human escc cell line kyse30  (Procell Inc)

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 exhibits high expression in ESCC tissues and cells. (A) Sangerbox 3.0 online software assay for PRDX4 expression in pan-cancer. (B) UALCAN database investigation for PRDX4 expression in ESCA samples and normal esophageal epithelial tissues. (C) GEO dataset GSE111011 was used to investigate the expression of PRDX4 in ESCC samples and paired normal samples. (D) RT-qPCR assay was used to assess the expression of PRDX4 in 65 ESCC samples and paired normal samples. (E) Western blot analysis of the protein expression of PRDX4 in eight ESCC samples and paired normal samples. (F) The relative protein levels of PRDX4 in eight ESCC samples and paired normal samples. (G) IHC detection of PRDX4 expression in normal tissues and ESCC tissues. Scale bar, 20 µm. (H) Western blot analysis of PRDX4 protein expression in ESCC cell lines (KYSE70, KYSE450, KYSE520, KYSE30 and KYSE270) and normal esophageal epithelial cell line Het-1A. (I) The relative protein levels of PRDX4 in ESCC cell lines and Het-1A cells. (J) RT-qPCR assay of PRDX4 mRNA expression in the aforementioned ESCC cell lines and Het-1A cells. ** P<0.01, *** P<0.001 and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; ESCA, esophageal carcinoma; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; GEO, Gene Expression Omnibus; IHC, immunohistochemistry; ns, not significant.

Article Snippet: The human ESCC cell line KYSE30 was purchased from Procell Life Science &Technology Co., Ltd. Normal esophageal epithelial cell line Het-1A and human ESCC cell lines KYSE520, KYSE70, KYSE450, and KYSE270, which were authenticated by STR profiling, were purchased from Qingqi Shanghai Biotechnology Development Co., Ltd. ESCC cells were cultured in RPMI-1640 medium (Procell Life Science & Technology Co., Ltd.) supplemented with 10% fetal bovine serum (FBS; Suzhou ShuangRu Biotech Co., Ltd.), as well as penicillin (100 U/ml) and streptomycin (0.1 mg/ml) solution (Solarbio Life Sciences), and were incubated at 37 ̊C with 5% CO 2 .

Techniques: Expressing, Software, Quantitative RT-PCR, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Gene Expression, Immunohistochemistry

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: High expression of PRDX4 predicts a poor prognosis in patients with ESCC (A) UALCAN assay of the effects of PRDX4 expression on the survival of patients with ESCA. (B) GEPIA online software assay of the effects of PRDX4 expression on the survival of patients with ESCA. (C) Sangerbox 3.0 online software assay identifying high PRDX4 expression as a poor prognostic factor in patients with ESCA. (D) RT-qPCR assay for PRDX4 expression in patients with ESCC with different TNM stages. (E) RT-qPCR assay of PRDX4 expression in patients with ESCC without lymph node metastasis and with lymph node metastasis. (F) Log-rank test determination of the prognostic value of PRDX4 in patients with ESCC. ** P<0.01 and *** P<0.001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; ESCA, esophageal carcinoma; GEPIA, Gene Expression Profiling Interactive Analysis; RT-qPCR, reverse transcription quantitative polymerase chain reaction; TNM, tumor-node-metastasis.

Article Snippet: The human ESCC cell line KYSE30 was purchased from Procell Life Science &Technology Co., Ltd. Normal esophageal epithelial cell line Het-1A and human ESCC cell lines KYSE520, KYSE70, KYSE450, and KYSE270, which were authenticated by STR profiling, were purchased from Qingqi Shanghai Biotechnology Development Co., Ltd. ESCC cells were cultured in RPMI-1640 medium (Procell Life Science & Technology Co., Ltd.) supplemented with 10% fetal bovine serum (FBS; Suzhou ShuangRu Biotech Co., Ltd.), as well as penicillin (100 U/ml) and streptomycin (0.1 mg/ml) solution (Solarbio Life Sciences), and were incubated at 37 ̊C with 5% CO 2 .

