Journal: ACS Omega

Article Title: The Role and Efficacy of JNK Inhibition in Inducing Lung Cancer Cell Death Depend on the Concentration of Cisplatin

doi: 10.1021/acsomega.4c01950

Figure Lengend Snippet: Effect of JNK inhibition on A549 cell viability as a function of cisplatin concentration. (a, b) 7.5 μg/mL concentration of cisplatin becomes lethal in combination with JNK inhibitor SP600125. However, at higher cisplatin concentrations, the inhibitor either protects against cisplatin-induced cell death (a) or does not have any effect (b). (c) Similar effects are observed in the colon cancer cell line DLD-1. (d–g) SP600125 potentiates the appearance of cells with apoptotic morphology in 7.5 μg/mL cisplatin-treated A549 cells (d, e), in contrast to 30 μg/mL cisplatin-treated A549 cells (f, g). Relative cell viability is cell viability after 72 h treatment normalized by initial viability (measured by the MTT method). Representative test results (all measurements were performed in quadruplicate) from more than five experiments are presented. Fluorescent pictures were obtained with a mixture of AO/EB dyes as described in the section. SP—JNK inhibitor SP600125 (20 μM).

Article Snippet: Human non-small-cell lung carcinoma A549 cells were purchased from Cell Lines Service GmbH, Eppelheim, Germany.

Techniques: Inhibition, Concentration Assay

Journal: ACS Omega

Article Title: The Role and Efficacy of JNK Inhibition in Inducing Lung Cancer Cell Death Depend on the Concentration of Cisplatin

doi: 10.1021/acsomega.4c01950

Figure Lengend Snippet: JNK inhibitor SP600125 reduces JNK target transcription factor c-Jun phosphorylation in cisplatin-treated A549 cells. Different concentrations of cisplatin were used. (a) Representative Western blots from 20 and 40 h of treatments are shown. Total protein Coomassie-stained polyacrylamide gels serve as loading controls. (b) Repeated addition (after 24 h) of SP potentiates suppression of c-Jun phosphorylation in response to cisplatin treatment. (c) Repeated addition of SP strengthens the protective effect of JNK inhibition at 15−30 μg/mL concentrations of cisplatin in A549 cells. SP—JNK inhibitor SP600125 (20 μM). p *** < 0.0005, N = 3.

Article Snippet: Human non-small-cell lung carcinoma A549 cells were purchased from Cell Lines Service GmbH, Eppelheim, Germany.

Techniques: Phospho-proteomics, Western Blot, Staining, Inhibition

Journal: ACS Omega

Article Title: The Role and Efficacy of JNK Inhibition in Inducing Lung Cancer Cell Death Depend on the Concentration of Cisplatin

doi: 10.1021/acsomega.4c01950

Figure Lengend Snippet: Cisplatin concentration-dependent effect of different JNK inhibitors on A549 cell viability. Different JNK inhibitors show the same dependence on cisplatin concentration on cell viability. Statistically significant reduction in cell viability is observed at 7.5 μg/mL cisplatin in combination with 10 μM AS601245 (a), 10 μM bentamapimod (b), 2 μM JNK inhibitor IX (c), and 5 μM XG-102 (d). Representative test results (all measurements were performed in quadruplicate) from more than five experiments are presented. (e) Reduction in c-Jun phosphorylation upon the addition of JNK inhibitor XG-102, as determined by Western blot. Representative Western blots are shown. Total protein Coomassie-stained polyacrylamide gels serve as loading controls.

Article Snippet: Human non-small-cell lung carcinoma A549 cells were purchased from Cell Lines Service GmbH, Eppelheim, Germany.

Techniques: Concentration Assay, Phospho-proteomics, Western Blot, Staining

Journal: ACS Omega

Article Title: The Role and Efficacy of JNK Inhibition in Inducing Lung Cancer Cell Death Depend on the Concentration of Cisplatin

doi: 10.1021/acsomega.4c01950

Figure Lengend Snippet: Activation of TP53 in A549 cells in response to different concentrations of cisplatin. (a, b) Expression and phosphorylation at serine-6 is induced by cisplatin and is maximal at 15 μg/mL (6 h of cisplatin treatment). (c, d) Prolonged and increasing expression and phosphorylation of TP53 in cells treated with either 7.5 μg/mL or 30 μg/mL of cisplatin. (e) TP53 activator nutlin-3a potentiates cisplatin-induced decrease in cell viability. (f) Nutlin-3a reduces viability of cisplatin + SP600125-treated cells. C—control without cisplatin; Nut—nutlin-3a (10 μM); SP—SP600125 (20 μM). Representative test results (all measurements were performed in quadruplicate) from more than five experiments are presented, p ** < 0.005, p *** < 0.0005, N = 4. (g) Nutlin induces expression of TP53. Representative Western blots are shown. Total protein Coomassie-stained polyacrylamide gels serve as loading controls.

