Journal: Nature Communications

Article Title: Intracellular Fusobacterium nucleatum infection attenuates antitumor immunity in esophageal squamous cell carcinoma

doi: 10.1038/s41467-023-40987-3

Figure Lengend Snippet: a Three-dimensional visualization of α-tubulin and Fn in E109 cells. The cells were infected with Fn (MOI of 1:10) for 48 h. Scale bar: 50 μm (left). b , c Intracellular bacterial proliferation was assessed by a gentamycin protection assay. E109, Kyse150 and AKR cells were lysed at the indicated time points after Fn (MOI of 1:10) infection, and the numbers of total viable bacteria ( b ) and viable bacteria per ESCC cell ( c ) were determined by the serial dilution method. P means vs . the 24 h group. d , e FACS of PD-L1 + membrane expression in E109, Kyse150 and AKR cells after infection with Fn (MOI of 1:10) for 48 h and quantification. f , g Dichromatic IF staining of PD-L1 and Fn in E109, Kyse150 and AKR cells. The cells were infected with Fn (MOI of 1:10) for 48 h. f Representative images. Scale bar: 20 μm. g Quantification. h qRT‒PCR analysis of PD-L1 in E109, Kyse150 and AKR cells after infection with Fn (MOI of 1:1, 1:10, 1:100) for 48 h or infection with Fn (MOI of 1:10) for 24, 48 or 72 h. P means vs . the Con group. i , j Immunoblotting analysis of PD-L1 in E109, Kyse150 and AKR cells after infection with Fn ( i ) and quantification ( j ). P means vs . the Con group. k Immunoblotting analysis and quantification of PD-L1 in tumors from C57BL/6 xenografts (mean ± SD; the experiment was done once; n = 3 mice per group). l IF staining of PD-L1 or Fn in tumor tissues from C57BL/6 xenografts. Scale bar: 50 μm. Images in a and l were representative results of n = 3 independent experiments with similar results. Results in b , c , e , g , h and j were presented as n = 3 biological replicates, mean ± SD, ns means not significant. The statistical significance of results in b , c , h , j and k were determined by one-way ANOVA analysis. e and g were determined by a two-tailed unpaired t -test.

Article Snippet: The human ESCC cell line Eca109 (E109) and Kyse150 (a kind gift from professor Musheng Zeng, SYSUCC, Guangzhou, China), Jurkat cell line (ATCC TIB-152), embryonic kidney 293 T cells (ATCC CRL-3216), and mouse ESCC cell line AKR (C945, Wuhan Sunncell Biotechnology, China) were cultured in RPMI 1640 or DMEM medium (Gibco, CA, USA), respectively.

Techniques: Infection, Bacteria, Serial Dilution, Membrane, Expressing, Staining, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Intracellular Fusobacterium nucleatum infection attenuates antitumor immunity in esophageal squamous cell carcinoma

