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SUMMARY: Esophageal cancer is one of the most aggressive gastrointestinal cancers. Invasion and metastasis are the main causes of poor prognosis of esophageal cancer. SPRY2 has been reported to exert promoting effects in human cancers, which controls signal pathways including PI3K/AKT and MAPKs. However, the expression of SPRY2 in esophageal squamous cell carcinoma (ESCC) and its underlying mechanism remain unclear. In the present study, we aimed to investigate the detailed role of SPRY2 in the regulation of cell proliferation, invasion and ERK/AKT signaling pathway in ESCC. It was identified that the expression level of SPRY2 in ESCC was remarkably decreased compared with normal tissues, and it was related to clinicopathologic features and prognosis ESCC patients. The upregulation of SPRY2 expression notably inhibited the proliferation, migration and invasion of Eca-109 cells. In addition, the activity of ERK /AKT signaling was also suppressed by the SPRY2 upregulation in Eca-109 cells. Our study suggests that overexpression of SPRY2 suppress cancer cell proliferation and invasion of by through suppression of the ERK/AKT signaling pathways in ESCC. Therefore, SPRY2 may be a promising prognostic marker and therapeutic target for ESCC.
El cáncer de esófago es uno de los cánceres gastrointestinales más agresivos. La invasión y la metástasis son las principales causas de mal pronóstico del cáncer de esófago. Se ha informado que SPRY2 ejerce efectos promotores en los cánceres humanos, que controla las vías de señales, incluidas PI3K/AKT y MAPK. Sin embargo, la expresión de SPRY2 en el carcinoma de células escamosas de esófago (ESCC) y su mecanismo subyacente aún no están claros. En el presente estudio, nuestro objetivo fue investigar el papel detallado de SPRY2 en la regulación de la proliferación celular, la invasión y la vía de señalización ERK/AKT en ESCC. Se identificó que el nivel de expresión de SPRY2 en ESCC estaba notablemente disminuido en comparación con los tejidos normales, y estaba relacionado con las características clínico-patológicas y el pronóstico de los pacientes con ESCC. La regulación positiva de la expresión de SPRY2 inhibió notablemente la proliferación, migración e invasión de células Eca-109. Además, la actividad de la señalización de ERK/AKT también fue suprimida por la regulación positiva de SPRY2 en las células Eca-109. Nuestro estudio sugiere que la sobreexpresión de SPRY2 suprime la proliferación y la invasión de células cancerosas mediante la supresión de las vías de señalización ERK/AKT en ESCC. Por lo tanto, SPRY2 puede ser un marcador de pronóstico prometedor y un objetivo terapéutico para la ESCC.
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Humans , Esophageal Neoplasms/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , Esophageal Squamous Cell Carcinoma/metabolism , Membrane Proteins/metabolism , Immunohistochemistry , Biomarkers, Tumor , Blotting, Western , Extracellular Signal-Regulated MAP Kinases , Cell Proliferation , Proto-Oncogene Proteins c-aktABSTRACT
@#Objective To investigate the effects of exosome-derived miR-1246 on metastasis and autophagy of esophageal squamous cell carcinoma(ESCC).Methods The exosomes of ESCC cells were extracted by exosome extraction kit,the morphology was observed under electron microscope,the particle size was detected by nano-particle size analyzer,and the expression of exosome markers TSG101 and Calnexin was detected by Western blot.The RNA of exosomes of ESCC cells and normal esophageal epithelial cells were sequenced to analyze the differentially expressed miRNAs,which were analyzed by KEGG Pathway enrichment.The expression of miR-1246 in tumor tissues and paracancer tissues of 155 patients with ESCC was evaluated by transcriptome sequencing.The effect of miR-1246 on the migration ability of ESCC cells was detected by Transwell assay and the effect on autophagy was detected by Western blot.Results The exosomes derived from ESCC cells were intact in shape and similar to spherical vesicle-like structure with the particle size of 50～80 nm.The exosome marker TSG101 protein was positive and Calnexin protein was negative.There were 59 common differentially expressed miRNAs between exosomes of ESCC cells and normal esophageal epithelial cells.Exosome miR-1246 was highly expressed in ESCC cells,and the differentially expressed miRNAs were mainly enriched in autophagy metabolic pathway.Exosome miR-1246 was highly expressed in cancer tissues of patients with ESCC.Overexpression of miR-1246 significantly promoted the migration and autophagy of ESCC cells(t=4.119 and 48.150,P <0.05 and <0.001,respectively).Inhibition of miR-1246 expression inhibited the migration and autophagy of ESCC cells(t=9.067 and 51.270,P <0.01 and <0.001,respectively).Conclusion miR-1246 derived from exosomes can significantly affect the metastasis and autophagy of ESCC cells.
