slc1a5 inhibitor  (TargetMol)

Journal: Frontiers in Pharmacology

Article Title: Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

doi: 10.3389/fphar.2021.671328

Figure Lengend Snippet: ASCT2 (SLC1A5) protein expression was up-regulated in Almonertinib-induced energy metabolic transport channels. (A) After stimulating H1975 cells with Almonertinib at IC50/2 (4 µM) for 18 h, the cells were collected and LC-MS was used to analysis differences in the expression of related proteins. The arrow shows the glutamine transporter SLC1A5. (B,C) Western blotting (WB) was used to examine the expression of SLC1A5 protein in H1975 cells stimulated by Almonertinib at different times. (D,E) Immunofluorescence was used to detect the expression of SLC1A5 protein in H1975 cells stimulated by Almonertinib at different times.

Article Snippet: After incubation for 24 h, Almonertinib and/or SLC1A5 inhibitor V9302 (MedChemexpress, NJ, United States) at diverse doses (Jiangsu Haoseh Pharmaceutical Group Co., Ltd.) was used to co-incubate H1975 and A549 cells for 24 h. After incubation, CCK-8 assay (Dojin Chemical, Kumamoto, Japan) was conducted to determine cell viability in accordance with specific instructions.

Techniques: Expressing, Liquid Chromatography with Mass Spectroscopy, Western Blot, Immunofluorescence

Journal: Frontiers in Pharmacology

Article Title: Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

doi: 10.3389/fphar.2021.671328

Figure Lengend Snippet: Effects of SLC1A5 inhibition or knockdown on cell proliferation and glutamine uptake in NSCLC cells. (A) CCK8 assay was used to detect different concentrations of V9302 on H1975 and A549 cells for 24, 48 and 72 h. (B) Inhibition SLC1A5 or knockdown of SLC1A5 can reduce the uptake of glutamine in NSCLC cells. Significance: * p < 0.05, ** p < 0.01,*** p < 0.001, ## p < 0.01.

Article Snippet: After incubation for 24 h, Almonertinib and/or SLC1A5 inhibitor V9302 (MedChemexpress, NJ, United States) at diverse doses (Jiangsu Haoseh Pharmaceutical Group Co., Ltd.) was used to co-incubate H1975 and A549 cells for 24 h. After incubation, CCK-8 assay (Dojin Chemical, Kumamoto, Japan) was conducted to determine cell viability in accordance with specific instructions.

Techniques: Inhibition, Knockdown, CCK-8 Assay

Journal: Frontiers in Pharmacology

Article Title: Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

doi: 10.3389/fphar.2021.671328

Figure Lengend Snippet: V9302 enhanced the inhibitory effect of Almonertinib on the proliferation of NSCLC cells. (A) The effect of different concentrations of Almonertinib and 10 µM V9302 on cells alone or in combination was used for 24 h to test the effect of H1975 or A549 cell viability by CCK8. (B) Cell morphological changes observed under an inverted microscope after 12 μM (or 6 μM) Almonertinib and 10 μM V9302 were used separately or in combination for 24 h. (C) Effects of Almonertinib and V9302 alone or in combination on the colony forming ability. A549 cells (A: blank group B: 1 µM V9302 C: 1.2 µM Almonertinib D: 1 µM V9302 and 1.2 µM Almonertinib); H1975 cells (A: blank group B: 1 µM V9302 C: 0.8 µM Almonertinib D: 1 µM V9302 and 0.8 µM Almonertinib). (D) After the intervention of Almonertinib and V9302 alone or in combination for 24 h, the apoptosis rate of A549 cells was detected by double staining. (E) Protein expression of caspase8, caspase3 and PARP. The effects of Almonertinib and V9302 alone or in combination on the expression of A549 or H1975 cells for 24 h were detected by WB. (F) DAPI fluorescence staining was used to observe the nuclear changes of H1975 cells after Almonertinib and V9302 were treated alone or in combination for 24 h. (G) The submicroscopic structure of H1975 cells was observed by electron microscopy after treat with 6 µM Almonertinib combined with 10 µM V9302 for 12 h. Significance: * p < 0.05, ** p < 0.01.

Article Snippet: After incubation for 24 h, Almonertinib and/or SLC1A5 inhibitor V9302 (MedChemexpress, NJ, United States) at diverse doses (Jiangsu Haoseh Pharmaceutical Group Co., Ltd.) was used to co-incubate H1975 and A549 cells for 24 h. After incubation, CCK-8 assay (Dojin Chemical, Kumamoto, Japan) was conducted to determine cell viability in accordance with specific instructions.