doi: 10.1038/s41467-023-40987-3

Figure Lengend Snippet: a – c Dichromatic IF staining of Fn and Fn-Dps ( a ), scale bar: 40 μm (top). Immunoblotting ( b ) and FACS ( c ) analysis of PD-L1. Cells were infected with Fn (MOI of 1:10) or treated with Fn-Dps (1 μM) for 48 h. d – f Venn diagram showing the unique and overlapping predicted transcription factors in different groups. The blue dots represent differentially expressed transcription factor genes (POU6F1, CEBPB, NR4A2 and MAFG). qRT‒PCR analysis ( f ). g Analysis of the relevance of the CD274 (PD-L1) and ATF3 genes in ESCA tissues (TCGA database, n = 173). h – j ATF3 and PD-L1 expression was analyzed by western blotting. ESCC cells were transfected with ATF3 overexpression (OE) or negative control (CTRL) vectors for 48 h ( h ). Co-IP assay ( i ). Cells were treated with Fn-Dps (1 μM) for 48 h after transfection with siRNA against ATF3 for 48 h ( j ). k , l Representative images of IHC staining ( k ) and IF staining quantification ( l ) of ATF3 or PD-L1 expression in tumors from C57BL/6 xenografts. Scale bar: 10 μm. m The interaction of Fn-Dps with ATF3 in ESCC cells after treated with Fn-Dps (1 μM) for 48 h was detected by Co-IP assay. n Analysis of CD274 WT or mutant promoter activity in 293 T cells transfected with ATF3-Flag and treated with Fn-Dps (1 μM) for 48 h. o ChIP‒qPCR analysis of the relative enrichment of ATF3 at the CD274 gene promoter in Kyse150 cells. p , q Dichromatic IF staining of ATF3 and Fn-Dps (p), scale bar: 50 μm (top). Immunoblotting of nuclear (N) and cytoplasmic (C) ( q ). Cells were treated with Fn-Dps (1 μM) for 48 h. Images in a , h , j , m , p and q were representative results of n = 3 independent experiments with similar results. Results in b , c , n , and o were presented as n = 3 biological replicates, mean ± SD. Statistical significance in b , c was determined by a two-tailed unpaired t -test. l , n and o were determined by two-way ANOVA multiple comparisons. g was determined by two-tailed nonparametric Spearman correlation analysis.

Article Snippet: The human ESCC cell line Eca109 (E109) and Kyse150 (a kind gift from professor Musheng Zeng, SYSUCC, Guangzhou, China), Jurkat cell line (ATCC TIB-152), embryonic kidney 293 T cells (ATCC CRL-3216), and mouse ESCC cell line AKR (C945, Wuhan Sunncell Biotechnology, China) were cultured in RPMI 1640 or DMEM medium (Gibco, CA, USA), respectively.

Techniques: Staining, Western Blot, Infection, Expressing, Transfection, Over Expression, Negative Control, Co-Immunoprecipitation Assay, Immunohistochemistry, Mutagenesis, Activity Assay, Two Tailed Test

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: LIMK1 as a Novel Kinase of β-Catenin Promotes Esophageal Cancer Metastasis by Cooperating With CDK5.

doi: 10.1002/advs.202503223

Figure Lengend Snippet: Figure 7. The combination therapy targeting LIMK1 and CDK5 significantly inhibits ESCC metastasis. A) Transwell assays were performed to determine the invasive abilities of EAC, gastric cancer, and lung cancer cells when treated with BMS-5 and/or Dinaciclib. B) The flow diagram shows the lung metastasis model, lymph node metastasis model in mice, and the treatment plan. C) Bioluminescence imaging and quantification of lung metastasis in mice intravenously injected with ESCC cells and treated with BMS-5 and/or Dinaciclib. Hematoxylin-eosin (H&E) staining shows lung metastasis as indi- cated. D) The lymph node metastasis model was established and the therapeutic effects of BMS-5 and/or Dinaciclib were monitored by Bioluminescence imaging. E) Key indicators of drug toxicity were detected in mice treated with indicated inhibitor or DMSO.

Article Snippet: Cell Culture and Drugs: Human ESCC cell lines KYSE150 and KYSE410 were purchased fromDeutsche Sammlung vonMikroorganismen undZellkulturen (DSMZ, Braunschweig, Germany).

Techniques: Imaging, Injection, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Construction of a risk signature using the least absolute shrinkage and selection operator (LASSO) analysis. Partial likelihood deviances for (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Coefficient profiles of senescence‐related gene pairs for (C) ESCC and (D) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Selection