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@#ObjectiveTo detect the gene variation and expression of PLCH1 in esophageal squamous cell carcinoma(ESCC),analyze the function of PLCH1 gene in ESCC and explore its mechanism.MethodsThe copy number variation of PLCH1 in ESCC was analyzed by GISTIC,and the expression of PLCH1 in ESCC and normal esophageal tissues was detected by TCGA database and immunohistochemistry method. The expression of PLCH1 in ESCC cell lines was detected by real-time fluorescence quantitative PCR(qPCR) and Western blot,and the effects of PLCH1 silencing on the proliferation and migration of ESCC cells were detected by MTT assay,colony formation assay and Transwell assay.Results There was significant copy number amplification of PLCH1 in ESCC(G-scores > 0. 1,P < 0. 05),and the expression levels of PLCH1 mRNA and protein in ESCC were significantly higher than those in normal tissues(F = 36. 00 ～ 1 101. 00respectively,each P < 0. 000 1). After PLCH1 silencing,the ability of proliferation,clone formation and migration of ESCC cells KYESE180 and TE-9 decreased significantly(F = 35. 49 ～ 634. 00 respectively,each P < 0. 001).Conclusion PLCH1 plays an oncogenic role in ESCC,which is of great significance for the metastasis and proliferation of ESCC,and can be used as a potential target for the treatment of ESCC.
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@#Objective To study the expression of FAM84B in esophageal squamous cell carcinoma(ESCC) and its regulatory mechanism on cell growth by p53 pathway.Methods A total of 508 ESCC tumor samples and their adjacent normal tissue samples were collected from Shanxi Cancer Hospital and Affiliated Cancer Hospital of Xinjiang Medical University,which are high incidence areas of ESCC in China.Using whole genome sequencing(WGS) and whole exome sequencing(WES),FAM84B gene with significantly amplified copy number were screened.In ESCC cell line KYSE150with high expression of FAM84B,the effect of FAM84B on cell growth was detected by MTT and hard clone formation assay after interfering with FAM84B by small interfering RNA infection;In ESCC cell line KYSE450 with low expression of FAM84B,FAM84B was overexpressed with plasmid pCMV-3 ×flag-FAM84B,and the effect of FAM84B on cell growth was detected by MTT and hard clone formation assay.The effects of FAM84B knock-down on the expression of CDK4,CDK6and CCND1 were detected by Western blot.Results WGS analysis of 508 ESCC paraffin sections showed 109 cases of FAM84B copy number amplification(21.45%);FAM84B gene existed near 8q24.21 and was a significantly enlarged lesion peak.Knockdown of FAM84B inhibited the proliferation of ESCC cells,while overexpression of FAM84B promoted that.The low expression of FAM84B promoted the cell cycle progression mediated by p53.Conclusion FAM84B can promote cell proliferation by promoting the cell cycle transition from G1 phase to S phase via inhibiting the expression of p53 pathway proteins in ESCC cells.
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Objective:To analyze the local recurrence patterns after concurrent chemoradiotherapy (CCRT) for thoracic esophageal squamous cell carcinoma (ESCC) through image fusion, and to explore the risk factors of local recurrence and its relationships with dosimetric indices.Methods:A retrospective analysis was conducted for 209 thoracic ESCC patients who received radical CCRT in Fourth Hospital of Hebei Medical University during 2016-2019. For the patients diagnosed as the local recurrence of esophageal lesions, their CT images were fused with the original planning CT images using image registration software to identify the recurrence sites. Through 1∶1 propensity score matching (PSM) of the clinal data of patients with local recurrence (the recurrence group, nbefore = 81, nafter = 62) and those without local recurrence (the recurrence-free group, nbefore = 128, nafter=62), the dose and volume parameters of the treatment plans for the two groups were compared. Univariate and multivariate analyses were conducted using the Kaplan-Meier method and the Cox regression model to analyze the factors affecting the overall survival (OS), progression-free survival (PFS), and recurrence-free survival (RFS). Results:All patients had 1-, 3-, and 5-year OS rates of 80.9%, 42.6%, and 33.0%, respectively, 1-, 3-, and 5-year PFS rates of 67.9%, 34.0%, and 27.9%, respectively, and 1-, 3-, and 5-year RFS rates of 71.3%, 39.2%, and 30.5%, respectively. T stage, N stage, and radiation dose were independent prognostic factors for the OS, PFS, and RFS ( HR = 1.42-1.87, P < 0.05) of the patients, respectively. Among 68 patients with local recurrence, 62 cases (91.2%) suffered recurrence within the gross tumor volume (GTV). The dose and volume parameters of patients with local recurrence, such as GTV- D95%, clinical target volume (CTV)- D95%, GTV- D50%, CTV- D50%, and planning target volume (PTV)- D50%, GTV- V60, CTV- V60, and PTV- V60, were significantly lower than those of patients free from the local recurrence ( t=1.90-2.15, P < 0.05). Conclusions:Local recurrence of patients with thoracic ESCC after radical CCRT occurs mainly within the GTV. Increasing radiation doses may contribute to their survival benefits. The D50% for each target volume in the radiotherapy plan may be related to local recurrence, and it is necessary to conduct further research.
