The cells were treated with the indicated concentrations of peruvoside for 24 h and then analysed by western blotting. The protein expression levels were quantified by ImageJ software (NIH) and are directly shown below the gel image. Ctrl: 0.1% DMSO; Da100: 100 nM dasatinib as a positive control. GAPDH served as a loading control.

Article Snippet: The human bronchial epithelial cell line BEAS2B (ATCC CRL-9609) and human lung adenocarcinoma cell lines A549 (ATCC CCL-185; EGFR wild-type, TKI resistant) and H1975 (ATCC CRL-5908; EGFR L858R+T790M, TKI resistant) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Inhibition, Staining, Control, Phospho-proteomics, Western Blot, Software, Positive Control

Journal: American Journal of Cancer Research

Article Title: Peruvoside is a novel Src inhibitor that suppresses NSCLC cell growth and motility by downregulating multiple Src-EGFR-related pathways

doi:

Figure Lengend Snippet: Comparison of the efficacies of peruvoside and dasatinib in NSCLC cell lines with EGFR wild-type and EGFR mutation statuses. The lung cancer cell lines A549 (A), PC9 (B), PC9/gef (C), H3255 (D) and H1975 (E) were treated with the same concentration (100 nM) of peruvoside and dasatinib to compare their inhibitory effects on proliferation at the indicated time points, as determined by the PrestoBlue cell viability assay. The results are presented as percentages of the vehicle control (0 µM, 0.1% DMSO). Each experiment was performed independently and in triplicate. *P < 0.05 and ** P<0.0001 compared with the vehicle control.

Article Snippet: The human bronchial epithelial cell line BEAS2B (ATCC CRL-9609) and human lung adenocarcinoma cell lines A549 (ATCC CCL-185; EGFR wild-type, TKI resistant) and H1975 (ATCC CRL-5908; EGFR L858R+T790M, TKI resistant) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Comparison, Mutagenesis, Concentration Assay, Viability Assay, Control

Journal: American Journal of Cancer Research

Article Title: Peruvoside is a novel Src inhibitor that suppresses NSCLC cell growth and motility by downregulating multiple Src-EGFR-related pathways

doi:

Figure Lengend Snippet: Suppressive effects of peruvoside on anchorage-dependent and anchorage-independent growth and motility. The colony formation assay was used to determine the inhibitory effect of peruvoside on clonogenicity. A. Anchorage-dependent cell growth of the A549 cell line. Colonies with diameters ≥ 1 mm were counted. B. Anchorage-independent cell growth of the A549 cell line. Colonies with diameters ≥1 mm were counted. Each experiment was performed independently and in triplicate. 0 nM: 0.1% DMSO. C. Inhibitory effect of peruvoside on cancer cell migration, as determined by the Transwell assay. D. Inhibitory effect of peruvoside on cancer cell invasion, as measured by the Transwell assay. Each experiment was performed independently and in triplicate. *P < 0.05 compared with the vehicle control.

Article Snippet: The human bronchial epithelial cell line BEAS2B (ATCC CRL-9609) and human lung adenocarcinoma cell lines A549 (ATCC CCL-185; EGFR wild-type, TKI resistant) and H1975 (ATCC CRL-5908; EGFR L858R+T790M, TKI resistant) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Colony Assay, Migration, Transwell Assay, Control

Journal: American Journal of Cancer Research

Article Title: Peruvoside is a novel Src inhibitor that suppresses NSCLC cell growth and motility by downregulating multiple Src-EGFR-related pathways

doi:

Figure Lengend Snippet: Effects of peruvoside on tumour growth and drug synergism. The indicated number of live A549 cells was subcutaneously injected into mice divided into vehicle-treated (n = 6) and drug-treated groups (n = 6). A. The tumour volumes were measured every 7 days. B. Peruvoside decreased the tumour weight. The data are presented as the means ± standard deviations. C. The p-Src level and distribution in murine tumour tissues were determined by immunohistochemical staining and observed under a light microscope (400 × magnification). The control represents 0.1% DMSO, and peruvoside was administered at 0.1 mg/kg. The scale bars represent 20 μm. *P < 0.05 compared with the vehicle control (0.1% DMSO). D. Synergistic effects of peruvoside and gefitinib on lung cancer cell lines, as determined by the cell viability assay. The indicated combinations of peruvoside and gefitinib were used to treat the gefitinib-resistant lung adenocarcinoma cell lines A549, PC9/gef and H1975 for 72 h. The CI was calculated using CalcuSyn software. Each experiment was performed independently and was repeated three times.