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Kaplan–Meier analysis of high‐ and low‐risk patients (red and blue, respectively) with (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Receiver operating characteristic curves for (C) ESCC and (D) EAC. Survival risk curves (top) and sand scatter plots (bottom) for (E) ESCC and (F) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Correlations of immune microenvironments evaluated using ESTIMATE. (A) Immune score and (B) ESTIMATE score for oesophageal squamous cell carcinoma (ESCC). (C) Immune score and (D) ESTIMATE score for oesophageal adenocarcinoma (EAC). Relationships between risk and immune scores for (E) ESCC and (H) EAC. Relationships between risk and ESTIMATE scores for (F) ESCC and (G) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Random forest error rates (left graphs) and relative importance (right graphs) for (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Expression of IRF5 and BMI1 in (C) ESCC, (D) EAC and (E) EC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Expression analyses of IRF5 in four ESCC cell lines using western blotting (A, B). The efficiency of IRF5 ‐knockdown in KYSE150 cells was determined using western blotting (C, D). Ctrl: No siRNA infection; NC: Negative control. Statistical analyses of n = 3 independent experiments were assessed. Results are shown as mean ± SD, ns p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Expressing, Western Blot, Knockdown, Infection, Negative Control

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: a) Preparation of aptamer EA1‐modified PTX/siEFNA1‐loaded nanomedicine. b) Schematic diagram of the proposed EA1‐EYLNs‐PTX/siEFNA1 by the modification of EYLNs with ESCC specific aptamer EA1 and co‐loading with PTX/siEFNA1 for enhanced therapeutic effect against ESCC.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: Modification

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: EFNA1 is overexpressed in esophageal squamous cell carcinoma with pulmonary metastasis and correlates with poor patient prognosis. a) Differentially expressed molecules in ESCC cancer and adjacent tissues using protein mass spectrographic analysis. b) EFNA1 mRNA expression in TCGA RNA‑seq database. c) Overall survival of patients displaying high or low/middle EFNA1 expression using GEPIA database. d) Expression of EFNA1 in ESCC cancer and adjacent normal tissues tested by IHC staining (magnification 200x). e) Expression levels of EFNA1 in ESCC tumor and normal tissues were determined using western blotting analysis. f) Forest plot of relationship between overall survival and different characteristics (Age, gender, TNM stage, and EFNA1 expression). g) Statistical analyses of association between EFNA1 expression and overall survival in 76 patients with ESCC. **** p < 0.0001, compared with no metastasis group, were regarded as statistically acceptable.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: Relationship between EFNA1 expression and clinicopathological features of ESCC.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: Expressing

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: EFNA1 promotes ESCC cell proliferation, migration, and invasion. a) Relatively protein expression of EFNA1 by western blotting analysis in stable EFNA1‑knockdown KYSE‐150 cell line. b) Effect of EFNA1 on proliferation of shEFNA1‐transfected KYSE‐150 cells analyzed by colony‐formation assay. c) Proliferative ability of KYSE‐150 cells transfected with shEFNA1 evaluated using RTCA assay. d) Migration of shEFNA1‐transfected KYSE‐150 cells after transfection by wound‐healing assay. e, f) Effect of EFNA1 on migration and invasion of KYSE‐150 cells transfected with shEFNA1 tested by Transwell assays, penetrated cells were counted after 48 h of incubation and analyzed by Image J. g, h) Knockdown of EFNA1 inhibits the tumor growth, in vivo tumorigenicity of stable EFNA1‑knockdown KYSE‐150 cell line in xenograft subcutaneous tumor and lung metastases model were tested by live imaging. i) HE staining of lung metastases. * p < 0.05, ** p < 0.01, **** p < 0.0001, compared with shNC control group, were regarded as statistically acceptable.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: Migration, Expressing, Western Blot, Transfection, Colony Assay, Wound Healing Assay, Incubation, Knockdown, In Vivo, Imaging, Staining, Control

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: ESCC‐specific aptamer screening, affinity, specificity, and tissue targeting analysis. a) High enrichment of selection library, selected aptamers were named EA1, EA2, EA3, EA4, and EA5. b) Flow cytometry assays of FAM‐labeled aptamer candidates to target KYSE‐150 cells. c) Secondary structures of EA1 and EA2 predicted by RNAfold. d) LSCM imaging of target KYSE‐150 and control HEEC cells incubated with 250 nM FAM‐labeled EA1 and EA2. e) Determination of dissociation constant (K d ) of EA1 and EA2 to KYSE‐150 cells using flow cytometry. f) Binding assays of FAM‐labeled aptamer EA1 (blue line) and EA2 (yellow line) to different cell lines by flow cytometry. FAM‐labeled random ssDNA (red line) was used as control. g) Confocal imaging of paired ESCC clinical tissues with FAM‐labeled EA1 and EA2 compared with paired adjacent tissues. h) In vivo targeting ability and biodistribution of the aptamer EA1 in xenograft subcutaneous tumor model. i) In vivo targeting ability and biodistribution of aptamer EA1 in xenograft lung metastases model.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: Selection, Flow Cytometry, Labeling, Imaging, Control, Incubation, Binding Assay, In Vivo