Techniques: Expressing, Software, Quantitative RT-PCR, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 knockdown suppresses cell proliferation in ESCC cells. (A) Western blot analysis of the protein expression of PRDX4 in KYSE270 cells transfected with PRDX4 siRNA and KYSE30 cells transfected with pcDNA3.1-PRDX4. (B) The relative protein levels of PRDX4 in KYSE270 and KYSE30 cells with different transfections. (C) RT-qPCR assay of the mRNA expression of PRDX4 in KYSE270 cells transfected with PRDX4 siRNA and KYSE30 cells transfected with pcDNA3.1-PRDX4. (D) CCK-8 assay of cell proliferation in KYSE270 cells transfected with PRDX4 siRNA. (E) Colony formation assay of the colony-forming ability of KYSE270 cells transfected with PRDX4 siRNA. (F) Statistical analysis of the number of colonies formed in KYSE270 cells transfected with PRDX4 siRNA. (G) CCK-8 assay of cell proliferation in KYSE30 cells transfected with pcDNA3.1-PRDX4. (H) Colony formation assay of the colony-forming ability of KYSE30 cells transfected with pcDNA3.1-PRDX4. (I) Statistical analysis of the number of colonies formed in KYSE30 cells transfected with pcDNA3.1-PRDX4. (J) EdU staining assay of EdU-positive cells in KYSE270 cells transfected with PRDX4 siRNA. Scale bar, 100 µm. (K) EdU staining assay of EdU-positive cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. Scale bar, 100 µm. (L) Statistical analysis of the number of EdU-positive cells in KYSE270 cells transfected with PRDX4 siRNA. (M) Statistical analysis of the number of EdU-positive cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. *** P<0.001 and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; siRNA, small interfering RNA; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; CCK-8, Cell Counting Kit-8; EdU, 5-ethynyl-2'-deoxyuridine.

Article Snippet: The human ESCC cell line KYSE30 was purchased from Procell Life Science &Technology Co., Ltd. Normal esophageal epithelial cell line Het-1A and human ESCC cell lines KYSE520, KYSE70, KYSE450, and KYSE270, which were authenticated by STR profiling, were purchased from Qingqi Shanghai Biotechnology Development Co., Ltd. ESCC cells were cultured in RPMI-1640 medium (Procell Life Science & Technology Co., Ltd.) supplemented with 10% fetal bovine serum (FBS; Suzhou ShuangRu Biotech Co., Ltd.), as well as penicillin (100 U/ml) and streptomycin (0.1 mg/ml) solution (Solarbio Life Sciences), and were incubated at 37 ̊C with 5% CO 2 .

Techniques: Knockdown, Western Blot, Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Staining, Small Interfering RNA, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 downregulation suppresses cell migration and invasion in ESCC cells. (A) PRDX4 knockdown suppresses cell migration and invasion in KYSE270 cells after transfection with PRDX4 siRNA. Scale bar, 100 µm. (B) Statistical analysis of the number of migratory cells in KYSE270 cells transfected with PRDX4 siRNA. (C) Statistical analysis of the number of invasive cells in KYSE270 cells transfected with PRDX4 siRNA. (D) Western blot analysis of the expression levels of E-cadherin, N-cadherin and vimentin in KYSE270 cells transfected with PRDX4 siRNA. (E) The relative protein levels of E-cadherin, N-cadherin and vimentin in KYSE270 cells transfected with PRDX4 siRNA. (F) PRDX4 overexpression suppresses cell migration and invasion in KYSE30 cells after transfection with pcDNA3.1-PRDX4. Scale bar, 100 µm. (G) Statistical analysis of the number of migratory cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. (H) Statistical analysis of the number of invasive cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. (I) Western blot analysis of the expression levels of E-cadherin, N-cadherin and vimentin in KYSE30 cells transfected with pcDNA3.1-PRDX4. (J) The relative protein levels of E-cadherin, N-cadherin and vimentin in KYSE30 cells transfected with pcDNA3.1-PRDX4. *** P<0.001 and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; siRNA, small interfering RNA.

Article Snippet: The human ESCC cell line KYSE30 was purchased from Procell Life Science &Technology Co., Ltd. Normal esophageal epithelial cell line Het-1A and human ESCC cell lines KYSE520, KYSE70, KYSE450, and KYSE270, which were authenticated by STR profiling, were purchased from Qingqi Shanghai Biotechnology Development Co., Ltd. ESCC cells were cultured in RPMI-1640 medium (Procell Life Science & Technology Co., Ltd.) supplemented with 10% fetal bovine serum (FBS; Suzhou ShuangRu Biotech Co., Ltd.), as well as penicillin (100 U/ml) and streptomycin (0.1 mg/ml) solution (Solarbio Life Sciences), and were incubated at 37 ̊C with 5% CO 2 .

Techniques: Migration, Knockdown, Transfection, Western Blot, Expressing, Over Expression, Small Interfering RNA