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OBJECTIVE: The long non-coding RNA (lncRNA) KCNQ1 overlapping transcript 1 (KCNQ1OT1) exerts vital regulatory functions in diverse tumors. However, the biological function of KCNQ1OT1 in esophageal squamous cell carcinoma (ESCC) remains unclear. METHODS: KCNQ1OT1 expression was detected in ESCC tissues using quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation, apoptosis, migration, and invasion were detected by the CCK-8 assay, EdU assay, flow cytometry analysis, and Transwell experiments, respectively. Bioinformatics analysis, luciferase reporter experiments, and RNA immunoprecipitation assays were used to predict and validate the regulatory relationships between KCNQ1OT1, microRNA-133b (miR-133b) and epidermal growth factor receptor (EGFR). RESULTS: KCNQ1OT1 expression was remarkably upregulated in ESCC tissues and cell lines. Overexpression of KCNQ1OT1 markedly promoted ESCC cell proliferation, migration, and invasion and enhanced the expression of N-cadherin, MMP-2, and MMP-9, but inhibited apoptosis and E-cadherin expression in ESCC cell lines; KCNQ1OT1 knockdown exerted the opposite effects. KCNQ1OT1 could directly bind to miR-133b and suppress its expression, and miR-133b reversed the effects of KCNQ1OT1 overexpression in ESCC cells. MiR-133b reduced the expression of epidermal growth factor receptor (EGFR); further, KCNQ1OT1 activated the phosphatidylinositol 3-kinase/AKT serine/threonine kinase 1 (PI3K/AKT) signaling pathway by repressing miR-133b repression and indirectly upregulating EGFR. KCNQ1OT1 expression was positively correlated with EGFR mRNA expression and negatively correlated with miR-133b expression. CONCLUSION: KCNQ1OT1 facilitates ESCC progression by sponging miR-133b and activating the EGFR/PI3K/AKT pathway.
Subject(s)
Humans , Esophageal Neoplasms/genetics , MicroRNAs/genetics , RNA, Long Noncoding/genetics , Esophageal Squamous Cell Carcinoma/genetics , Phosphatidylinositol 3-Kinases , Cell Proliferation/genetics , KCNQ1 Potassium Channel/geneticsABSTRACT
The aim of this study was to develop a diagnostic strategy for esophageal squamous cell carcinoma(ESCC) that combines plasma metabolomics with machine learning algorithms.Plasma-based untargeted metabolomics analysis was performed with samples derived from 88 ESCC patients and 52 healthy controls.The dataset was split into a training set and a test set.After identification of differential me-tabolites in training set,single-metabolite-based receiver operating characteristic (ROC) curves and multiple-metabolite-based machine learning models were used to distinguish between ESCC patients and healthy controls.Kaplan-Meier survival analysis and Cox proportional hazards regression analysis were performed to investigate the prognostic significance of the plasma metabolites.Finally,twelve differential plasma metabolites (six up-regulated and six down-regulated) were annotated.The pre-dictive performance of the six most prevalent diagnostic metabolites through the diagnostic models in the test set were as follows:arachidonic acid (accuracy:0.887),sebacic acid (accuracy:0.867),indoxyl sulfate (accuracy:0.850),phosphatidylcholine (PC) (14:0/0:0) (accuracy:0.825),deoxycholic acid(accuracy:0.773),and trimethylamine N-oxide (accuracy:0.653).The prediction accuracies of the ma-chine learning models in the test set were partial least-square (accuracy:0.947),random forest (accu-racy:0.947),gradient boosting machine (accuracy:0.960),and support vector machine (accuracy:0.980).Additionally,survival analysis demonstrated that acetoacetic acid was an unfavorable prognostic factor(hazard ratio (HR):1.752),while PC (14:0/0:0) (HR:0.577) was a favorable prognostic factor for ESCC.This study devised an innovative strategy for ESCC diagnosis by combining plasma metabolomics with machine learning algorithms and revealed its potential to become a novel screening test for ESCC.
