Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Design, synthesis and biological evaluation of marine naphthoquinone-naphthol derivatives as potential anticancer agents

doi: 10.1080/14756366.2024.2412865

Figure Lengend Snippet: The antiproliferation effects of compound 5 and 13 . (A) HCT116 and PC9 were treated with the indicated concentrations of compound 5 and 13 (0.01, 0.1, 0.25, 0.5, 1, 2.5, 5, 10 and 20 μM) or DMSO for 72 h. Cell viability was evaluated using CCK-8 assay and shown as relative viability compared to the untreated control. Each test was performed in triplicate. (B) The IC 50 values of compound 5 and 13 in HCT116 and PC9 cells were assessed after 72 h of incubation. (C) Compound 5 and 13 dose-dependently inhibited colony formation in HCT116 cells. Colony formation was assessed after treatment at concentrations of 0.5, 1, 2, and 4 μM for 14 days, and images of crystal violet-stained colonies were depicted. (D) The statistical result of (C). (E) Compound 5 and 13 dose-dependently inhibited colony formation in PC9 cells. Colony formation was assessed after treatment at concentrations of 0.2, 0.5, 1 and 2 μM for 14 d, and images of crystal violet-stained colonies were depicted. (F) The statistical result of (E). All data are shown as mean ± SD, n = 3, t test, * p < .05, ** p < .01, *** p < .001, **** p < .0001.

Article Snippet: The HCT116 and A549 cell lines were acquired from the American Type Culture Collection (ATCC, CCL-247, CCL-185), and the PC9 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC, 90071810).

Techniques: CCK-8 Assay, Control, Incubation, Staining

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Design, synthesis and biological evaluation of marine naphthoquinone-naphthol derivatives as potential anticancer agents

doi: 10.1080/14756366.2024.2412865

Figure Lengend Snippet: Western blot assay of compound 5 and 13 . (A) Western blot assay of EGFR phosphorylation (p-EGFR), PI3K phosphorylation (p-PI3K), Akt phosphorylation (p-Akt), caspase-3, cleaved-caspase-3, and Bcl-2 after treatment of HCT116 cells with 2 μM and 4 μM concentrations of compound 5 and 13 for 24 h. (B) The statistical result of (A). (C) Western blot assay of EGFR phosphorylation (p-EGFR), PI3K phosphorylation (p-PI3K), Akt phosphorylation (p-Akt), caspase-3, cleaved-caspase-3, and Bcl-2 after treatment of PC9 cells with 2 μM and 4 μM concentrations of compound 5 and 13 for 24 h. (D) The statistical result of (C). All data are shown as mean ± SD, n = 3, t test, * p < .05, ** p < .01, *** p < .001, **** p < .0001.

Article Snippet: The HCT116 and A549 cell lines were acquired from the American Type Culture Collection (ATCC, CCL-247, CCL-185), and the PC9 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC, 90071810).

Techniques: Western Blot, Phospho-proteomics

Journal: Antioxidants

Article Title: H 2 S-Synthesizing Enzymes Are Putative Determinants in Lung Cancer Management toward Personalized Medicine

doi: 10.3390/antiox13010051

Figure Lengend Snippet: NSCLC cell lines present cysteine reliance on H 2 S production. ( A ) NSCLC cell lines showed similar basal levels of H 2 S production. To verify if H 2 S production was directly dependent on cysteine degradation, cells were exposed to AOAA/PAG treatment. ( B ) NSCLC cells cultured in the presence of the H 2 S donor NaHS alone and in combination with cysteine showed that H 2 S levels decreased when exposed to AOAA and PAG. Upon cysteine treatment, PC-9 showed a tendency to decrease the average levels of H 2 S in control conditions and compensated production of H 2 S upon exposure to inhibitors. ( C ) Upon cysteine supplementation, NSCLC cell lines tended to decrease H 2 S production at T = 0 h but compensated the CBS and CSE inhibition, maintaining (H292 and PC-9) or increasing (A549) H 2 S levels upon AOAA and PAG exposure. Analysis of H 2 S at T = 0 h indicated that A549 and H292 cell lines presented decreased levels of H 2 S in the presence of cysteine with and without glycolysis inhibition with BPA, while the PC-9 cell line presented no significant changes in all conditions. ( D ) Analysis of H 2 S levels showed that all cell lines presented a similar basal ability to produce H 2 S. Maximum score ( E ) or the average levels ( F ) of ATP production indicated that A549 cells increased the production of ATP upon cysteine supplementation independently of the presence of inhibitors, while H292 and PC-9 were able to maintain ATP levels in all experimental conditions. Maximum ATP score and average levels indicated that A549 cells increased the levels of ATP upon BPA and/or cysteine supplementation, and H292 and PC-9 cells maintained the ATP levels in all culture conditions independently of glycolysis inhibition. All data were normalized to the control condition and are represented as mean ± SD. * p < 0.5, ** p < 0.01, **** p < 0.0001.

