Journal: Discover Oncology

Article Title: Lipid metabolism-related genes correlate with immune microenvironment and regulate the efficacy of immunotherapy via ferroptosis in melanoma

doi: 10.1007/s12672-025-04163-x

Figure Lengend Snippet: Screening and functional validation of differentially-expressed ferroptosis-related genes. (A) Venn diagrams of the differentially-expressed ferroptosis-related genes that have prognostic potential. (B) The expression of ALOX5, ACSL4, ACACA, ABCC1 in the two subgroups in training cohort, as well as Kaplan-Meier analysis of the survival of patients in the two subgroups. (C) The knockdown efficiency of ASCL4 and ALOX5 in melanoma cells, as well as cell viability of melanoma cells in response to RSL3 treatment. (D) Lipid ROS levels of indicated cells measured by flow cytometry using C11-BODIPY. Data represent the mean ± SEM of triplicates. P value was calculated by two-tailed Student’s t-test and log-rank tests. ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns, non-significant. n (cluster 1) = 208, n (cluster 2) = 246. n (low) = 227, n (high) = 227

Article Snippet: The primary antibodies and dilutions for western blotting (WB) and immunofluorescence (IF) staining analysis are listed below: ACSL4 (22401-1-AP, Proteintech, Wuhan, China; 1:100 for IHC, 1:100 for IF and 1:5 000 for WB), ALOX5 (10021-1-Ig, Proteintech; 1:400 for IHC, and 1:500 for WB), PTGS2 (66351-1-Ig, Proteintech; 1:400 for IF), CD8α(ab22378,, Abcam; 1:300 for IF), ACTIN (66009-1-Ig, Proteintech; 1:10 000 for WB).

Techniques: Functional Assay, Biomarker Discovery, Expressing, Knockdown, Flow Cytometry, Two Tailed Test

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 1 Alox5-5-HETE axis is upregulated in gastric cancer tissues. (A) Scatter plot of Alox5 protein level in paired normal and tumor tissues obtained from 52 patients with gastric tumor. (B) Representative immunohistochemistry analysis of normal and tumor gastric tissues from patient#23 performed by Alox5 staining. (C) Scatter plot of 5-HETE level in paired normal and tumor tissues obtained from 52 patients with gastric tumor. Alox5 and 5-HETE were assessed using tissue homogenates and quantified using ELISA assay. (D) Relative tumor/normal ratio value of Alox5 and 5-HETE levels in individual gastric cancer patients. Alox5 or 5-HETE level in normal tissues were set as 1 (indicated by a red line)

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 2 ALOX5 overexpression promotes cell growth and protects gastric cancer cells from chemotherapeutic agents-induced toxicity. (A and B) Analysis of proliferation of AGS and N87 cells after ALOX5 overexpression performed by BrdU labeling. (C and D) Analysis of colony formation of AGS and N87 cells after ALOX5 overexpression. (E and F) Analysis of migration of AGS and N87 cells after ALOX5 overexpression. Analysis of proliferation (G and H) and apoptosis (I and J) after treatment of 5-FU and cisplatin in ALOX5-overexpressing AGS and N87 cells. Results are presented as relative to control. Proliferation and apoptosis assays were assessed after 72 h of drug treatment. 5-FU at 200 nM and cisplatin at 300 nM were used. *p < 0.05, compared to p-Vector. #p < 0.05, compared to cisplatin or 5-FU alone

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Over Expression, Labeling, Migration, Control, Plasmid Preparation

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 4 ALOX5 knockdown inhibits gastric cancer and enhances the toxicity of chemotherapeutic agents. Analysis of proliferation (A and B) and apoptosis (C and D) in AGS and N87 cells after ALOX5 knockdown, and in the presence of 5-FU and cisplatin. ALOX5 knockdown significantly enhances the anti-proliferative and pro-apoptotic effects of 5-FU and cisplatin in gastric cancer cells. 5-FU at 50 nM and cisplatin at 50 nM were used. Proliferation and apoptosis assays were assessed after 72 h of drug treatment. Results are presented as relative to control. *p < 0.05, compared to control. #p < 0.05, compared to cisplatin or 5-FU

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Knockdown, Control

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 6 ALOX5 knockdown inhibits ERK in gastric cancer cells. Representative western blot photo (A) and quantification analysis by Image J (B) of p-Erk, p-p90RSK, p-Akt, pol II S5, Mcl-1, Bim, and Bcl levels in N87 cells after ALOX5 knockdown. Scr siRNA value was set as 1 and indicated as line. *p < 0.05, compared to Scr siRNA

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Knockdown, Western Blot

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 5 Alox5 inhibitors suppress gastric cancer and enhance the toxicity of chemotherapeutic agents. (A and B) Proliferation level of AGS and N87 cells after zileuton and AA861 treatment in the presence of 5-FU and cisplatin, as evaluated by BrdU labeling. (C and D) Proliferation level of AGS and N87 cells after zileuton and AA861 treatment in the presence of 5-FU and cisplatin, as evaluated by TUNEL assay. Zileuton and AA861 significantly enhance the anti-proliferative and pro-apoptotic effects of 5-FU and cisplatin in gastric cancer cells. 5-FU at 50 nM and cisplatin at 50 nM were used. Zileuton at 100 μM and 200 μM, AA86 at 30 μM and 60 μM were used in proliferation and apoptosis assays, respectively. Proliferation and apoptosis assays were assessed after 72 h of drug treatment. *p < 0.05, compared to control. #p < 0.05, compared to cisplatin or 5-FU

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Labeling, TUNEL Assay, Control

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 7 ALOX5-5-HETE axis activates ERK in gastric cancer cells. (A) Representative western blot photo of p-Erk, p-p90RSK, p- Akt, Mcl-1, Bim, and Bcl-2 in N87 cells after ALOX5 overexpression or 5-HETE addition. (B) Proliferation level of treatment of U0126 and LY2780301 in ALOX5-overexpressing N87 cells. (C) Proliferation level of treatment of U0126 and LY2780301 in N87 cells in the presence of 5-HETE. U0126 (MEK inhibitor, 10 μM) but not LY2780301 (Akt inhibitor, 10 μM) significantly reveres the pro-proliferative effect induced by ALOX5 overexpression and addition of 5-HETE. Inhibitors were added to the cells at 48h post-transfection. *p < 0.05, compared to control; ns, not significant

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Western Blot, Over Expression, Transfection, Control