kle ec cells  (Servicebio Inc)

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: ATP-based viability assays under DOX treatment. (A) AN3CA cells were exposed to increasing DOX concentrations (0.125–4 µM). The ATPlite assay revealed a dose-dependent reduction in viability, evident from 0.5 µM. (B) KLE cells treated with DOX (0.125–4 µM) displayed a moderate but non–dose-dependent reduction in viability. Data are shown as mean ± SD of six technical replicate wells per condition. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control (medium-only). Vehicle control (DMSO) was included and showed no significant difference vs. untreated control. Experiments were repeated independently twice with consistent results. *p < 0.05; ***p < 0.0005 ****p < 0.0001.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: ATP-based viability assays under menadione treatment. (A) AN3CA cells were exposed to menadione (1–25 µM). The ATPlite assay showed strong cytotoxic effects at 25 µM. (B) KLE cells treated with menadione (1–25 µM) also exhibited markedly reduced viability at 25 µM. Data are shown as mean ± SD of six technical replicate wells per condition. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control (medium-only). Vehicle control (DMSO) was included and showed no significant difference vs. untreated control. Experiments were repeated independently twice with consistent results. ****p < 0.0001.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: ROS generation induced by DOX in EC cell lines. (A) AN3CA cells were exposed to DOX (0.125–4 µM) for 24 h. ROS levels, measured using the CellROX® assay, were significantly elevated at 2 μM and 4 µM. (B) KLE cells treated under the same conditions showed a slight increase in ROS at 2 μM and 4 µM. Data are shown as mean ± SD of six technical replicate wells per condition. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control (medium-only). Vehicle control (DMSO) was included and showed no significant difference vs. untreated control. Experiments were repeated independently twice with consistent results. *p < 0.05; **p < 0.005.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: ROS generation induced by menadione in EC cell lines. (A) AN3CA cells treated with menadione (1–25 µM) for 24 h displayed a significant ROS increase at 10 µM. (B) In KLE cells, ROS elevation remained marginal and did not reach statistical significance across tested doses. Data are shown as mean ± SD of six technical replicate wells per condition. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control (medium-only). Vehicle control (DMSO) was included and showed no significant difference vs. untreated control. Experiments were repeated independently twice with consistent results. *p < 0.05.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: Validation of primary antibodies against SESN2, SESN3, and SOD1 using ICW assay. (A) AN3CA and KLE cells were probed with anti-SESN2 antibody (dilutions 1:1200–1:150). (B) AN3CA and KLE cells were probed with anti-SESN3 antibody (dilutions 1:1000–1:100). (C) AN3CA and KLE cells were probed with anti-SOD1 antibody (dilutions 1:1000–1:100). Secondary antibodies (1:800, LI-COR) were applied, with CellTag™ 520 (1:500, LI-COR) as a loading control for signal normalization. Data are shown as mean ± SD of three technical replicate wells from a single antibody-optimization experiment. Statistical significance was assessed by two-way ANOVA (factors: cell line and antibody dilution), followed by Sidak’s multiple-comparisons test comparing AN3CA vs. KLE within each dilution. **p < 0.005.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Biomarker Discovery, Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: Differential effects of DOX and menadione on SOD1 expression in AN3CA and KLE cells assessed by ICW assay. (A) In AN3CA cells, menadione (1–25 µM) induced an upward trend in normalized SOD1 levels. (B) Raw fluorescence signals corresponding to SOD1 (700 nm) and CellTag™ 520 (520 nm). (C) In KLE cells, menadione increased normalized SOD1 levels at 1–10 μM, followed by a marked decrease at 25 µM. (D) Raw fluorescence signals corresponding to SOD1 (700 nm) and CellTag™ 520 (520 nm). (E) In AN3CA cells treated with DOX (0.125–4 µM), normalized SOD1 levels remained relatively stable at lower concentrations (0.25–0.5 µM) but declined markedly from 1 µM onward. (F) Raw fluorescence signals corresponding to SOD1 (700 nm) and CellTag™ 520 (520 nm). (G) In KLE cells, DOX induced dose-dependent modulation of SOD1 protein levels. (H) Raw fluorescence signals corresponding to SOD1 (700 nm) and CellTag™ 520 (520 nm). Normalized protein levels were calculated as SOD1/CellTag™ 520 ratios (normalized signal). Untreated control (medium-only) wells were included in each experiment and served as the reference group for comparisons. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control. For DOX, the figure shows a representative ICW run (n = 6 technical replicate wells per condition), and the experiment was repeated independently once with consistent results (n = 3). For menadione, data are from one ICW experiment (n = 6 technical replicate wells per condition). *p < 0.05; ***p < 0.0005; ****p < 0.0001.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Expressing, Fluorescence, Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: Differential effects of DOX and menadione on SESN2 expression in AN3CA and KLE cells assessed by ICW assay. (A) In AN3CA cells, menadione (1–25 µM) induced a modest upward trend in normalized SESN2 levels. (B) Raw fluorescence signals corresponding to SESN2 (700 nm) and CellTag™ 520 (520 nm). (C) In KLE cells, menadione increased normalized SESN2 levels at 1–5 μM, followed by a marked decrease at 25 µM. (D) Raw fluorescence signals corresponding to SESN2 (700 nm) and CellTag™ 520 (520 nm). (E) In AN3CA cells treated with DOX (0.125–4 µM), SESN2 exhibited a biphasic response, peaking at 0.125 µM and declining sharply from 1 µM onward. (F) Raw fluorescence signals corresponding to SESN2 (700 nm) and CellTag™ 520 (520 nm). (G) In KLE cells, DOX induced modest SESN2 upregulation at 0.125–0.25 µM, followed by progressive downregulation with the lowest levels at 4 µM. (H) Raw fluorescence signals corresponding to SESN2 (700 nm) and CellTag™ 520 (520 nm). Normalized protein levels were calculated as SESN2/CellTag™ 520 ratios (normalized signal). Untreated control (medium-only) wells were included in each experiment and served as the reference group for comparisons. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control. For DOX, the figure shows a representative ICW run (n = 6 technical replicate wells per condition), and the experiment was repeated independently once with consistent results (n = 3). For menadione, data are from one ICW experiment (n = 6 technical replicate wells per condition). *p < 0.05; **p < 0.005; ***p < 0.0005.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Expressing, Fluorescence, Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: Differential effects of DOX and menadione on SESN3 expression in AN3CA and KLE cells assessed by ICW assay. (A) In KLE cells, menadione (1–25 µM) strongly induced SESN3 in a dose-dependent manner at 1–10 μM, followed by a marked decrease at 25 µM. (B) Raw fluorescence signals corresponding to SESN3 (800 nm) and CellTag™ 520 (520 nm). (C) In AN3CA cells, menadione produced a modest and stable SESN3 response without reaching statistical significance. (D) Raw fluorescence signals corresponding to SESN3 (800 nm) and CellTag™ 520 (520 nm). (E) In AN3CA cells exposed to DOX (0.125–4 µM), SESN3 exhibited a biphasic response, peaking at 0.25 µM and declining sharply from 0.5 µM onward. (F) Raw fluorescence signals corresponding to SESN3 (800 nm) and CellTag™ 520 (520 nm). (G) In KLE cells, DOX induced modest SESN3 upregulation at 0.125–0.25 µM, followed by gradual reduction at higher concentrations. (H) Raw fluorescence signals corresponding to SESN3 (800 nm) and CellTag™ 520 (520 nm). Normalized protein levels were calculated as SESN3/CellTag™ 520 ratios (normalized signal). Untreated control (medium-only) wells were included in each experiment and served as the reference group for comparisons. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control. For DOX, the figure shows a representative ICW run (n = 6 technical replicate wells per condition), and the experiment was repeated independently once with consistent results (n = 3). For menadione, data are from one ICW experiment (n = 6 technical replicate wells per condition). *p < 0.05; **p < 0.005.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Expressing, Fluorescence, Produced, Control