Article Snippet: Human adenocarcinoma cell line A549 (CCL-185™), mucoepidermoid carcinoma cell line H292 (CRL-1848™), tubular renal cell line HK2 (CRL-2190™), and keratinocyte cell line HaCaT (PCS-200-011™) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA), and adenocarcinoma cell line PC-9 (90071810) was obtained from European Collection of Authenticated Cell Cultures (ECACC, Porton Down, Salisbury, UK).

Techniques: Cell Culture, Inhibition

Journal: Antioxidants

Article Title: H 2 S-Synthesizing Enzymes Are Putative Determinants in Lung Cancer Management toward Personalized Medicine

doi: 10.3390/antiox13010051

Figure Lengend Snippet: NSCLC cells express distinct expression patterns of enzymes and transporters involved in cysteine metabolism. ( A ) mRNA expression level analysis showed that H292 cells expressed noteworthy levels of CBS and SLC7A11 compared to A549 cells, while PC-9 cells showed low overall expression of CTH , MPST , SLC1A1, and SLC7A11 . ( B ) Immunofluorescence analysis showed that protein expression followed mRNA expression patterns, reporting similar differences. ( C ) mRNA expression level analysis further showed that H292 cells highly expressed GOT2 , while PC-9 showed high expression levels of GOT2 and CDO1 compared to A549 cells. All mRNA expression level data are relative to HPRT1 and represented as mean ± SD. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human adenocarcinoma cell line A549 (CCL-185™), mucoepidermoid carcinoma cell line H292 (CRL-1848™), tubular renal cell line HK2 (CRL-2190™), and keratinocyte cell line HaCaT (PCS-200-011™) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA), and adenocarcinoma cell line PC-9 (90071810) was obtained from European Collection of Authenticated Cell Cultures (ECACC, Porton Down, Salisbury, UK).

Techniques: Expressing, Immunofluorescence

Journal: Antioxidants

Article Title: H 2 S-Synthesizing Enzymes Are Putative Determinants in Lung Cancer Management toward Personalized Medicine

doi: 10.3390/antiox13010051

Figure Lengend Snippet: Distinct molecular backgrounds induce individual metabolic patterns in NSCLC cells. ( A – C ) 1 H-Nuclear magnetic resonance (NMR) of the NSCLC panel studied indicated alterations between A549, H292, and PC-9 regarding levels of intracellular amino acids and peptides, sugars and organic acids, and other metabolites. ( D – F ) NMR of the NSCLC panel studied further indicated alterations between A549, H292, and PC-9 regarding levels of extracellular amino acids and peptides and sugars and organic acids. Data are represented as mean ± SD. * p < 0.5, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( G , H ) PCA showed no differences in the endometabolome ( G ), but the exometabolome ( H ) indicated that A549 and PC-9 cells present distinct metabolic profiles, and H292 cells had common metabolic patterns with A549 and PC-9 cells. ( I ) PLS-DA analysis of the exometabolome allowed the discrimination of the three cell lines. From the PLS-DA analysis of the metabolites present in the cell media, it was possible to discriminate PC-9 from the other cell lines in the first component, while A549 could be discriminated in the second component (upper panel), Q 2 = 0.873. ( J ) The loading plot (lower panel) showing the metabolites important for the discrimination, colored by VIP in the first component. 2-Oxoisocaproate increased in A549, while aspartate, lactate, phosphocholine, and glycerophosphocholine increased in PC-9, making them important for discrimination between these cell lines. ( K ) From the PLS-DA analysis of the metabolites present in the cell media, it was possible to discriminate between A549 and PC-9 cell lines (upper panel), Q 2 = 0.975. ( L ) The loading plot (lower panel) showing the metabolites important for the discrimination, colored by VIP in the first component. ( M ) Metabolites significantly different between PC-9 and A549. Volcano plot (fold change >2 and p value < 0.05) of the intracellular metabolites between A549 and PC-9. Arginine, proline, and nicotinurate were significantly increased in A549 and guanosine in PC-9. ( N ) Nicotinurate was only present in A549 cells. Box plot of nicotinurate intracellular concentrations in the three cell lines (ANOVA analysis p value = 0.00028097 × 10 4 .