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 is an important regulator of ferroptosis in ESCC cells. (A) Determination of MDA, LPO and GSH contents in KYSE270 cells after transfection with PRDX4 siRNA. (B) Western blot analysis of the protein levels of GPX4, SLC7A11 and PTGS2 in KYSE270 cells transfected with PRDX4 siRNA. (C) The relative protein levels of GPX4, SLC7A11 and PTGS2 in KYSE270 cells transfected with PRDX4 siRNA. (D) Determination of MDA, LPO and GSH contents in KYSE30 cells after transfection with pcDNA3.1-PRDX4. (E) Western blot analysis of the protein levels of GPX4, SLC7A11 and PTGS2 in KYSE30 cells transfected with pcDNA3.1-PRDX4. (F) The relative protein levels of GPX4, SLC7A11 and PTGS2 in KYSE30 cells transfected with pcDNA3.1-PRDX4. (G) Detection of the levels of MDA, LPO and GSH in the control group, PRDX4 siRNA group and PRDX4 siRNA plus Fer-1 group in KYSE270 cells. (H) Western blot analysis of the protein expression levels of GPX4, SLC7A11 and PTGS2 in the control group, PRDX4 siRNA group and PRDX4 siRNA plus Fer-1 group in KYSE270 cells. (I) The relative protein levels of GPX4, SLC7A11 and PTGS2 in the control group, PRDX4 siRNA group and PRDX4 siRNA plus Fer-1 group in KYSE270 cells. (J) Detection of the levels of MDA, LPO and GSH in the pcDNA3.1 group, pcDNA3.1-PRDX4 group and pcDNA3.1-PRDX4 plus erastin group in KYSE30 cells. (K) Western blot analysis of the protein expression levels of GPX4, SLC7A11 and PTGS2 in the pcDNA3.1 group, pcDNA3.1-PRDX4 group and pcDNA3.1-PRDX4 plus erastin group in KYSE30 cells. (L) The relative protein levels of GPX4, SLC7A11 and PTGS2 in the pcDNA3.1 group, pcDNA3.1-PRDX4 group and pcDNA3.1-PRDX4 plus erastin group in KYSE30 cells. ** P<0.01, *** P<0.001 and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; siRNA, small interfering RNA; MDA, malondialdehyde; LPO, lipid peroxidation; GSH, glutathione; GPX4, glutathione peroxidase 4; SLC7A11, solute carrier family 7 member 11; PTGS2, prostaglandin-endoperoxide synthase 2; Fer-1, ferrostatin-1; ns, not significant.

Article Snippet: The human ESCC cell line KYSE30 was purchased from Procell Life Science &Technology Co., Ltd. Normal esophageal epithelial cell line Het-1A and human ESCC cell lines KYSE520, KYSE70, KYSE450, and KYSE270, which were authenticated by STR profiling, were purchased from Qingqi Shanghai Biotechnology Development Co., Ltd. ESCC cells were cultured in RPMI-1640 medium (Procell Life Science & Technology Co., Ltd.) supplemented with 10% fetal bovine serum (FBS; Suzhou ShuangRu Biotech Co., Ltd.), as well as penicillin (100 U/ml) and streptomycin (0.1 mg/ml) solution (Solarbio Life Sciences), and were incubated at 37 ̊C with 5% CO 2 .

Techniques: Transfection, Western Blot, Control, Expressing, Small Interfering RNA

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 suppresses ferroptosis of ESCC cells by activating the PI3K/AKT signaling pathway. (A) Determination of MDA, LPO and GSH contents in the absence or presence of the PI3K activator 740 Y-P after PRDX4 knockdown in KYSE270 cells. (B) Western blot analysis of the protein expression levels of GPX4, p-PI3K, PI3K, p-AKT and AKT in the absence or presence of the PI3K activator 740 Y-P after PRDX4 knockdown in KYSE270 cells. (C) The relative protein levels of GPX4, p-PI3K, PI3K, p-AKT and AKT in the absence or presence of the PI3K activator 740 Y-P after PRDX4 knockdown in KYSE270 cells. (D) Detection of MDA, LPO and GSH contents in the absence or presence of the PI3K inhibitor LY294002 after PRDX4 overexpression in KYSE30 cells. (E) Western blot analysis of the protein expression levels of GPX4, p-PI3K, PI3K, p-AKT and AKT in the absence or presence of the PI3K inhibitor LY294002 after PRDX4 overexpression in KYSE30 cells. (F) The relative protein levels of GPX4, p-PI3K, PI3K, p-AKT and AKT in the absence or presence of the PI3K inhibitor LY294002 after PRDX4 overexpression in KYSE30 cells. * P<0.05, ** P<0.01, and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; PI3K, phosphoinositide 3-kinase; AKT, protein kinase B; MDA, malondialdehyde; LPO, lipid peroxidation; GSH, glutathione; GPX4, glutathione peroxidase 4; p-PI3K, phosphorylated PI3K; p-AKT, phosphorylated AKT; ns, not significant.

Article Snippet: The human ESCC cell line KYSE30 was purchased from Procell Life Science &Technology Co., Ltd. Normal esophageal epithelial cell line Het-1A and human ESCC cell lines KYSE520, KYSE70, KYSE450, and KYSE270, which were authenticated by STR profiling, were purchased from Qingqi Shanghai Biotechnology Development Co., Ltd. ESCC cells were cultured in RPMI-1640 medium (Procell Life Science & Technology Co., Ltd.) supplemented with 10% fetal bovine serum (FBS; Suzhou ShuangRu Biotech Co., Ltd.), as well as penicillin (100 U/ml) and streptomycin (0.1 mg/ml) solution (Solarbio Life Sciences), and were incubated at 37 ̊C with 5% CO 2 .

