Journal: Redox Report : Communications in Free Radical Research

Article Title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

doi: 10.1080/13510002.2025.2588086

Figure Lengend Snippet: BS affects NF-κB, p38/JNK, and PI3K-Akt signaling pathways in lung cancer. (a–d) A549 and H1299 cells were treated with BS (0, 5, 10 μM) for 24 h, followed by western blot. Error bars are means ± std. * P < 0.05, ** P < 0.01, *** P < 0.001, compared with the control group by One-way ANOVA (n = 3).

Article Snippet: The primary antibodies used for immunoblotting analysis are as follows: TrxR1 (Proteintech, 11117-1-AP), Trx1 (Proteintech, 14999-1-AP), HBP1 (Proteintech, 11746-1-AP), DNMT1 (Proteintech, 24206-1-AP), Bcl-2 (Proteintech, 12789-1-AP), Bax (Proteintech, 50599-2-Ig), β-actin (Bioss, bs-0061R), Flag (Sigma-Aldrich, F1804), HA (Covance, MMS-101P), p53 (Santa, sc-126), p21 (MBL, K0081-3), p27 (MBL, K0082), γ-H2AX (CST, 9718), NF-κB (Abcam, ab32536), p-NF-κB(CST, 3033), p38 (Santa, sc-7972), p-p38 (Santa, sc-101759), JNK (MCE, HY- P80728 ), p-JNK (Immunoway, YP0157), Akt (Santa, sc-5298), phospho-Akt (CST, 4060).

Techniques: Protein-Protein interactions, Western Blot, Control

Journal: Redox Report : Communications in Free Radical Research

Article Title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

doi: 10.1080/13510002.2025.2588086

Figure Lengend Snippet: Schematic model of lung cancer inhibition by BS. The TrxR/Trx inhibitor butaselen (BS) can inhibit lung cancer by inducing ROS-dependent apoptosis. The inactivation of the NF-κB and PI3K-Akt signaling pathways, along with the activation of the p38/JNK signaling pathway, contributes to the anti-cancer effects of BS on lung cancer. Although p53 itself is not activated by BS, the HBP1/DNMT1/p21/γ-H2AX/Bcl-2/Bax signaling pathway is activated by BS and contributes to its tumor-inhibitory role. Further mechanistic studies revealed HBP1 as a novel target of the Trx system. The Trx system inversely associates with HBP1 in lung cancer and regulates HBP1 expression at the post-translational level. Under normal conditions, TrxR1 catalyzes the reduction of Trx1 by utilizing NADPH. In its reduced form, Trx1 interacts with HBP1, promoting the ubiquitination of HBP1, which leads to its proteasomal degradation and maintains a low level of HBP1 within cancer cells. Treatment with butaselen inhibits the activity of TrxR1 in lung cancer cells, resulting in the oxidation of Trx1 and the subsequent excessive generation of ROS. HBP1 is activated after being released by the oxidized Trx1 and escaping proteasomal degradation. The activated HBP1 inhibits the expression of DNMT1 and elevates Bax. The decreased DNMT1 further results in the demethylation of the whole genome as well as the promoters of p21 and HOXA9. Ultimately, the upregulation of p21 and γ-H2AX, along with the downregulation of DNMT1 and Bcl-2/Bax, contributes to the apoptosis of lung cancer cells induced by BS. Taken together, the TrxR/Trx inhibitor butaselen inhibits lung cancer by promoting ROS-induced apoptosis through the NF-κB, PI3K-Akt, p38/JNK, and HBP1/DNMT1 signaling pathways.