Article Snippet: Human adenocarcinoma cell line A549 (CCL-185™), mucoepidermoid carcinoma cell line H292 (CRL-1848™), tubular renal cell line HK2 (CRL-2190™), and keratinocyte cell line HaCaT (PCS-200-011™) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA), and adenocarcinoma cell line PC-9 (90071810) was obtained from European Collection of Authenticated Cell Cultures (ECACC, Porton Down, Salisbury, UK).

Techniques: Nuclear Magnetic Resonance

Journal: Antioxidants

Article Title: H 2 S-Synthesizing Enzymes Are Putative Determinants in Lung Cancer Management toward Personalized Medicine

doi: 10.3390/antiox13010051

Figure Lengend Snippet: NSCLC cell lines are chemoresistant, and SeChry@PURE G4 -FA induces decreased cell viability in NSCLC cells, with specificity toward tumor cells rather than nontumoral cells. ( A ) NSCLC cell lines exposed to the most commonly used therapy regimens showed overall resistance to these drugs, except for cisplatin. ( B ) EC 50 curves and values for SeChry, Sechry@PURE G4- FA, and PURE G4- FA in NSCLC indicated a higher sensitivity of A549 and H292 to the treatment than PC-9. Empty nanoparticles (PURE G4. FA) did not induce relevant toxicity on cell viability. ( C ) EC 50 curves and values for SeChry, Sechry@PURE G4- FA, and PURE G4- FA in noncancer cell lines (HaCaT and HK2) indicated no effect on cell viability. Empty nanoparticles (PURE G4. FA) did not induce relevant toxicity on cell viability. ( D ) Detection of FR-α by immunofluorescence indicated high protein levels in NSCLC but not in nontumoral cell lines. FR- α is labeled in green, and nuclei were counterstained with DAPI (blue). Magnification 400×, scale 20 µm. *** p < 0.001, **** p < 0.0001.

Article Snippet: Human adenocarcinoma cell line A549 (CCL-185™), mucoepidermoid carcinoma cell line H292 (CRL-1848™), tubular renal cell line HK2 (CRL-2190™), and keratinocyte cell line HaCaT (PCS-200-011™) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA), and adenocarcinoma cell line PC-9 (90071810) was obtained from European Collection of Authenticated Cell Cultures (ECACC, Porton Down, Salisbury, UK).

Techniques: Immunofluorescence, Labeling

Journal: Cancer Immunology, Immunotherapy

Article Title: The expression of PD-1 ligand 1 on macrophages and its clinical impacts and mechanisms in lung adenocarcinoma

doi: 10.1007/s00262-022-03187-4

Figure Lengend Snippet: Effect of anti-cancer compounds in cell lines. A schematic diagram of the cell cultures and methods ( A ). A549 and PC9 cells were cultured with dimethyl sulfoxide (DMSO), paclitaxel (PTX), docetaxel (DOC), carboplatin (CBDCA), and pemetrexed (PMET) for 24 h at the same concentration (40 µM), and the mRNA expression of GM-CSF was evaluated by real-time polymerase chain reaction ( B ). The cells were subsequently cultured for another day, and the concentration of GM-CSF in the medium was tested by ELISA ( C ). Macrophages were stimulated with the CM of control PC9 cells or PEMT-treated PC9 cells, and surface PD-L1 expression was detected by fluorescence-activated cell sorting ( D ). Macrophages were stimulated with the CM of PEMT-treated PC9 cells with control immunoglobulin G or anti-GM-CSF antibody (20 µg/mL) (E). *: statistically significant (n = 3 to 4 each), p value < 0.05

Article Snippet: A549 and PC9 were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan).

Techniques: Cell Culture, Concentration Assay, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control, Fluorescence, FACS