Journal: Frontiers in Physiology

Article Title: Oxidative stress-mediated responses in endometrial cancer cells: contrasting effects of doxorubicin and menadione

doi: 10.3389/fphys.2026.1733194

Figure Lengend Snippet: SESN2, SESN3, and SOD1 gene expression profiles in AN3CA and KLE cells analyzed by qRT-PCR. (A) SESN2 mRNA was significantly upregulated in both cell lines following 4 µM DOX treatment. Menadione induced robust SESN2 upregulation at 25 µM in KLE cells. (B) SESN3 mRNA was strongly upregulated by DOX in both lines, whereas menadione downregulated SESN3 in both cell lines. (C) SOD1 mRNA showed significant upregulation in KLE cells under 4 µM DOX, while AN3CA cells exhibited no significant changes. Under menadione, AN3CA cells displayed significant downregulation of SOD1 at 10 µM and 25 μM, while KLE cells showed downregulation at 25 µM. RT–qPCR data were normalized to UBC as the endogenous reference gene and are presented relative to the untreated control (medium-only). Each condition was analyzed in three technical replicate wells. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test comparing each treatment condition to the untreated control. Key findings were confirmed in an independent repeat experiment. *p < 0.05; **p < 0.005; ***p < 0.0005.

Article Snippet: The EC cell lines KLE (CRL-1622TM) and AN3CA (HTB-111TM) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Gene Expression, Quantitative RT-PCR, Control

Journal: Discover Oncology

Article Title: PTBP1 functions as a suppressor of ferroptosis in endometrial carcinoma cells by stabilizing SLC7A11 mRNA

doi: 10.1007/s12672-025-04128-0

Figure Lengend Snippet: In vitro functional analysis of PTBP1 in endometrial cancer cells and in vivo validation using an Ishikawa xenograft model. A and B Transfection with shRNA knockdown vectors targeting PTBP1 significantly reduced both mRNA and protein levels in Ishikawa and KLE cell lines. C PTBP1 knockdown notably decreased cell viability in both Ishikawa and KLE cells. D – H Cell-derived xenograft (CDX) experiments in mice implanted with Ishikawa cells demonstrated that PTBP1 knockdown significantly reduced tumor volume and weight. Subsequent IHC staining of the xenograft tissues revealed a substantial decrease in Ki-67 protein levels and increase in 4-HNE level upon PTBP1 knockdown. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Ishikawa EC cells (#CL-0823, Procell, Wuhan, China) and KLE EC cells (#STCC10610P, Servicebio, Wuhan, China) were cultured at 37 °C in Dulbecco’s Modified Eagle Medium (DMEM , Procell) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin-penicillin, in a 5% CO 2 atmosphere.

Techniques: In Vitro, Functional Assay, In Vivo, Biomarker Discovery, Transfection, shRNA, Knockdown, Derivative Assay, Immunohistochemistry

Journal: Discover Oncology

Article Title: PTBP1 functions as a suppressor of ferroptosis in endometrial carcinoma cells by stabilizing SLC7A11 mRNA

doi: 10.1007/s12672-025-04128-0

Figure Lengend Snippet: PTBP1 knockdown facilitates ferroptosis in endometrial cancer cells. A Measurement of ROS production using DCF-DA probes. B GSH, C MDA, and D Fe 2+ levels in Ishikawa and KLE EC cells transfected with PTBP1-shRNA (shPTBP1) or nontargeting control (shNC) vectors, measured by relevant assay kits. E– I Ishikawa and KLE EC cells transfected with shPTBP1 or shNC for 48 h were analyzed for mRNA expression of ACSL4 ( E ) and GPX4 ( F ), by RT-qPCR, as well as protein expression of ACSL4 ( G and H ) by Western blot, and GPX4 ( I ) by immunofluorescence. J and K The mRNA expression of SLC7A11 ( J ), as well as protein expression of SLC7A11, ACSL4, and GPX4 ( K ) in mice tumor tissues were determined by RT-qPCR and western blot assays. L The protein level of HSP27 in mice tumor tissues was tested by IHC. M Cell viability of Ishikawa and KLE cells transduced with shNC or shPTBP1 and treated with DMSO, Erastin (10 µM), or Erastin plus Ferrostatin-1 (Fer-1, 1 µM) for 24 h, measured by CCK-8 assay. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Ishikawa EC cells (#CL-0823, Procell, Wuhan, China) and KLE EC cells (#STCC10610P, Servicebio, Wuhan, China) were cultured at 37 °C in Dulbecco’s Modified Eagle Medium (DMEM , Procell) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin-penicillin, in a 5% CO 2 atmosphere.