Techniques: Knockdown, Western Blot, Expressing, Over Expression

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: Proposed model of PRDX4-mediated suppression of ferroptosis through regulation of the PI3K/AKT pathway in ESCC. PRDX4 is highly expressed in ESCC samples and cells. High PRDX4 expression is strongly associated with TNM staging and lymph node metastasis in patients with ESCC and may serve as an indicator of prognosis for patients with ESCA. PRDX4 knockdown suppresses cell proliferation and invasion of ESCC cells by inactivating the PI3K/AKT signaling pathway, thereby triggering ferroptosis in these cells. PRDX4, peroxiredoxin 4; PI3K, phosphoinositide 3-kinase; AKT, protein kinase B; ESCC, esophageal squamous cell carcinoma; TNM, tumor-node-metastasis; ESCA, esophageal carcinoma.

Article Snippet: The human ESCC cell line KYSE30 was purchased from Procell Life Science &Technology Co., Ltd. Normal esophageal epithelial cell line Het-1A and human ESCC cell lines KYSE520, KYSE70, KYSE450, and KYSE270, which were authenticated by STR profiling, were purchased from Qingqi Shanghai Biotechnology Development Co., Ltd. ESCC cells were cultured in RPMI-1640 medium (Procell Life Science & Technology Co., Ltd.) supplemented with 10% fetal bovine serum (FBS; Suzhou ShuangRu Biotech Co., Ltd.), as well as penicillin (100 U/ml) and streptomycin (0.1 mg/ml) solution (Solarbio Life Sciences), and were incubated at 37 ̊C with 5% CO 2 .

Techniques: Expressing, Knockdown

Journal: Cancer Biology & Therapy

Article Title: Epigenetic silencing L3MBTL4 sensitizes esophageal cancer to DNA-PKcs inhibitor

doi: 10.1080/15384047.2026.2646393

Figure Lengend Snippet: The expression and methylation status of L3MBTL4 in ESCC cells and tissue samples. (A) The expression of L3MBTL4 in ESCC cells. 5-aza: 5-aza-2'-deoxycytidine; GAPDH: internal control; (–): absence of 5-aza; (+): presence of 5-aza. (B) MSP results in ESCC cells. U: unmethylation alleles; M: methylation alleles; IVD: in vitro methylated DNA, which serves as a methylation control; NL: normal peripheral lymphocytes DNA, which serves as an unmethylation control. (C) Representative MSP results of esophageal intraepithelial neoplasia (EIN). (D) Representative MSP results for esophageal carcinoma (EC) samples. (E) Representative immunohistochemistry staining of L3MBTL4 in esophageal tumor samples and adjacent noncancerous tissue samples. Scale bar: 100 μm (top); 50 μm (bottom). (F) Immunohistochemistry staining score. **** P < 0.0001. (G) Bar diagram indicates an inverse association between the levels of L3MBTL4 and the DNA methylation status. * P < 0.05.

Article Snippet: The human ESCC cell lines KYSE30 (#CL0189), KYSE150 (#CL0493), KYSE180 (#CL0605), KYSE410 (#CL0190), KYSE450 (#CL0606), KYSE510 (#CL0191), and KYSE520 (#CL0913) were acquired from the cell bank of Hunan Fenghui Biotechnology Co., Ltd. COLO680N (#CL0928) was acquired from Procell Company.

Techniques: Expressing, Methylation, Control, In Vitro, Immunohistochemistry, Staining, DNA Methylation Assay

Journal: Cancer Biology & Therapy

Article Title: Epigenetic silencing L3MBTL4 sensitizes esophageal cancer to DNA-PKcs inhibitor

doi: 10.1080/15384047.2026.2646393

Figure Lengend Snippet: Effect of L3MBTL4 on ESCC cell proliferation, colony formation, the cell cycle, and apoptosis. (A) MTT assay showing the effect of L3MBTL4 on ESCC cell proliferation. (B) Effect of L3MBTL4 on colony formation. The scale bar represents the average clone numbers. (C) Bar diagram showing the distribution of the cell phases according to the flow cytometry results. (D) Western blot results of G1/S phase regulators normalized to that of β -actin. n = 3 in each group. (E) Apoptotic results for L3MBTL4 unexpressed and re-expressed cells. (F) Western blot results of apoptotic-related proteins, normalized to β -actin. n = 3 in each group. Vector: empty vector control; L3MBTL4 : L3MBTL4 expressing vector. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The human ESCC cell lines KYSE30 (#CL0189), KYSE150 (#CL0493), KYSE180 (#CL0605), KYSE410 (#CL0190), KYSE450 (#CL0606), KYSE510 (#CL0191), and KYSE520 (#CL0913) were acquired from the cell bank of Hunan Fenghui Biotechnology Co., Ltd. COLO680N (#CL0928) was acquired from Procell Company.