Techniques: Inverted Microscopy, Double Staining, Expressing, Fluorescence, Staining, Electron Microscopy

Journal: Frontiers in Pharmacology

Article Title: Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

doi: 10.3389/fphar.2021.671328

Figure Lengend Snippet: V9302 enhanced the antitumor effect of Almonertinib in vivo . (A) Representative tumors from each treatment group. (B) Tumor weight of the mice. (C) Tumor volume of the mice. (D) Evaluation of toxicity in vivo in nude mice. AST, ALT, BUN and Cr of serum were measured by assay kits, respectively. (E) H&E-stained sections of the tumor, liver, lung and kidney from the mice after treatment. (F) TUNEL was used to detect apoptosis in tumor tissues. (G) Protein expression of caspase8, caspase3 and PARP were detected by WB in tumor tissues. Significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After incubation for 24 h, Almonertinib and/or SLC1A5 inhibitor V9302 (MedChemexpress, NJ, United States) at diverse doses (Jiangsu Haoseh Pharmaceutical Group Co., Ltd.) was used to co-incubate H1975 and A549 cells for 24 h. After incubation, CCK-8 assay (Dojin Chemical, Kumamoto, Japan) was conducted to determine cell viability in accordance with specific instructions.

Techniques: In Vivo, Measured Assay, Staining, TUNEL Assay, Expressing

Journal: Frontiers in Pharmacology

Article Title: Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

doi: 10.3389/fphar.2021.671328

Figure Lengend Snippet: Effect of V9302 on Almonertinib-induced autophagy. (A,B) Effect of different drug groups on protein expression of autophagy marker protein LC3-II and P62 in cell lines by WB. (C) Almonertinib alone or combined with chloroquine to observe the effect on the protein level of LC3-II. (D) . Effect of different drug groups on protein expression of autophagy marker protein LC3-II and P62 in tumor tissues. (E) A549 (H1975) cells were treated with 12 μM (6 μM) Almonertinib alone or in combination with 10 μM V9302 for 12 h, then fixed and observed with an electron microscope. (F) mCherry-GFP-LC3B was used to detect the autophagy flux level of Almonertinib combined with V9302 in H1975 cells for 12 h. Significance: * p < 0.05, ** p < 0.01.

Article Snippet: After incubation for 24 h, Almonertinib and/or SLC1A5 inhibitor V9302 (MedChemexpress, NJ, United States) at diverse doses (Jiangsu Haoseh Pharmaceutical Group Co., Ltd.) was used to co-incubate H1975 and A549 cells for 24 h. After incubation, CCK-8 assay (Dojin Chemical, Kumamoto, Japan) was conducted to determine cell viability in accordance with specific instructions.

Techniques: Expressing, Marker, Microscopy

Journal: Frontiers in Pharmacology

Article Title: Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

doi: 10.3389/fphar.2021.671328

Figure Lengend Snippet: CQ increased the proliferation inhibitory effect of Almonertinib combined with V9302 on A549 cells and induces apoptosis. (A) CCK8 assay was used to determine the effect of CQ on the proliferation inhibition of Almonertinib and V9302 co-treated A549 cells. (B,C) Double staining assay was used to detect the effect of CQ on the induction of apoptosis of A549 cells treated with Almonertinib and V9302. Significance: * p < 0.05, ** p < 0.01,*** p < 0.001.

Article Snippet: After incubation for 24 h, Almonertinib and/or SLC1A5 inhibitor V9302 (MedChemexpress, NJ, United States) at diverse doses (Jiangsu Haoseh Pharmaceutical Group Co., Ltd.) was used to co-incubate H1975 and A549 cells for 24 h. After incubation, CCK-8 assay (Dojin Chemical, Kumamoto, Japan) was conducted to determine cell viability in accordance with specific instructions.

Techniques: CCK-8 Assay, Inhibition, Double Staining

Journal: Frontiers in Pharmacology

Article Title: Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

doi: 10.3389/fphar.2021.671328

Figure Lengend Snippet:

Article Snippet: After incubation for 24 h, Almonertinib and/or SLC1A5 inhibitor V9302 (MedChemexpress, NJ, United States) at diverse doses (Jiangsu Haoseh Pharmaceutical Group Co., Ltd.) was used to co-incubate H1975 and A549 cells for 24 h. After incubation, CCK-8 assay (Dojin Chemical, Kumamoto, Japan) was conducted to determine cell viability in accordance with specific instructions.