Techniques: Knockdown, Transfection, shRNA, Control, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Transduction, CCK-8 Assay

Journal: Discover Oncology

Article Title: PTBP1 functions as a suppressor of ferroptosis in endometrial carcinoma cells by stabilizing SLC7A11 mRNA

doi: 10.1007/s12672-025-04128-0

Figure Lengend Snippet: PTBP1 mediates SLC7A11 mRNA stability by binding to its 5’UTR. A Venn diagram displaying the overlap between differentially expressed genes (DEGs) (using criteria of p -adj < 0.05 and |log 2 FoldChange| >0.58) in PTBP1-KD Ishikawa EC cells versus shNC controls, ferroptosis-related genes, and predicted PTBP1 targets identified by the StarBase algorithm. B Transcriptomic analysis of TCGA revealed the mRNA expression levels of SLC7A11 and CBS in EC tissues compared to normal tissues. C Quantitative PCR analysis showing enrichment of SLC7A11 mRNA in anti-PTBP1 immunoprecipitates compared to anti-IgG controls. D Schematic representation of the SLC7A11 5’UTR, the predicted PTBP1 binding site (WT), and the mutated binding site (MUT). E Relative luciferase activity of WT or MUT reporters in shPTBP1- or shNC-transfected 293T cells. F DNA pull down-western blot showing PTBP1 enrichment in Bio-SLC7A11 5’UTR pull-downs compared to Bio-NC controls. G and H Quantification of SLC7A11 mRNA in shPTBP1- or shNC-transfected Ishikawa and KLE cells. I and J mRNA stability analysis using the actinomycin D exposure method, with RNA collected at 0, 1, 2, 3, and 4 h after exposure and quantified by PCR. K and L Measurement of SLC7A11 protein levels in PTBP1 KD and control Ishikawa and KLE cells. * p < 0.05, ** p < 0.01, *** p < 0.001, n.s. non-significant

Article Snippet: Ishikawa EC cells (#CL-0823, Procell, Wuhan, China) and KLE EC cells (#STCC10610P, Servicebio, Wuhan, China) were cultured at 37 °C in Dulbecco’s Modified Eagle Medium (DMEM , Procell) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin-penicillin, in a 5% CO 2 atmosphere.

Techniques: Binding Assay, Expressing, Real-time Polymerase Chain Reaction, Luciferase, Activity Assay, Transfection, Western Blot, Control

Journal: Discover Oncology

Article Title: PTBP1 functions as a suppressor of ferroptosis in endometrial carcinoma cells by stabilizing SLC7A11 mRNA

doi: 10.1007/s12672-025-04128-0

Figure Lengend Snippet: PTBP1 disruption induces ferroptosis through downregulation of SLC7A11. A – F Western blot analysis of SLC7A11 protein ( A ), CCK8 assay for cell viability ( B ), ROS production measured using DCFH-DA probes and fluorescence analysis ( C ), GSH content ( D ), MDA expression ( E ), and Fe2 + levels ( F ) in Ishikawa and KLE EC cells transfected with shPTBP1, shPTBP1 + SLC7A11-ov, shPTBP1 + vec, or nontargeting control shNC. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Ishikawa EC cells (#CL-0823, Procell, Wuhan, China) and KLE EC cells (#STCC10610P, Servicebio, Wuhan, China) were cultured at 37 °C in Dulbecco’s Modified Eagle Medium (DMEM , Procell) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin-penicillin, in a 5% CO 2 atmosphere.