Techniques: MTT Assay, Flow Cytometry, Western Blot, Plasmid Preparation, Control, Expressing

Journal: Cancer Biology & Therapy

Article Title: Epigenetic silencing L3MBTL4 sensitizes esophageal cancer to DNA-PKcs inhibitor

doi: 10.1080/15384047.2026.2646393

Figure Lengend Snippet: L3MBTL4 promotes DNA damage repair in ESCC. (A) IP assay and silver staining. Red arrow: differential band. (B) Co-IP and reciprocal Co-IP results. (C) Immunofluorescence assay showing the colocalization of L3MBTL4 and KU70. (D) IC50 values of cisplatin in L3MBTL4 unexpressed and re-expressed cells. (E) Comet assay in low-dose cisplatin-treated ESCC cells. The data are presented as the mean ± SD of three independent experiments. ** P < 0.01, **** P < 0.0001.

Article Snippet: The human ESCC cell lines KYSE30 (#CL0189), KYSE150 (#CL0493), KYSE180 (#CL0605), KYSE410 (#CL0190), KYSE450 (#CL0606), KYSE510 (#CL0191), and KYSE520 (#CL0913) were acquired from the cell bank of Hunan Fenghui Biotechnology Co., Ltd. COLO680N (#CL0928) was acquired from Procell Company.

Techniques: Silver Staining, Co-Immunoprecipitation Assay, Immunofluorescence, Single Cell Gel Electrophoresis

Journal: Cancer Biology & Therapy

Article Title: Epigenetic silencing L3MBTL4 sensitizes esophageal cancer to DNA-PKcs inhibitor

doi: 10.1080/15384047.2026.2646393

Figure Lengend Snippet: L3MBTL4 activates ATM/CHK2 signaling and inhibits NHEJ signaling. (A) HR and NHEJ efficiencies were evaluated in U2OS-DR-GFP and U2OS-EJ5 cells before and after L3MBTL4 knockdown. (B) HR and NHEJ efficiencies were evaluated in KYSE30 and KYSE150 cells with or without L3MBTL4 expression. (C–E) Levels of ATM/CHK2, ATR/CHK1, and NHEJ signaling in L3MBTL4 unexpressed and re-expressed ESCC cells after treatment with 1 μM cisplatin for 48 h, as detected by immunoblotting, normalized to that of β -actin. n = 3 in each group. (F) The effects of KU70 knockdown on the NHEJ pathways for L3MBTL4 unexpressed and re-expressed ESCC cells normalized to that of β -actin. n = 3 in each group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The human ESCC cell lines KYSE30 (#CL0189), KYSE150 (#CL0493), KYSE180 (#CL0605), KYSE410 (#CL0190), KYSE450 (#CL0606), KYSE510 (#CL0191), and KYSE520 (#CL0913) were acquired from the cell bank of Hunan Fenghui Biotechnology Co., Ltd. COLO680N (#CL0928) was acquired from Procell Company.

Techniques: Knockdown, Expressing, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Epigenetic silencing L3MBTL4 sensitizes esophageal cancer to DNA-PKcs inhibitor

doi: 10.1080/15384047.2026.2646393

Figure Lengend Snippet: Loss of L3MBTL4 expression sensitized ESCC cells to NU7441. (A) IC50 assay showing the sensitivity of ESCC cells to NU7441 under the treatment of 0.5 μM cisplatin. (B) Representative colony formation results showing the synthetic lethal effect of the L3MBTL4 defect and NU7441. (C) Western blot results for NHEJ signaling and γ -H2AX in L3MBTL4 unexpressed and re-expressed cells under the treatment of cisplatin and NU7441 for 48 h, normalized to that of β -actin. n = 3 in each group. Presenting the mean ± SD for three independent experiments. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The human ESCC cell lines KYSE30 (#CL0189), KYSE150 (#CL0493), KYSE180 (#CL0605), KYSE410 (#CL0190), KYSE450 (#CL0606), KYSE510 (#CL0191), and KYSE520 (#CL0913) were acquired from the cell bank of Hunan Fenghui Biotechnology Co., Ltd. COLO680N (#CL0928) was acquired from Procell Company.