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Objective: To investigate the expression of long non-coding RNA(lncRNA) SNHG3 in esophageal squamous cell carcinoma (ESCC) and its effect on the migration and invasion of ECA-109 cells. Methods: Samples of tumor and corresponding para-carcinoma tissues were collected from 60 patients with ESCC, who were enrolled in Anyang Tumor Hospital from June 2011 to June 2014. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of lncRNA SNHG3 in ESCC tumor and para-carcinoma tissues, ECA-109 ESCC cells, and HEEC human normal esophageal epithelial cells. The correlation between lncRNA SNHG3 expression and clinical characteristics of patients with ESCC was assessed. ECA-109 cells were transfected with siRNA-SNHG3 plasmids to knockdown lncRNA SNHG3 expression. The correlation between lncRNA SNHG3 expression and patient prognosis was determined by Kaplan-Meier analysis. Transwell assay was used to investigate the migration and invasion abilities of ECA-109 cells. SNAIL and TWIST mRNA and protein levels in ECA-109 cells were detected by qRT-PCR and Western blot assay, respectively. Results: The expression level of lncRNA SNHG3 was significantly higher in ESCC tumor tissues compared to that in pericarcinomatous tissues and in ECA-109 cells compared to that in HEEC cells (both P<0.05). LncRNA SNHG3 expression in ESCC cells was significantly correlated with tumor differentiation, TNM stage, and lymph node metastasis (P< 0.05). LncRNA SNHG3 expression in ECA-109 cells decreased significantly upon transfection with the siRNA-SNHG3 plasmid (P<0.05). High expression of lncRNA SNHG3 was significantly correlated with poor prognosis in patients with ESCC. After lncRNA SNHG3 knockdown, the migration and invasion of ECA-109 cells and the mRNA and protein expression levels of SNAIL and TWIST were reduced significantly (P<0.05). Conclusions: LncRNA SNHG3 is highly expressed in ESCC tumor tissues and cells and promotes the migration and invasion of ECA-109 cells by regulating the expression of SNAIL and TWIST proteins.
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@#[Abstract] Objective: To investigate the effect of long non-coding RNA DiGeorge syndrome critical region gene 5 (DGCR5) on proliferation, invasion and migration of esophageal squamous cell carcinoma (ESCC) TE1 cells and its mechanism. Methods: qPCR was used to detect the expression level of DGCR5 in ESCC cell lines (TE1, Yes-2, KYSE150 and Eca9706). TE1 cells were transfected with siRNA-DGCR5(si-DGCR5) and negative control (si-NC) plasmids, respectively. CCK-8, Wound healing and Transwell assay were used to detect the proliferation, migration and invasion of TE1 cells before and after DGCR5 knockdown. The relationship between DGCR5 expression and epidermal growth factor receptor (EGFR) in ESCC tissues was analyzed by GEPIA database. The mRNA and protein expressions of EGFR in ESCC cell line were examined by qPCR and Western blotting (WB). WB was further used to detect the expression of EGFR protein in TE1 cells before and after DGCR5 knockdown. Results: lncRNA DGCR5 was highly expressed in ESCC cell lines (all P<0.01). qPCR confirmed that the expression of DGCR5 in TE1 cells of si-DGCR5 group was significantly lower than that of si-NC group (P<0.01). The proliferation, migration and invasion ability of TE1 cells in si-DGCR5 group were significantly lower than those in si-NC group (all P<0.01). GEPIA database showed that the expression of DGCR5 was positively correlated with EGFR in ESCC tissues (P<0.01). WB showed that the protein level of EGFR in TE1 cells of si-DGCR5 group decreased significantly (P<0.01). Conclusion: lncRNA DGCR5 is highly expressed in ESCC cells, and promotes the proliferation, invasion and migration of TE1 cells possibly by up-regulating EGFR expression.