Techniques: Disruption, Western Blot, CCK-8 Assay, Fluorescence, Expressing, Transfection, Control

Journal: Discover Oncology

Article Title: PTBP1 functions as a suppressor of ferroptosis in endometrial carcinoma cells by stabilizing SLC7A11 mRNA

doi: 10.1007/s12672-025-04128-0

Figure Lengend Snippet: Knockdown of SLC7A11 alone is sufficient to induce ferroptosis in endometrial cancer cells. A Western blot analysis validating the knockdown efficiency of SLC7A11 in Ishikawa and KLE cells transfected with shSLC7A11 or shNC. B Cell viability measured by CCK-8 assay in Ishikawa and KLE cells after SLC7A11 knockdown . C and D Intracellular ROS levels detected by DCFH-DA fluorescence in Ishikawa and KLE cells after SLC7A11 knockdown. E – G Assessment of key ferroptosis biochemical markers in Ishikawa and KLE cells after SLC7A11 knockdown, including E glutathione (GSH) levels,​ F ​ malondialdehyde (MDA) content, and G ferrous iron (Fe 2+ ) concentration. ** p < 0.01, *** p < 0.001

Article Snippet: Ishikawa EC cells (#CL-0823, Procell, Wuhan, China) and KLE EC cells (#STCC10610P, Servicebio, Wuhan, China) were cultured at 37 °C in Dulbecco’s Modified Eagle Medium (DMEM , Procell) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin-penicillin, in a 5% CO 2 atmosphere.

Techniques: Knockdown, Western Blot, Transfection, CCK-8 Assay, Fluorescence, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Pan-cancer multi-omics characterization of calcyphosine and its revealed links to the immune microenvironment and regulatory networks in endometrial carcinoma

doi: 10.3389/fimmu.2025.1688606

Figure Lengend Snippet: CAPS expression in EC and its impact on cell proliferation. (A) Western blot analysis of CAPS protein levels in EC tissues and paired adjacent non-tumor tissues. (B) Quantification of CAPS protein expression based on Western blot results. (C) Representative immunohistochemical (IHC) staining images of CAPS in EC and adjacent tissues at 20× and 40× magnification. (D) Statistical analysis of IHC scores (n = 12 per group). (E, F) CCK-8 assays evaluating cell viability in KLE and Ishikawa cells following CAPS knockdown. (G, H) EdU staining assays assessing proliferative capacity after CAPS knockdown in KLE and Ishikawa cells. (I, J) Quantification of EdU-positive cells. (K) CAPS knockdown efficiency validated by Western blot in Ishikawa and KLE cells. CAPS protein levels were markedly reduced after transfection with Si1-CAPS and Si3-CAPS compared with Si-NC. β-actin served as the loading control.(* P < 0.05, ** P < 0.01, **** P < 0.0001).

Article Snippet: The human EC cell lines KLE and Ishikawa were obtained from GeneChem Co., Ltd. (Shanghai, China).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Immunohistochemistry, CCK-8 Assay, Knockdown, Staining, Transfection, Control

Journal: Frontiers in Immunology

Article Title: Pan-cancer multi-omics characterization of calcyphosine and its revealed links to the immune microenvironment and regulatory networks in endometrial carcinoma

doi: 10.3389/fimmu.2025.1688606

Figure Lengend Snippet: CAPS knockdown promotes migration and invasion of EC cells. (A, B) Wound healing assays evaluating the effect of CAPS knockdown on wound closure in KLE and Ishikawa cells. (C, D) . Quantification of wound healing areas. (E, F) Transwell assays assessing the impact of CAPS knockdown on cell migration (E) and invasion (F) in KLE and Ishikawa cells. (G, H) Quantification of migrated (G) and invaded (H) cells in Transwell assays. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: The human EC cell lines KLE and Ishikawa were obtained from GeneChem Co., Ltd. (Shanghai, China).

Techniques: Knockdown, Migration