Techniques: Expressing, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Epigenetic silencing L3MBTL4 sensitizes esophageal cancer to DNA-PKcs inhibitor

doi: 10.1080/15384047.2026.2646393

Figure Lengend Snippet: Synthetic lethality of L3MBTL4 methylation and DNA-PK inhibitor in vivo . (A) Immunohistochemistry results in L3MBTL4 unexpressed and re-expressed KYSE150 cell xenografts under treatment with cisplatin. (B) L3MBTL4 unexpressed and re-expressed KYSE150 cell xenografts under the treatment with 2 mg/kg cisplatin, 10 mg/kg NU7441 and combination of 2 mg/kg cisplatin with 10 mg/kg NU7441. (C) Growth curves of xenograft tumors. (D) and (E) Normalized tumor volume and weight in L3MBTL4 unexpressed and re-expressed KYSE150 cell xenografts. **** P < 0.0001. (F) A working model for synthetic lethality of L3MBTL4 methylation and DNA-PK inhibitor in ESCC cells.

Article Snippet: The human ESCC cell lines KYSE30 (#CL0189), KYSE150 (#CL0493), KYSE180 (#CL0605), KYSE410 (#CL0190), KYSE450 (#CL0606), KYSE510 (#CL0191), and KYSE520 (#CL0913) were acquired from the cell bank of Hunan Fenghui Biotechnology Co., Ltd. COLO680N (#CL0928) was acquired from Procell Company.

Techniques: Methylation, In Vivo, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: Spatial omics study reveals molecular-cellular dynamics of tumor ecosystem in esophageal squamous-cell carcinoma initiation and progression

doi: 10.1016/j.xcrm.2026.102650

Figure Lengend Snippet: Spatial whole-transcriptome profiling of ESPL and ESCC samples (A) Workflow of DSP experimental process. (B) The UMAP plot displaying the clustering results of AOIs from different compartments at multiple stages of ESCC. (C) The heatmap displaying the expression levels of AOI characteristic markers RNA in different compartments. (D) The boxplots showing the expression levels of marker genes in five different compartments at different stages of ESCC. Box plot shows the median and interquartile range, whiskers extend to 1.5 × IQR. DSP, digital spatial profiling; UMAP, uniform manifold approximation and projection; AOI, area of interest; EP, epithelial-cell-enriched; MC, macrophage-cell-enriched; NC, neutrophil-cell-enriched; ST, stroma-enriched; LS, lymphoid structure; CAFs, cancer-associated fibroblasts; ESPL, esophageal squamous precancerous lesion; non-mESCC, non-metastasis esophageal squamous cell carcinoma; mESCC, metastasis esophageal squamous cell carcinoma; mLN, lymph node metastasis tissues.

Article Snippet: KYSE30 human ESCC cell line , Servicebio , STCC11903P.

Techniques: Expressing, Marker

Journal: Cell Reports Medicine

Article Title: Spatial omics study reveals molecular-cellular dynamics of tumor ecosystem in esophageal squamous-cell carcinoma initiation and progression

doi: 10.1016/j.xcrm.2026.102650

Figure Lengend Snippet: Changes in gene expression patterns in the EP compartment during the initiation of ESCC (A) Volcano plot showed significant DEGs during the formation of ESPL. (B) Pathway enrichment analysis of DEGs in ESPL and normal stages. (C) Changes in gene expression across epidermis development and keratinocyte differentiation pathways during ESPL formation. (D) Interaction plot of ESPL-formation-related genes. Each circle represents a protein, and the interactions are connected by solid lines. The types of interactions were shown on the right side of the figure. (E) The four-quadrant plot illustrates the significant differences in gene expression patterns among four distinct types during the initiation of ESCC. (F) The boxplot shows the expression levels of the presented genes across the four patterns during ESCC initiation. Pattern 1 to 4 were tagged in blue, pink, yellow, and purple, respectively. Box plot shows the interquartile, whiskers extend to1.5 × IQR. (G) Pathway enrichment results for genes in the four patterns. (H) The line graph depicts changes in the signature scores of the indicated pathways during the initiation and development of ESCC. DEGs, differentially expressed genes.

Article Snippet: KYSE30 human ESCC cell line , Servicebio , STCC11903P.

Techniques: Gene Expression, Expressing

Journal: Cell Reports Medicine

Article Title: Spatial omics study reveals molecular-cellular dynamics of tumor ecosystem in esophageal squamous-cell carcinoma initiation and progression