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@#[Abstract] Objective: To investigate the expression of long non-coding RNA (lncRNA) DiGeorge syndrome critical region gene 5 (DGCR5) in esophageal squamous cell carcinoma (ESCC) tissues, and to analyze its relationship with clinicopathological features and prognosis of ESCC patients. Methods: The expression of DGCR5 in ESCC data set from TCGA database was analyzed by bioinformatics method. Sixty pairs of ESCC tissues and para-cancerous tissues resected at the Fourth Hospital of Hebei Medical University from August 2016 to March 2017 were collected for this study. The expression of DGCR5 in ESCC tissues was detected by qPCR. The correlation between the expression of DGCR5 and the clinicopathological features and prognosis of ESCC patients was analyzed. Results: TCGAdatabase analysis showed that the expression of DGCR5 in ESCC tissues was significantly higher than that in normal esophageal tissues (P<0.01). The expression of DGCR5 in ESCC tissues was significantly higher than that in para-cancerous tissues (P<0.01). The expression level of DGCR5 was significantly correlated with TNM staging and lymph node metastasis in ESCC patients (all P<0.05). Kaplan-Meier univariate analysis showed that the 2-year survival rate of ESCC patients with high DGCR5 expression was significantly lower than that of patients with low expression (P<0.05). Conclusion: DGCR5 is highly expressed in ESCC tissues and is closely related to TNM staging, lymph node metastasis and poor prognosis, which may serve as a molecular marker for early diagnosis and prognosis prediction of ESCC.
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@#[Abstract] Objective: To investigate the changes in malignant biological behaviors and expression of programmed cell death-ligand 1 (PD-L1) in esophageal squamous cell carcinoma (ESCC) YES-2 cell line after cis-dichlorodiammine platinum (CDDP) induction (YES-2/CDDP-R). Methods: YES-2 cells were treated with CDDP from low concentration to high concentration (0.25-2.0 μg/ml) with intermittent impact (15-25 days per concentration) to establish ESCC CDDP-resistant cell line YES-2/CDDP-R. The morphological change of YES-2/CDDP-R cells was observed under the inverted microscope. Methyl thiazolyl tetrazolium (MTT) was used to detect cell sensitivity to CDDP. Wound healing assay was used to detect cell migration ability. qPCR and Western blotting were used to detect mRNA and protein expressions of PD-L1. Results: After CDDP gradien ttreatment for9 months,YES-2/CDDP-R cells were successfully established. The morphology of the YES-2/CDDP-R cells showed uneven size, intracellular vacuoles and significantly increased black particles along with the appearance of huge cells. The IC50 of CDDP for YES-2/CDDP-R cells was significantly higher than that for parental cells, indicating decreased sensitivity to CDDP (P<0.05). Compared to theYES-2 cells, the proliferation and migration of YES-2/CDDP-R cells were significantly increased (P<0.05 or P<0.01), and the mRNA and protein expressions of PD-L1 were significantly up-regulated (all P<0.001). Conclusion: YES-2 cells with CDDP resistance (YES-2/CDDP-R) were successfully established. The sensitivity of YES-2/CDDP-R cells to CDDP was significantly reduced while the abilities of cell proliferation and migration were enhanced. The up-regulation of PD-L1 in YES-2/CDDP-R cells suggests that CDDP-resistance could promote immune escape by inducing PD-L1 up-regulation.
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@#[Abstract] Objective: To investigate the effects and mechanisms of long non-coding RNA (lncRNA) non-coding RNA-activated by DNA damage (NORAD) on the proliferation and migration of esophageal squamous cell carcinoma (ESCC) EC9706 cells. Methods: RT-PCR was used to detect the mRNA expression level of NORAD in different ESCC cells (EC9706, TE1, YES-2, KYSE150). Small interfering RNA (siRNA) targeting NORAD gene was transfected into EC9706 cells (as si-NORAD group) with RNA interference technique to knockdown NORAD expression; in addition, blank control group (as Ctrl group, without any transfection) as well as negative control group (as NC group, transfected with siRNAnegative control sequence)werealsoestablished. qPCR was used to verify the transfection efficiency. MTT, Colony formation assay and Wound-healing test were used to detect the abilities of proliferation and migration of EC9706 cells before and after NORAD knockdown. Western blotting was used to detect the expressions of E-cadherin, N-cadherin and Snail in EC9706 cells before and after NORAD knockdown. Results: NORAD mRNAwas highly expressed in 4 ESCC cell lines. Comparing with TE1, YES-2 and KYSE150 cells, the expression of NORAD mRNA was significantly higher in EC9706 cells (P<0.01). After transfection of NORAD-siRNA into EC9706 cells, the expression of NORAD was down-regulated significantly as comparing with Ctrl group and NC group (all P<0.01), in the meanwhile, the proliferation and migration abilities of EC9706 cells were also significantly suppressed (P<0.05).After NORAD knockdown, the expression of E-cadherin was up-regulated while the expressions of N-cadherin and Snail were down-regulated in EC9706 cells (all P<0.05). Conclusion: NORAD is highly expressed in EC9706 cells;knockdown of NORAD expression can inhibit the proliferation and migration ability of EC9706 probably through up-regulating E-cadherin and down-regulating N-cadherin and Snail.