Article Snippet: The human bronchial epithelial cell line BEAS2B (ATCC CRL-9609) and human lung adenocarcinoma cell lines A549 (ATCC CCL-185; EGFR wild-type, TKI resistant) and H1975 (ATCC CRL-5908; EGFR L858R+T790M, TKI resistant) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Injection, Immunohistochemical staining, Staining, Light Microscopy, Control, Viability Assay, Software

Journal: American Journal of Cancer Research

Article Title: Peruvoside is a novel Src inhibitor that suppresses NSCLC cell growth and motility by downregulating multiple Src-EGFR-related pathways

doi:

Figure Lengend Snippet: Effects of peruvoside on the expression and phosphorylation of Src and related proteins. A549 (A), H3255 (B), and H1975 (C) lung cancer cells were treated with peruvoside at the indicated concentrations for 24 h. The expression and phosphorylation levels of PI3K, AKT, MEK, ERK, FAK, JNK, Paxillin, and p130cas were measured by immunoblot analysis using the corresponding antibodies. The protein expression levels were quantified by ImageJ software (NIH) and are directly shown below the gel image. GAPDH was used as an internal control. Ctrl: 0.1% DMSO. Each experiment was performed independently and in triplicate.

Article Snippet: The human bronchial epithelial cell line BEAS2B (ATCC CRL-9609) and human lung adenocarcinoma cell lines A549 (ATCC CCL-185; EGFR wild-type, TKI resistant) and H1975 (ATCC CRL-5908; EGFR L858R+T790M, TKI resistant) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Phospho-proteomics, Western Blot, Software, Control

Journal: American Journal of Cancer Research

Article Title: Peruvoside is a novel Src inhibitor that suppresses NSCLC cell growth and motility by downregulating multiple Src-EGFR-related pathways

doi:

Figure Lengend Snippet: Peruvoside inhibits Src and Src-related gene transcription. A. Repressive effects of peruvoside on Src, EGFR, STAT3, and FAK transcription in A549 cells, as determined by real-time RT-PCR. The relative gene expression levels were calculated using the comparative CT method (2-ΔΔCT). TBP: internal control. Each experiment was performed independently and in triplicate. *P < 0.05 compared with the control (0 nM: 0.1% DMSO). B. Role of the ubiquitin-proteasome system in peruvoside-mediated decreases in protein expression. MG132 and/or peruvoside was administered to A549 cells for 24 h before the cells were subjected to western blotting. GAPDH served as a loading control. Protein expression was quantified with ImageJ software (NIH), and the results are shown directly below the gel graph. C. A hypothetical model illustrating the role of peruvoside in suppressing lung cancer tumorigenesis. Peruvoside significantly suppressed the phosphorylation of Src, EGFR, and STAT3 in different NSCLC cell lines (A549, H3255 and H1975 cells) regardless of their EGFR mutation status and had a synergistic effect when combined with gefitinib. Furthermore, the inhibitory effects of peruvoside on lung cancer progression might be attributed to its ability to regulate multiple proteins, including Src, PI3K, JNK, Paxillin, p130cas, and EGFR. By combining these effects, peruvoside can effectively inhibit tumorigenesis.

Article Snippet: The human bronchial epithelial cell line BEAS2B (ATCC CRL-9609) and human lung adenocarcinoma cell lines A549 (ATCC CCL-185; EGFR wild-type, TKI resistant) and H1975 (ATCC CRL-5908; EGFR L858R+T790M, TKI resistant) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Gene Expression, Control, Ubiquitin Proteomics, Expressing, Western Blot, Software, Phospho-proteomics, Mutagenesis

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Lung cancer cell viability is suppressed by Sch A in a concentration- and time-dependent manner. Sch A decreased (A) A549 and (B) HCC827 cell viability in a concentration-dependent manner, according to Cell Counting Kit-8 analysis. Sch A significantly reduced the viability of (C) A549 and (D) HCC827 cells at 24, 48 and 72 h. ***P<0.001 vs. Con. Con, control; Sch A, Schisantherin A; OD, optical density.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Concentration Assay, Cell Counting, Control