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: In vitro ESCC cancer suppression efficiency of EA1‐modified PTX/siEFNA1‐loaded nanomedicine. a, b) Expression of EFNA1 in KYSE‐150 cells after treating with various NDDS. c) Proliferative ability of KYSE‐150 cells after treating with various NDDS evaluated using RTCA assay. d) The representative scattering plots of apoptotic assay of KYSE‐150 cells treated with free drugs or various NDDS using Annexin V‐FITC apoptosis kit. e, f) Inhibition effects of various NDDS on migration and invasion of KYSE‐150 cells tested by Transwell assays, penetrated cells were counted after 48 h of incubation and analyzed by Image J. Treatments includes PBS (#1), EYLNs (#2), EA1‐EYLNs (#3), EYLNs‐siEFNA1 (#4), EA1‐EYLNs‐siEFNA1 (#5), PTX (#6), EYLNs‐PTX (#7), EA1‐EYLNs‐PTX (#8), EYLNs‐PTX/siEFNA1 (#9), and EA1‐EYLNs‐PTX/siEFNA1 (#10). ** p < 0.01, *** p < 0.001, **** p < 0.0001, compared with PBS group, were regarded as statistically acceptable.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: In Vitro, Modification, Expressing, Inhibition, Migration, Incubation

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: In vivo ESCC cancer suppression efficiency of EA1‐EYLNs‐PTX/siEFNA1 in KYSE‐150 tumor‐bearing mice. a) Hemolysis test of EA1‐EYLNs‐PTX/siEFNA1 at different concentrations. b) Schematic diagram of therapeutic treatment protocol in luciferase‐expressing KYSE‐150 tumor‐bearing mice. Treatment includes PBS (#1), EYLNs (5 nmol, #2), EYLNs‐EA1 (5 nmol, #3), EYLNs‐siEFNA1 (5 nmol, #4), EA1‐EYLNs‐siEFNA1 (5 nmol, #5), PTX (120 nmol, #6), EYLNs‐PTX (5 nmol EYLNs with 120 nmol PTX, #7), EA1‐EYLNs‐PTX (#8), EYLNs‐PTX/siEFNA1 (#9), and EA1‐EYLNs‐PTX/siEFNA1 (#10). c, d) Luciferase signals of ESCC tumors in luciferase‐expressing KYSE‐150 tumor‐bearing mice. Luciferase signals were quantified every week for 4 weeks. e) Relative tumor volume changes measured and calculated every 3 d of luciferase‐expressing KYSE‐150 tumor‐bearing mice under different treatments. f, g) Tumor images and tumor weights of isolated tumor tissues in different NDDS‐treated groups.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: In Vivo, Luciferase, Expressing, Isolation

Journal: Advanced Science

Article Title: Development of a Specific Aptamer‐Modified Nano‐System to Treat Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202309084

Figure Lengend Snippet: In vivo ESCC cancer suppression efficiency of EA‐EYLNs‐PTX/siEFNA1 in KYSE‐150 derived lung metastasis mice. a) Schematic diagram of therapeutic treatment protocol in BALB/c nude mice with KYSE‐150 derived lung metastasis. Treatments includes PBS, EA1‐EYLNs‐siEFNA1, EA1‐EYLNs‐PTX, and EA1‐EYLNs‐PTX/siEFNA1. b–d) Luciferase signals of lung metastasis in vivo (b) and isolated lung (c‐d) after 5 times of treatments. e) HE staining of lung metastasis sections. Scale bar: 5000 mm and 0.2 mm.