Article Snippet: The primary antibodies used for immunoblotting analysis are as follows: TrxR1 (Proteintech, 11117-1-AP), Trx1 (Proteintech, 14999-1-AP), HBP1 (Proteintech, 11746-1-AP), DNMT1 (Proteintech, 24206-1-AP), Bcl-2 (Proteintech, 12789-1-AP), Bax (Proteintech, 50599-2-Ig), β-actin (Bioss, bs-0061R), Flag (Sigma-Aldrich, F1804), HA (Covance, MMS-101P), p53 (Santa, sc-126), p21 (MBL, K0081-3), p27 (MBL, K0082), γ-H2AX (CST, 9718), NF-κB (Abcam, ab32536), p-NF-κB(CST, 3033), p38 (Santa, sc-7972), p-p38 (Santa, sc-101759), JNK (MCE, HY- P80728 ), p-JNK (Immunoway, YP0157), Akt (Santa, sc-5298), phospho-Akt (CST, 4060).

Techniques: Inhibition, Protein-Protein interactions, Activation Assay, Expressing, Ubiquitin Proteomics, Activity Assay

Journal: Biomedicines

Article Title: Extracellular Vesicles from Plasma of Patients with Glioblastoma Promote Invasion of Glioblastoma Cells Even After Tumor Resection

doi: 10.3390/biomedicines12122834

Figure Lengend Snippet: Analysis of influence of EVs on the MAP kinases phosphorylation in astrocytes ( a , b ), GBM 011 cells ( c , d ), and U251 MG cells ( e , f ). The following phosphorylation sites were assayed: pAKT (S473), pERK1/2 (T202/204; T185/Y187), pJNK1/2/3 (T183/Y185), and pp38 (T180/Y182). ( a , c , e ) Cut representative membranes with the bands (marked by frames); ( b , d , f ) quantification of the band intensity reflecting expression of different MAP kinases in the phosphorylated form. Please note that MAP kinases were assayed on the same (pJNK and pp38) or cut (pAKT and pERK) membranes, so they ‘share’ GAPDH. Whole Western blotting membranes are shown in . The quantification data are presented as band intensity (relative to GAPDH) normalized to that of the untreated cells (100%, dashed line) ± SEM (n = 3–8); $ ( p < 0.05) indicates significant difference from the untreated cells according to one-sample Wilcoxon test, followed by post hoc Holm–Sidak’s test; # ( p < 0.05) and ### ( p < 0.001) indicate significant difference between the data groups according to Kruskal–Wallis test followed by post hoc Dunn’s test.

Article Snippet: After that, the membranes were incubated with the rabbit primary antibodies to pAKT (S473) (1:4000, GB150002, ServiceBio, Wuhan, China), pERK1/2 (T202/204; T185/Y187) (1:1000, GB113492, ServiceBio), pJNK1/2/3 (T183/Y185) (1:1000, GB12018, ServiceBio), and pp38 (T180/Y182) (1:1000, GB1133880, ServiceBio) overnight at 4 °C.

Techniques: Expressing, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Silencing Ras-Related C3 Botulinum Toxin Substrate 1 Inhibits Growth and Migration of Hypopharyngeal Squamous Cell Carcinoma via the P38 Mitogen-Activated Protein Kinase Signaling Pathway

doi: 10.12659/MSM.907468

Figure Lengend Snippet: Silencing Rac1 inhibits the activation of P38 MAPK signal in vitro . The levels of MKK3 and p-MKK3 ( A, B ) and P38 and p-P38 ( C, D ) were detected by Western blot analysis. GAPDH served as the internal reference. All experiments were repeated 3 times and the results are presented as mean ±SD. *** P<0.001 compared with the negative control group.

Article Snippet: The membranes were blocked with 5% skim milk or 1% bovine serum albumin, and then incubated with Rac1 antibody, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (1: 1000; Proteintech, Wuhan, China), Cyclin D1 antibody (1: 1000; BOSTER, Wuhan, China), Cyclin E antibody, Cyclin B antibody (1: 500; Bioss, Beijing, China), metal matrix proteinase (MMP)-2 antibody, MMP-9 antibody, B cell lymphoma-2 (Bcl-2) antibody, Bcl-2-associated X protein (Bax) antibody, phosphorylated-MAP kinase (p-MKK3) antibody (1: 500; Sangon Biotech, Shanghai, China), caspase-3 antibody, caspase-9 antibody, poly ADP-ribose polymerase (PARP) antibody (1: 1000; Cell Signaling Technology, Beverly, MA, USA), p38 mitogen-activated protein kinase (MAPK) antibody, p-p38 MAPK antibody (1: 500; KeyGen, Nanjing, China), or MKK3 antibody (1: 300; BOSTER) overnight at 4°C.