Article Snippet: Human non-small-cell lung carcinoma A549 cells were purchased from Cell Lines Service GmbH, Eppelheim, Germany.

Techniques: Activation Assay, Expressing, Phospho-proteomics, Control, Western Blot, Staining

Journal: ACS Omega

Article Title: The Role and Efficacy of JNK Inhibition in Inducing Lung Cancer Cell Death Depend on the Concentration of Cisplatin

doi: 10.1021/acsomega.4c01950

Figure Lengend Snippet: Involvement of AKT signaling pathway in cisplatin-induced A549 cell death. (a) AKT protein level does not depend on cisplatin concentration. 6-h-long exposure is presented in the Western blot picture. (b) Phosphorylation/activation of AKT is cisplatin concentration-dependent. 6-h-long exposure is presented in the Western blot picture. (c) AKT protein level does not change during 40 h of cisplatin treatment. (d) Dynamics of AKT activation following cisplatin (7.5 or 30 μg/mL) treatment. (e, f) AKT inhibitor capivasertib enhances cell death at all concentrations of cisplatin used both in the absence (e) and presence (f) of JNK inhibitor SP600125. (g, h) GSK3-β inhibitor SB415286 protects A549 cells from cisplatin both in the absence (g) and presence (h) of JNK inhibition. Representative test results (all measurements were performed in quadruplicate) from more than five experiments are presented. p*<0.05, p**<0.005, p***<0.0005, N = 4. (i) Capivasertib inhibits AKT activity as shown by the inhibition of AKT molecular target GSK3-β phosphorylation at serine-9. Representative Western blots are shown. Total protein Coomassie-stained polyacrylamide gels serve as loading controls. C—control without cisplatin; CAP—capivasertib (10 μM); SP—SP600125 (20 μM), SB—GSK3 inhibitor SB415286 (15 μM).

Article Snippet: Human non-small-cell lung carcinoma A549 cells were purchased from Cell Lines Service GmbH, Eppelheim, Germany.

Techniques: Concentration Assay, Western Blot, Phospho-proteomics, Activation Assay, Inhibition, Activity Assay, Staining, Control

Journal: ACS Omega

Article Title: The Role and Efficacy of JNK Inhibition in Inducing Lung Cancer Cell Death Depend on the Concentration of Cisplatin

doi: 10.1021/acsomega.4c01950

Figure Lengend Snippet: Opposite changes in TP53 and AKT phosphorylation following SP600125 treatment of A549 cells exposed to different concentrations of cisplatin. (a) Expression of TP53 is increased in response to the combination of SP + 7.5 μg/mL cisplatin, in contrast to the combination of SP + 30 μg/mL cisplatin. (b) Phosphorylation of TP53 (serine-6) is increased in response to the combination of SP + 7.5 μg/mL cisplatin, in contrast to the combination of SP + 30 μg/mL cisplatin. (c) Phosphorylation of AKT (threonine-308) is decreased in response to the combination of SP + 7.5 μg/mL cisplatin, in contrast to the combination of SP + 30 μg/mL cisplatin. Representative Western blots are shown. Total protein Coomassie-stained polyacrylamide gels serve as a loading control. SP—JNK inhibitor SP600125 (20 μM); 20 h of treatment.

Article Snippet: Human non-small-cell lung carcinoma A549 cells were purchased from Cell Lines Service GmbH, Eppelheim, Germany.

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

Journal: Bioengineered

Article Title: Knockdown of STIM1 expression inhibits non-small-cell lung cancer cell proliferation in vitro and in nude mouse xenografts

doi: 10.1080/21655979.2019.1669518

Figure Lengend Snippet: Association of STIM1 expression with clinicopathological factors in NSCLC and benign pulmonary diseases.

Article Snippet: Tumor xenografts were generated by subcutaneously inoculating 8x10[ ] A549 cells, Scr-shRNA A549 cells, or STIM1-shRNA A549 cells in Matrigel (cat. no. 354,234, BD Biosciences)[ ].

Techniques: Expressing