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@#[Abstract] Objective: To explore the mechanism of miR-145-5p on malignant biological behaviors, such as pro-liferation, invasion, migration and epithelial-mesenchymal transition (EMT), of esophageal squamous cell carcinoma (ESCC) TE-10 cells. Methods: The expression of miR-145-5p in ESCC cell lines and normal cells was detected by PCR. Dual luciferase reporter gene assay was used to detect the targeted regulation between miR-145-5p and insulin-like growth factor 1 receptor (IGF1R). The expres-sions of IGF1R protein and EMT related proteins were detected by Western blotting. Transwell assay and CCK-8 assay were carried out to detect the effects of miR-145-5p/IGF1R axis on the proliferation, migration andinvasionofTE-10 cells. Results: miR-145-5p was down-regulated in ESCC cell lines with the lowest expression in TE-10 cells (P<0.01orP<0.05).Over-expression of miR-145-5p significantly inhibited proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Dual luciferase reporter gene assay con-firmed that miR-145-5p targetedly down-regulated IGF1R expression (P<0.01). The restora-tion experiments further confirmed that simultaneous over-expression of miR-145-5p and IGF1R significantly attenuated the promotion effect of IGF1R on proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Conclusions: Over-expression of miR-145-5p inhibits proliferation, invasion, migration and EMT of ESCC TE-10 cells by down-regulating IGF1R.
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@#[Abstract] Objective: This study aimed at investigating the epigenetic regulation mechanism of abnormally low expression of SHP-1 gene in esophageal squamous cell carcinoma (ESCC). Methods:Atotal of 71 cases of ESCC tissues and corresponding para-cancer tissues(2 cm from the edge of the cancer) resected during surgery at the Department of thoracic surgery of Hebei Province, the Fourth Hospital of Hebei Medical University from 2008 to 2011 were collected for this study. The expression level of SHP-1 mRNA and protein was detected in esophageal cancer cell lines (Eca109, Kyse170, Yes-2) before and after 5-Aza-dC or TSA treatment by RT-qPCR and Western blotting methods respectively. The methylation status of CpG sites in promoter region of SHP-1 was analyzed by bisulfite genome sequencing (BGS) in three esophageal cancer cell lines before and after 5-Aza-dC treatment. The methylation status of SHP-1 was studied by methylation-specific polymerase chain reaction (MSP) method in esophageal cancer cell lines, ESCC tissues and para-cancer tissues. The association between the SHP-1 promoter methylation status and clinic pathological parameters were analyzed in ESCC patients. Dual-luciferase reporter assay systems method was applied to detect the impacts of methylation status of CpG island in SHP-1 promoter region on gene transcription activity. For prognostic analysis of SHP-1 methylation, survival curves were constructed using the Kaplan-Meier method and the log-rank. Results: After treated with 5-Aza-dC, the expression level of SHP-1 mRNA and protein was significantly up-regulated in Eca109, Kyse170 and Yes-2 cells, meanwhile the methylation status of SHP-1 was decreased (P<0.05). The expression level of SHP-1 had no obviously change after treated with trichostatin A(TSA). The methylation frequency of promoter in ESCC tumor tissues was significantly higher than that in corresponding para-cancer tissues (P<0.05). When stratified for clinic pathologic characteristics, methylation frequency of SHP-1 was associated with TNM stage, pathological differentiation, and LN metastasis (P<0.05). The mRNAexpression level of SHP-1 in the ESCC tissues with SHP-1 methylation was significantly decreased compared to the ESCC tissues with unmethylation of SHP-1 (P<0.05). It was associated with methylation of promoter (P<0.05). The activity of fluorescein reporter vector in methylase treatment group was significantly lower than that in untreated group (P<0.05), indicating that SHP-1expression can be silenced by methylation of SHP-1 promoter. The result of Kaplan-Meier shown that SHP-1 promoter methylation was correlated with ESCC patients’poor survival. Conclusion: The transcriptional activity of SHP-1 can be inhibited with hypermethylated SHP-1 promoter region. The hypermethylated SHP-1 promoter induced the silencing of SHP-1. Therefore, SHP-1 gene may serve as one of prognostic methylation biomarkers for ESCC patients.