Techniques: Activation Assay, In Vitro, Western Blot, Negative Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Silencing Ras-Related C3 Botulinum Toxin Substrate 1 Inhibits Growth and Migration of Hypopharyngeal Squamous Cell Carcinoma via the P38 Mitogen-Activated Protein Kinase Signaling Pathway

doi: 10.12659/MSM.907468

Figure Lengend Snippet: Rac1 silencing inhibits the activation of P38 MAPK signal in vivo . The levels of MKK3 and p-MKK3 ( A, B ) and P38 and p-P38 ( C, D ) in each group were detected by Western blot analysis with GAPDH as the internal reference. All experiments were repeated 3 times. The results are presented as mean ±SD. N=6. *** P<0.001 compared with the negative control group.

Article Snippet: The membranes were blocked with 5% skim milk or 1% bovine serum albumin, and then incubated with Rac1 antibody, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (1: 1000; Proteintech, Wuhan, China), Cyclin D1 antibody (1: 1000; BOSTER, Wuhan, China), Cyclin E antibody, Cyclin B antibody (1: 500; Bioss, Beijing, China), metal matrix proteinase (MMP)-2 antibody, MMP-9 antibody, B cell lymphoma-2 (Bcl-2) antibody, Bcl-2-associated X protein (Bax) antibody, phosphorylated-MAP kinase (p-MKK3) antibody (1: 500; Sangon Biotech, Shanghai, China), caspase-3 antibody, caspase-9 antibody, poly ADP-ribose polymerase (PARP) antibody (1: 1000; Cell Signaling Technology, Beverly, MA, USA), p38 mitogen-activated protein kinase (MAPK) antibody, p-p38 MAPK antibody (1: 500; KeyGen, Nanjing, China), or MKK3 antibody (1: 300; BOSTER) overnight at 4°C.

Techniques: Activation Assay, In Vivo, Western Blot, Negative Control

Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: (A) The migration in wound-healing assay of A549 cells after treatment with DMSO or E2 (10 nM), PPT (10 nM), DPN (10 nM), E2+Ful (1 μM) and Ful (1 μM) for 24 h (magnification 40×); (B) The effect on wound closure (percentage) in A549 cells ( *p < 0.05 vs. control group, #p < 0.05 vs. E2 group ); (C) Representative images of Transwell assays to assess A549 cell invasion and migration; (D) The effect of transwell migration assays in A549 cells after treating with DMSO or E2 (10 nM), PPT (10 nM), DPN (10 nM), E2+Ful (1μM) and Ful(1μM) for 24 h; The effect of transwell migration assays in H1793 cells after treatment with the same drugs for 72 h; (E) Representative images of Transwell assays for assessment of H1793 cell invasion and migration; (F) The effect of transwell migration assays in H1793 cells after treatment with DMSO or E2 (10 nM), PPT (10 nM), DPN (10 nM), E2+Ful (1 μM) and Ful (1 μM) for 24 h. The effect of transwell migration assays in H1793 cells after treatment with the same drugs for 72 h; (G-H) Western blot analysis of ERβ, MMP-2, MMP-9, p-p38MAPK, p38MAPK, pAKT and tAKT protein levels at 48 h in A549 cells, respectively. GAPDH was used as a control. All data are shown as the mean ± SD. Results represent three independent experiments. Student's t-test was carried out ( *p < 0.05 vs. control group, #p < 0.05 vs. E2 group ).