Article Snippet: The human esophageal squamous cell carcinoma KYSE‐150 was maintained in the laboratory and authenticated by the China Center for Type Culture Collection (CCTCC).

Techniques: In Vivo, Derivative Assay, Luciferase, Isolation, Staining

Journal: Molecular Therapy. Nucleic Acids

Article Title: SOX2-Upregulated microRNA-30e Promotes the Progression of Esophageal Cancer via Regulation of the USP4/SMAD4/CK2 Axis

doi: 10.1016/j.omtn.2020.10.027

Figure Lengend Snippet: SOX2 Is Highly Expressed in EC Tissues and Cells, which Is Negatively Correlated with EC Patients’ Prognosis (A) mRNA expression of SOX2 in EC tissues and adjacent normal tissues detected by qRT-PCR (n = 30). (B) Quantitation of SOX2 protein in EC tissues and adjacent normal tissues by immunohistochemical assay. (C) Kaplan-Meier analysis showing the correlation of SOX2 expression with OS and DFS of EC patients. (D) SOX2 expression determined by qRT-PCR and western blot analysis in EC cell lines (Eca109, EC9706, KYSE150, and TE-1) and normal esophageal epithelial cells (HEEC). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Paired t test was used for comparison between data of EC tissues and adjacent normal tissues, and one-way ANOVA was used for data comparison among multiple groups. Data are shown as mean ± standard deviation of three technical replicates.

Article Snippet: Human esophageal epithelial cell line HEEC and human esophageal cancer cell lines (Eca109, EC9706, KYSE150, and TE-1) were attained from the Cell Bank, China Center for Type Culture Collection (Wuhan, Hubei, P.R.

Techniques: Expressing, Quantitative RT-PCR, Quantitation Assay, Immunohistochemical staining, Western Blot, Comparison, Standard Deviation

Journal: Molecular Therapy. Nucleic Acids

Article Title: SOX2-Upregulated microRNA-30e Promotes the Progression of Esophageal Cancer via Regulation of the USP4/SMAD4/CK2 Axis

doi: 10.1016/j.omtn.2020.10.027

Figure Lengend Snippet: SOX2 Promotes Proliferation, Migration, and Invasion, as well as EMT of Eca109 Cells via miR-30e Upregulation In Vitro (A) Expression of miR-30e in EC tissues and adjacent normal tissues (n = 30) determined by qRT-PCR. (B) Expression of miR-30e in two EC cell lines (Eca109 and TE-1) and normal esophageal epithelial cells detected by qRT-PCR. (C) Pearson correlation analysis of SOX2 expression with miR-30e expression. (D) Expression of SOX2 and miR-30e in Eca109 cells transfected with oe-SOX2 detected by qRT-PCR. Eca109 cells were transfected with oe-SOX2 or in combination with miR-30e inhibitor. (E) Expression of SOX2 and miR-30e in Eca109 cells measured by qRT-PCR. (F) Proliferation of Eca109 cells detected by EdU assay (×200). (G) Quantitation of migration of Eca109 cells detected by scratch test. (H) Quantitation of invasion of Eca109 cells detected by Transwell assay. (I) Quantitative western blot analysis of EMT-related proteins (E-cadherin, N-cadherin, and Vimentin) in Eca109 cells. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Paired t test was used for comparison between data of EC tissues and adjacent normal tissues, while unpaired data between the other two groups were compared by unpaired t test. Comparisons among multiple groups were performed using one-way ANOVA. Data are shown as mean ± standard deviation of three technical replicates.

Article Snippet: Human esophageal epithelial cell line HEEC and human esophageal cancer cell lines (Eca109, EC9706, KYSE150, and TE-1) were attained from the Cell Bank, China Center for Type Culture Collection (Wuhan, Hubei, P.R.

Techniques: Migration, In Vitro, Expressing, Quantitative RT-PCR, Transfection, EdU Assay, Quantitation Assay, Transwell Assay, Western Blot, Comparison, Standard Deviation