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Dysregulation of mTORC1/mTORC2 pathway is observed in many cancers and mTORC1 inhibitors have been used clinically in many tumor types; however, the mechanism of mTORC2 in tumorigenesis is still obscure. Here, we mainly explored the potential role of mTORC2 in esophageal squamous cell carcinoma (ESCC) and its effects on the sensitivity of cells to mTOR inhibitors. We demonstrated that RICTOR, the key factor of mTORC2, and p-AKT (Ser473) were excessively activated in ESCC and their overexpression is related to lymph node metastasis and the tumor-node-metastasis (TNM) phase of ESCC patients. Furthermore, we found that mTORC1/ mTORC2 inhibitor PP242 exhibited more efficacious anti-proliferative effect on ESCC cells than mTORC1 inhibitor RAD001 due to RAD001-triggered feedback activation of AKT signal. Another, we demonstrated that down-regulating expression of RICTOR in ECa109 and EC9706 cells inhibited proliferation and migration as well as induced cell cycle arrest and apoptosis. Noteworthy, knocking-down stably RICTOR significantly suppresses RAD001-induced feedback activation of AKT/PRAS40 signaling, and enhances inhibition efficacy of PP242 on the phosphorylation of AKT and PRAS40, thus potentiates the antitumor effect of RAD001 and PP242 both and . Our findings highlight that selective targeting mTORC2 could be a promising therapeutic strategy for future treatment of ESCC.
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@#Objective: To investigate the expression of lncRNA LINC00886 in human esophageal squamous cell carcinoma (ESCC) tissues and cell lines, and its effects on proliferation, migration and invasion of Eca109 cells. Methods: The cancer tissues and corresponding para-cancerous tissues of 69 ESCC patients were collected in the biological specimen bank of the Fourth Hospital of Hebei Medical University from June 2014 to December 2016; the ESCC cell lines Eca109, TE13, TE1, Kyse150, Yes-2 and Kyse170 were also collected. LINC00886 gene expression in ESCC tissues and cell lines was detected by qPCR. Eca109 cells were transfected with pIRES2-LINC00886 and pIRES2-NC, respectively, and the overexpression efficiency of LINC00886 gene in Eca109 cells was detected by qPCR; MTS, clone formation assay, wound-healing assay and Transwell invasion assay were respectively used to detect the effect of LINC00886 over-expression on proliferation, migration and invasion ability of Eca109 cells. Results: The expression of LINC00886 gene in ESCC tissues was significantly lower than that in para-cancerous tissues (P<0.01), and its expression level was associated with tumor TNM stage and lymph node metastasis (both P<0.05). The expression level of LINC00886 gene in ESCC cell lines was also lower than that of the control group (all P<0.01). Compared with control group, the expression level of LINC00886 gene was significantly higher in Eca109 cells transfected with pIRES2-LINC00886 (both P<0.05). Compared with the control group, LINC00886 overexpression significantly inhibited the proliferation, migration and invasion abilities of Eca109 cells (all P<0.01). Conclusion: The decreased expression of LINC00886 gene may be related to the occurrence and development of ESCC. Over-expression of LINC00886 gene inhibits the proliferation, migration and invasion abilities of ESCC cells.
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@# Objective: To investigate the mechanism of miR-503 modulates radio-resistance of esophageal squamous cell carcinoma (ESCC) by targeting excision-repair cross-complementing 1 (ERCC1). Methods: The expression level of miR-503 in radio-resistant ESCC tumor tissues and KYSE140 and KYSE140R cells was detected by qPCR. The miR-503 mimic, miR-503 inhibitor or si-ERCC1 was transfected into KYSE140 and KYSE140R cells.After radiation treatment, the colony formation assay and CCK-8 assay were used to detect the proliferation of KYSE140R cells. Flow cytometry was used to detect apoptosis of KYSE140R cells. WB was used to detect changes in protein expression of ERCC1. Dual luciferase reporter gene assay was used to validate the targeting relationship between miR-503 and ERCC1. Results: The expression level of miR-503 was down-regulated in radio-resistant tissues and ESCC cell lines (all P<0.01). Over-expression of miR-503 significantly inhibited cell proliferation and promoted apoptosis of KYSE140R cells (all P<0.01). Dual-luciferase reporter assay validated that ERCC1 was a target gene of miR-503, and miR-503 negatively regulated the expression of ERCC1. Over-expression of miR-503 significantly down-regulated the expression of ERCC1 in KYSE140 and KYSE140R cells (both P<0.01), inhibited cell proliferation (both P<0.01), but significantly increased apoptosis rate (all P<0.01); knockdown of ERCC1 exhibited a similar effect, while knockdown of both ERCC1 and miR-503 reversed the above effects. Conclusion: Over-expression of miR-503 up-regulated the radio-sensitivity of KYSE140R cells by targeting ERCC1.