doi: 10.1016/j.xcrm.2026.102650

Figure Lengend Snippet: Spatial transcriptome analysis of the TME during ESCC initiation process (A) Quantitative analysis of immune cells in non-EP compartments during the initiation process of ESCC using TME_consense algorithm. (B) Comparison of B, plasma, CD4 + T, and CD8 + T cells during the initiation process of ESCC. (C) Analyze the immune cell changes in non-EP compartments during the initiation process of ESCC using SpatialDecon algorithm. (D) mIF staining of CD20, CD8, and CD4 on tumor tissues in ESCC initiation stages. Scale bar, 200 μm. (E) Comparison of the percentages of CD20-, CD8-, and CD4-positive cells across the ESCC initiation stages. (F) Pathway enrichment analysis of significantly differential expression genes in non-EP compartments during the initial stage of ESCC. (G) The heatmap shows the differential genes in the MC and NC compartments during the initial stage of ESCC. (H) Comparison of the proportion of tumor-associated macrophage and neutrophile cells across the ESCC development. (I) The heatmap shows the differential genes in TLS compartments during the initial stage of ESCC. (J) mIF staining on TLS compartment of ESCC initiation stage. Scale bar, 100 μm. (K) Comparison of the percentages of APOBEC3A-positive cells between ESPL and non-mESCC stages in the TLS. NE, normal epithelia; ESCC, esophageal squamous cell carcinoma; mIF, multiplex immunofluorescence; MC, macrophage-cell-enriched compartment; NC, neutrophil-cell-enriched compartment; TLS, tertiary lymphoid structures. Box plots show the median and interquartile range, whiskers extend to 1.5×IQR. ∗p < 0.05 , ∗∗ p < 0.01,∗∗∗ p < 0.001, p values were calculated using a two-sided Wilcoxon rank-sum test.

Article Snippet: KYSE30 human ESCC cell line , Servicebio , STCC11903P.

Techniques: Comparison, Clinical Proteomics, Staining, Quantitative Proteomics, Multiplex Assay, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: Spatial omics study reveals molecular-cellular dynamics of tumor ecosystem in esophageal squamous-cell carcinoma initiation and progression

doi: 10.1016/j.xcrm.2026.102650

Figure Lengend Snippet: Characteristic changes in transcription patterns during the progression of ESCC (A) The volcano plot shows significant differences in genes between the early (ESPL and non-mESCC) and advanced (mESCC and mLN) stages of ESCC. (B) Pathway enrichment analysis of differentially expressed genes in the early and advanced stages of ESCC. (C) The gene expression levels associated with significantly enriched pathways throughout the occurrence and development of ESCC. (D) mIF staining of CCND1, LAMB1, and IL-18 on ESCC tissues. Scale bar, 200 μm. (E) Comparison of the percentages of CCND1, LAMB1, and IL-18-positive cells on ESCC tissues. Box plots show the median and interquartile range, whiskers extend to 1.5 × IQR. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, p values were calculated using a two-sided Wilcoxon rank-sum test. (F) The heatmap of gradient-changed DEGs based on pseudotime and ESCC progression of EP.

Article Snippet: KYSE30 human ESCC cell line , Servicebio , STCC11903P.

Techniques: Gene Expression, Staining, Comparison

Journal: Cell Reports Medicine

Article Title: Spatial omics study reveals molecular-cellular dynamics of tumor ecosystem in esophageal squamous-cell carcinoma initiation and progression

doi: 10.1016/j.xcrm.2026.102650

Figure Lengend Snippet: OGT promotes ESCC progression through regulating proliferation, migration, invasion, and apoptosis in vitro and in vivo (A) Representative images of immunohistochemical staining for OGT in cancer and adjacent tissues, magnified at 200× and 400×; scale bars, 100 μm. (B) Paired comparison of OGT IHC scores in tumor and peritumor tissues. p value was calculated using a two-sided Wilcoxon signed-rank test (paired samples, n = 76). (C) Representative WB images of OGT and O-GlcNAc level in all transient transfection groups, including knockdown in KYSE30 and KYSE450 (transfected with siNC, si OGT #1, or si OGT #2) and overexpress in KYSE150 and KYSE410 (transfected with oeVector, oe OGT , or oe OGT and treated with OSMI-1). Images are representative of three independent experiments ( n = 3). (D) OGT and O-GlcNAc level in lentiviral-mediated knockdown KYSE450 and overexpress KYSE150. Representative protein images of three independent experiments were shown ( n = 3). (E and F) Representative images and quantification of plate clone formation assay of lentiviral-mediated knockdown KYSE450 and overexpress KYSE150 cell lines. Data are mean ± SD from three independent experiments ( n = 3). p values were calculated using one-way ANOVA. (G) Cell proliferation measured by the CCK8 assay and relative cell proliferation quantified by OD value at 450 nm of KYSE30, KYSE450, KYSE150, and KYSE410 cell lines. Data are presented as mean ± SD from three independent experiments ( n = 3). p values were calculated using two-way ANOVA with Tukey's multiple-comparisons test. (H) Representative images of cell migration and invasion assays in the KYSE30, KYSE450, KYSE150, and KYSE410 cell lines ( n = 3). Scale bars, 200 μm. (I) Stably transfected shNC/sh OGT KYSE450 cell subcutaneously injected into nude mice. Representative image of xenograft tumors and tumor volume from day 5 to day 30. Data are presented as mean ± SD, n = 5 mice per group. p values were calculated using two-way repeated-measures ANOVA with Sidak's multiple-comparisons test. (J) Bioluminescence imaging of lung metastatic foci at the 7th week in a lung metastasis model. Luciferase activity is measured in photons per cm2 per second per steradian (p/s/cm2/sr). (K) Representative images and quantitative analysis of metastasis nodules on the lung surface. Arrowheads denote the metastasis nodules on the lung surface. (L) Representative images of HE staining of lung metastasis and quantitative analysis of lung metastasis area. Arrowheads denote the metastasis nodules. Scale bars, 200 μm. (J–L) Data are presented as mean ± SD, n = 5 mice per group. p values were calculated using a two-sided Wilcoxon rank-sum test. (M) The results of enriched pathways affected by OGT expression detected across different omics. In the RNA-seq dataset, siOGT cells were defined as the OGT low group, while siNC cells were defined as the OGT-high group. In both the proteomic and glycoproteomic datasets, oeVector cells were defined as the OGT-low group, and oeOGT cells were defined as the OGT-high group. Pathways activated in the OGT-high group are marked in green, while pathways inhibited are marked in red. (N) Comparison of apoptosis rate between KYSE450 siNC and si OGT groups. Data are presented as mean ± SD, n = 3 per group. p values were calculated using unpaired Student's t test. (O) Flow cytometry analysis of cell cycle distribution of KYSE450 cells across siNC and si OGT . ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. NC, negative control group.