Techniques: Negative Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TRIM6 Reduces Ferroptosis and Chemosensitivity by Targeting SLC1A5 in Lung Cancer

doi: 10.1155/2023/9808100

Figure Lengend Snippet: TRIM6 modulates ferroptosis via affecting SLC1A5-mediated glutaminolysis. (a) Schematic overview of the glutaminolysis pathway and TCA cycle in ferroptosis. Gln is imported inside the cells by SLC1A5/SLC38A1 and then converted to Glu by GLS in mitochondria. The GOT1 and GLUD1 ultimately converts Glu to α -KG, which contributes to ROS accumulation via the TCA cycle. The small molecule inhibitors are indicated in red: L-Gln transporter inhibitor, GPNA; GLS1 inhibitor, BPTES; GLS inhibitor, 968; pan-transaminase inhibitor, AOA. (b, c) Cell survival and MDA formation in erastin-treated H460 cells ( n = 6). (d, e) Protein levels of SLC1A5 and SLC38A1 in erastin-treated H460 cells with TRIM6 knockdown or overexpression ( n = 6). (f) Relative Gln uptake in erastin-treated H460 cells ( n = 8). (g) Relative SLC1A5 mRNA levels in H460 cells with or without SLC1A5 -OE infection ( n = 6). (h) Relative Gln uptake in erastin-treated H460 cells ( n = 8). (i) Intracellular lipid ROS levels and MDA formation in erastin-treated H460 cells ( n = 6). (j) Cell survival status and colony formation in erastin-treated H460 cells ( n = 6). All data are reported as the mean ± SD, ∗ P < 0.05 versus corresponding groups. NS indicates no significance.

Article Snippet: 2′,7′-dichlorofluorescin diacetate (DCFH-DA, #D6883), superoxide anion assay kit (#CS1000), GSH assay kits (#CS0206), α -KG (#349631), L- γ -glutamyl transpeptidase substrate (SLC1A5 inhibitor; GPNA, #G1135), compound 968 (GLS inhibitor; 968, #352010), bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl) ethyl sulfide (GLS1 inhibitor; BPTES, #SML0601), amino oxyacetate (pan-transaminase inhibitor; AOA, #C13408), cycloheximide (protein synthesis inhibitor; CHX, #01810), MG132 (proteasome inhibitor, #M7449), cisplatin (DDP, #P4394), and paclitaxel (PTX, #1491332) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Over Expression, Infection

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TRIM6 Reduces Ferroptosis and Chemosensitivity by Targeting SLC1A5 in Lung Cancer

doi: 10.1155/2023/9808100

Figure Lengend Snippet: TRIM6 directly interacts with SLC1A5 to promote its degradation. (a) Relative SLC1A5 mRNA levels in erastin-treated H460 cells with or without TRIM6 -OE infection ( n = 6). (b) Protein levels of SLC1A5 in erastin-treated H460 cells after CHX incubation ( n = 6). (c) Ubiquinated levels of SLC1A5 in erastin-treated cells with or without TRIM6 overexpression ( n = 6). (d) Ubiquitination assay in vivo and in vitro ( n = 4). (e) Protein levels of SLC1A5 in erastin-treated H460 cells after MG132 incubation ( n = 6). (f) Relative Gln uptake in erastin-treated H460 cells after MG132 incubation ( n = 6). (g, h) IP assay for examining the interaction between TRIM6 and SLC1A5 ( n = 6). All data are reported as the mean ± SD, ∗ P < 0.05 versus corresponding groups. NS indicates no significance.

Article Snippet: 2′,7′-dichlorofluorescin diacetate (DCFH-DA, #D6883), superoxide anion assay kit (#CS1000), GSH assay kits (#CS0206), α -KG (#349631), L- γ -glutamyl transpeptidase substrate (SLC1A5 inhibitor; GPNA, #G1135), compound 968 (GLS inhibitor; 968, #352010), bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl) ethyl sulfide (GLS1 inhibitor; BPTES, #SML0601), amino oxyacetate (pan-transaminase inhibitor; AOA, #C13408), cycloheximide (protein synthesis inhibitor; CHX, #01810), MG132 (proteasome inhibitor, #M7449), cisplatin (DDP, #P4394), and paclitaxel (PTX, #1491332) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Infection, Incubation, Over Expression, Ubiquitin Assay, In Vivo, In Vitro