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Sch A induces cell death and cell cycle arrest in A549 and HCC827 cells. Sch A considerably increased the death of (A) A549 and (B) HCC827 cells in comparison with the control group, according to flow cytometric analysis. Compared with Con, (C) A549 and (D) HCC827 cells presented a significant decrease in the number of G 2 + S-phase cells and a significant increase in the number of G 1 -phase cells. Fer-1 and DFO were able to significantly reverse the Sch A-induced decrease in (E) A549 and (F) HCC827 cell viability, according to the results of the Cell Counting Kit-8 analysis. *P<0.05, **P<0.01 and ***P<0.001 vs. Con; ## P<0.01 and ### P<0.001 vs. Sch A. DFO, deferoxamine; Con, control; Sch A, Schisantherin A; Fer-1, ferrostatin-1; Nec-1, necrostatin-1; 3-MA, 3-methyladenine.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Comparison, Control, Cell Counting

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: In A549 and HCC827 cells, ferroptosis inhibitors prevent Sch A-induced apoptosis. Fer-1 and DFO were shown by JC-1 staining to reverse the Sch A-induced MMP reduction in (A) A549 and (B) HCC827 cells (scale bar, 30 µm). Fer-1 and DFO inhibited the Sch A-induced increase in MDA in (C) A549 and (D) HCC827 cells. Fer-1 and DFO counteracted the Sch A-induced reduction in GSH in (E) A549 and (F) HCC827 cells. Reverse transcription-quantitative PCR analysis revealed that Fer-1 and DFO reversed the Sch A-induced increase in Chac1 and ptgs2 levels in (G) A549 and (H) HCC827 cells. *P<0.05, **P<0.01 and ***P<0.001 vs. Con; # P<0.05, ## P<0.01 and ### P<0.001 vs. Sch A. Con, control; Chac1, glutathione-specific γ-glutamylcyclotransferase 1; ptgs2, prostaglandin-endoperoxide synthase 2; Fer-1, ferrostatin-1; DFO, deferoxamine; Sch A, Schisantherin A; MDA, malondialdehyde; GSH, glutathione.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Sch A induces ferroptosis in lung cancer cells by activating YAP signaling. (A) GeneCards intersection analysis revealed 739 overlapping genes associated with both ferroptosis and NSCLC. (B) Transcription factors among the intersecting genes were analyzed to identify key node genes. Reverse-transcription-quantitative PCR analysis revealed that Sch A increased the mRNA expression levels of YAP1 in (C) A549 and (D) HCC827 cells. Western blot analysis revealed that Sch A increased the expression levels of TfR, ACSL4 and YAP in (E) A549 and (F) HCC827 cells. *P<0.05 and **P<0.01 vs. Con. Con, control; YAP, yes-associated protein; Sch A, Schisantherin A; TfR, transferrin receptor; ACSL4, acyl-CoA synthase long-chain family member 4; NSCLC, non-small cell lung cancer.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Silencing of YAP reverses Sch A-induced ferroptosis. Western blot analysis showed that the expression of YAP was decreased in (A) A549 and (B) HCC827 cells. In (C) A549 and (D) HCC827 cells, si-YAP decreased the expression of YAP and downstream signaling molecules. Fe 2+ accumulation was induced by Sch A in (E) A549 and (F) HCC827 cells, and the increase in Fe 2+ was reversed by YAP silencing, as demonstrated by FerroOrange staining (scale bar, 10 µm). After silencing YAP, the Sch A-induced increase in Fe 2+ and MDA in (G) A549 and (H) HCC827 cells was reduced. *P<0.05, **P<0.01 and ***P<0.001 vs. NC; ### P<0.001 vs. Sch A. NC, negative control; YAP, yes-associated protein; Sch A, Schisantherin A; si, small interfering RNA; MDA, malondialdehyde; ACSL4, acyl-CoA synthase long-chain family member 4; TfR, transferrin receptor.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Western Blot, Expressing, Staining, Negative Control, Small Interfering RNA