Article Snippet: KYSE30 human ESCC cell line , Servicebio , STCC11903P.

Techniques: Migration, In Vitro, In Vivo, Immunohistochemical staining, Staining, Comparison, Transfection, Knockdown, Tube Formation Assay, CCK-8 Assay, Stable Transfection, Injection, Imaging, Luciferase, Activity Assay, Expressing, RNA Sequencing, Flow Cytometry, Negative Control

Journal: Cell Reports Medicine

Article Title: Spatial omics study reveals molecular-cellular dynamics of tumor ecosystem in esophageal squamous-cell carcinoma initiation and progression

doi: 10.1016/j.xcrm.2026.102650

Figure Lengend Snippet: Changes in TME during the progression of ESCC (A) Abundance of 17 cells estimated by SpatialDecon algorithm in non-EP compartments between the early (ESPL and non-mESCC) and advanced stages (mESCC and mLN) of ESCC. (B) Comparison of proportion of presented cells between the early and advanced stages of ESCC. Box plot shows the median and interquartile range, whiskers extend to 1.5 × IQR. p values were calculated using two-sided Wilcoxon rank-sum test. ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (C) Volcano plot showing the DEGs between early and advanced stages in ST compartment. (D) Volcano plot showing the DEGs between early and advanced stages in MC compartment. (E) Volcano plot showing the DEGs between early and advanced stages in NC compartment. (F) Cell-cell interactions based on significant ligand-receptor pairs in advanced stages of ESCC. (G) Interaction relationships of the ACTIVIN, CHEMERIN, and PERIOSTIN pathways among compartments in advanced ESCC. (H) Cell-cell interactions between ST compartments and other compartments via the ACTIVIN, CHEMERIN, and PERIOSTIN pathways during ESCC initiation and progression. (I) The four-quadrant diagram showed the significant differences in gene expression patterns among three different types in the TLS during the ESCC process. Genes showing continuous increase were marked in red, while genes showing continuous decrease were marked in blue. (J) Volcano plot showed significant differentially expressed genes between the TLS of tumor and the lymph follicles of lymph nodules.

Article Snippet: KYSE30 human ESCC cell line , Servicebio , STCC11903P.

Techniques: Comparison, Gene Expression

Journal: Oncology Letters

Article Title: The use of an artificial intelligence algorithm for circulating tumor cell detection in patients with esophageal cancer

doi: 10.3892/ol.2023.13906

Figure Lengend Snippet: (A) Representative images of EpCAM-positive/DAPI-positive cells in patients with esophageal squamous cell carcinoma. Cell nuclei were stained by DAPI and EpCAM is expressed along the plasma membrane. (B) Representative images of DAPI-positive mononuclear cells in healthy volunteers. Cell nuclei were stained with DAPI, but EpCAM is not expressed at the plasma membrane. (C) Number of EpCAM-positive/DAPI-positive cells in healthy volunteers and patients per clinical stage. EpCAM, epithelial cell adhesion molecule.

Article Snippet: Human ESCC cell lines (KYSE30, KYSE140, KYSE520, and KYSE1440) were purchased from the Japanese Collection of Research Bioresources (JCRB, Tokyo, Japan).

Techniques: Staining, Clinical Proteomics, Membrane