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Esophageal cancer is a common malignant tumor of the upper gastrointestinal tract. Early symptoms of the disease are inconspicuous and the disease is often diagnosed at a later stage, leading to higher morbidity and mortality. Esophageal cancer morbidity and mortality in both genders ranks among the top 10 most common cancers. Early detection and early treatment are effective means to reduce the incidence and mortality of esophageal cancer. Tumor markers play an important role in early diagnosis, treatment monitoring and prognosis evaluation of esophageal cancer. This paper reviews the clinical application of tumor markers related to esophageal cancer and the exploration and application progress of new tumor markers for esophageal cancer.
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@#Objective: To investigate the relationship between PET/CT metabolic parameters and pathological features and prognosis in esophageal squamous cell carcinoma (ESCC) patients with intramural gastric metastasis (IGM). Methods: Totally 86 cases of ESCC IGM patients treated in Anhui Provincial Hospital Affiliated to Anhui Medical University from January 2008 to December 2014 were selected for this study. The patients received the imaging examination by positron emission tomography and computed tomography (PET/CT). The metabolic parameters including maximum standard uptake value (SUVmax), metabolic tumor volume (MTV), PET tumor length (PTL) and mean standard uptake value (SUVmean) were examined to calculate the total lesion glycolysis (TLG). The survival of the patients during 5-year follow-up was recorded, and the relationship between metabolic parameters and clinical pathological features and prognosis was analyzed. Results: SUVmax and SUVmean of IGM patients were related to the diameter of the primary tumor (all P<0.05); MTV was associated with the tumor diameter, lymph node metastasis, and TNM staging (all P<0.05); TLG was associated with the tumor diameter, lymph node metastasis, TNM stage, and tissue differentiation (all P<0.05). During the 5-year follow-up, 6 patients were lost to follow-up, 36 patients died and 44 patients survived; SUVmax, MTV, TLG, PTL, SUVmean, and TNM staging were predictors for patients’prognosis (all P<0.05); MTV, TLG, PTL, SUVmean, and TNM staging were risk factors for prognosis (all P< 0.05). Conclusion: The metabolic parameters including SUVmax, MTV, TLG, PTL and SUVmean in ESCC patients with IGM are related to the pathological characteristics of patients; moreover, MTV, TLG, PTL, SUVmean and TNM staging are risk factors for prognosis; so, PET/CT examination has certain clinical value for the prognosis assessment in ESCC patients with IGM.
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Objective To investigate the relationship between albumin/globulin ratio (AGR) and postoperative survival among patients with esophageal squamous cell carcinoma(ESCC) undergoing radical oesophagectomy, to establish an effective prognostic nomogram for ESCC and to provide a reference for prognosis prediction of ESCC. Methods From February 2014 to September 2017, 390 ESCC patients who underwent surgery were retrospectively enrolled from the tumor hospital in Fujian Province. The receiver operating characteristic curves(ROC) were applied to establish optimal cutoff points. Chi-square test was used to estimate the relationship between the AGR and the clinical features. Cox proportional hazards model was used to estimate the HR and 95% CI for the associations between AGR and Prognosis of ESCC patients. A nomogram model was established to predict the outcome of ESCC patients. Results The ROC demonstrated the best cutoff value for AGR was 1.16. A total of 356 patients were recruited in the final analysis, who were divided into the high AGR group (≥1.16) and the low AGR group (<1.16) by the best cutoff value. Both 1-year and 3-year survival rates in the high AGR group were higher than those detected in the low AGR group(all P<0.05). Multivariate analysis showed that the T stage, N stage, and AGR were independent prognostic factors of overall survival(all P<0.05). The HR of T stage was 1.87 (T3-T4 vs T1-T2, 95% CI: 1.04-3.35); The HR of N stage was1.89 (N+ vs N0, 95% CI: 1.13-3.17); The HR of AGR was 0.57(the high AGR group vs the low AGR group, 95%CI: 0.36-0.90). The concordance index(C-index) of the nomogram to predict overall survival was 0.68 (95% CI: 0.62-0.73，P<0. 001)．Conclusions The AGR was an independent prognosis factor for operated ESCC patients. The prognosis of ESCC in the high AGR group was better than that in the low AGR group and the prognostic nomogram provides individualized risk estimate of survival in ESCC patients after surgery.