Article Snippet: The antibodies used during Western blots included rabbit anti-human ERα from Proteintech, rabbit anti-human ERβ (1:500) from Proteintech, mouse anti-human MMP-1(1:400) from Santa Cruz, mouse anti-human MMP-2(1:400) from Santa Cruz, rabbit anti-human MMP-9 (1:500) from Proteintech, rabbit anti-human p38MAPK(1:500)/p-p38MAPK(1:500) and rabbit anti-human tAkt (1:500)/pAkt(1:500) from Bioworld Technology.

Techniques: Migration, Wound Healing Assay, Western Blot

Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: (A) Western blot analysis of protein expression of ERβ, MMP-2, p-p38MAPK, p38MAPK, pAKT and tAKT in NSCLC cell line A549 when treating with DMSO or E2 (10 nM), E2+LY294002 (10 nM), LY294002 (10 nM), E2+SB203580 (10 nM) and SB203590 (10 nM) for 48 h. (B) the detection of optical density. GAPDH was used as a control. All data are shown as the mean ± SD. Results represent three independent experiments. Student's t-test was carried out ( *p < 0.05 vs. control group, #p < 0.05 vs. E2 group ).

Article Snippet: The antibodies used during Western blots included rabbit anti-human ERα from Proteintech, rabbit anti-human ERβ (1:500) from Proteintech, mouse anti-human MMP-1(1:400) from Santa Cruz, mouse anti-human MMP-2(1:400) from Santa Cruz, rabbit anti-human MMP-9 (1:500) from Proteintech, rabbit anti-human p38MAPK(1:500)/p-p38MAPK(1:500) and rabbit anti-human tAkt (1:500)/pAkt(1:500) from Bioworld Technology.

Techniques: Western Blot, Expressing

Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: (A) After bilateral ovariectomy A549 cells (5 × 10 6 /100 μL) suspended in PBS were injected into 4-week-old female BALB/c nude mice via the tail vein, mice were randomly divided into five groups (N = 5/group): E2(0.09 mg/kg), PPT(1.05 mg/kg), DPN(1.89 mg/kg), E2+ Ful(1.46 mg/kg) and blank control. The lungs were removed after 4 weeks subcutaneously under drug treatment. Gross appearance of metastatic lung tumor nodes in different groups is indicated by bright yellow arrows. (B) Representative pathological image of metastatic A549 tumor at magnification 100ificatione pathological image of groups (N = 5/group): The number of metastatic nodules in the lungs (lung nodules number in every mouse of each group), metastatic index in different groups mentioned in ‘‘Materials and methods’’ section for the experimental. (C-E) The mean lung wet weight of each group, The number of metastatic nodules in the lungs (lung nodules number in every mouse of each group), metastatic index in different groups mentioned in ‘‘Materials and methods’’ section for the experimental. (F) Protein expression of ERβ, MMP-2, MMP-9, p-p38MAPK, p38MAPK, pAKT and tAKT in murine lung metastatic nodes was analyzed using Western blot, and (G) the detection of optical density. All data are expressed as the mean ± SD. Student's t-test was carried out ( *p < 0.05 vs. control group, #p < 0.05 vs. E2 group ).

Article Snippet: The antibodies used during Western blots included rabbit anti-human ERα from Proteintech, rabbit anti-human ERβ (1:500) from Proteintech, mouse anti-human MMP-1(1:400) from Santa Cruz, mouse anti-human MMP-2(1:400) from Santa Cruz, rabbit anti-human MMP-9 (1:500) from Proteintech, rabbit anti-human p38MAPK(1:500)/p-p38MAPK(1:500) and rabbit anti-human tAkt (1:500)/pAkt(1:500) from Bioworld Technology.

Techniques: Injection, Expressing, Western Blot

Journal: Poultry Science

Article Title: Zinc alleviates thermal stress-induced damage to the integrity and barrier function of cultured chicken embryonic primary jejunal epithelial cells via the MAPK and PI3K/AKT/mTOR signaling pathways

doi: 10.1016/j.psj.2024.103696

Figure Lengend Snippet: Information about primary antibodies.

Article Snippet: p-p38MAPK , 1:2000 , A1459 , ABclonal , China.

Techniques: