Overall Survival Of Esophageal Squamous Cell Carcinoma Patients Research Articles

Novel miRNA markers and their mechanism of esophageal squamous cell carcinoma (ESCC) based on TCGA

MicroRNAs(miRNAs) are promising biomarkers for early esophageal squamous cell carcinoma (ESCC) detection and prognostic prediction. This study aimed to explore the potential biomarkers and molecular pathogenesis in the early diagnosis of ESCC. Firstly, 48 differentially expressed miRNAs (DEMs) and 1319 differentially expressed genes (DEGs) were identified between 94 ESCC tissues and 13 normal esophageal tissues in TCGA. From miRNA–mRNA regulatory network, there are 6558 target genes of the 48 DEMs, where 400 target genes are also among 1319 DEGs. Then, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment indicate that the 400 DEGs significantly enriched in cell cycle, proteoglycans in cancer, p53 signaling pathway, protein digestion and absorption, transcriptional dysregulation in cancer, and oocyte meiosis. And there are 66 DEGs among these six biological pathways, which we called GO-DEGs. From miRNA–mRNA regulatory network, 32 DEMs regulated the 66 GO-DEGs, where 22 DEMs were verified by different types of experiments in ESCC tissues, cells, or serum from the literature. For the other novel 10 DEMs, single-factor Cox regression analysis show that only hsa-miR-34b-3p showed no significant correlation with the overall survival of ESCC patients. Finally, we obtained the novel 9 ESCC-related DEMs, where three are down-regulated, and six are up-regulated. We analyzed the expression trends of target genes for five miRNAs and identified three significantly different miRNAs (hsa-miR-205-3p, hsa-miR-452-3p, and hsa-miR-6499-3p) confirmed by qPCR. Moreover, the stage-specific miRNAs were also suggested. These three qPCR validated miRNAs are also specific to the early stages of ESCC: hsa-miR-452-3p is specific to Stage I, II and III; hsa-miR-205-3p is specific in Stage II and III; and hsa-miR-6499-3p is Stage II specific. They might be the potential biomarkers for ESCC stage diagnosis. This study identified three novel miRNA markers potentially related to the diagnosis of ESCC and participated in the occurrence and development of ESCC through cell cycle, proteoglycans in cancer, p53 signaling pathway, protein digestion and absorption, transcriptional dysregulation in cancer, and signaling pathway for oocyte meiosis.

Suppressing SENP1 inhibits esophageal squamous carcinoma cell growth via SIRT6 SUMOylation.

Esophageal squamous cell carcinoma (ESCC) is a prevalent tumor in the gastrointestinal tract, but our understanding of the molecular mechanisms underlying ESCC remains incomplete. Existing studies indicate that SUMO specific peptidase 1 (SENP1) plays a crucial role in the development and progression of various malignant tumors through diverse molecular mechanisms. However, the functional mechanism and clinical implications of SENP1 in the progression of ESCC remain unclear. Bulk RNA-Sequencing (RNA-seq) was used to compare potential genes in the esophageal tissues of mice with ESCC to the control group. The up-regulated SENP1 was selected. The protein level of SENP1 in ESCC patient samples was analyzed by immunohistochemistry and western blot. The potential prognostic value of SENP1 on overall survival of ESCC patients was examined using tissue microarray analysis and the Kaplan-Meier method. The biological function was confirmed through in vitro and in vivo knockdown approaches of SENP1. The role of SENP1 in cell cycle progression and apoptosis of ESCC cells was analyzed by flow cytometry and western blot. The downstream signaling pathways regulated by SENP1 were investigated via using RNA-Seq. SENP1-associated proteins were identified through immunoprecipitation. Overexpression of Sirtuin 6 (SIRT6) wildtype and mutant was performed to investigate the regulatory role of SENP1 in ESCC progression in vitro. Our study discovered that SENP1 was upregulated in ESCC tissues and served as a novel prognostic factor. Moreover, SENP1 enhanced cell proliferation and migration of ESCC cell lines in vitro, as well as promoted tumor growth in vivo. Thymidine kinase 1 (TK1), Geminin (GMNN), cyclin dependent kinase 1(CDK1), and cyclin A2 (CCNA2) were identified as downstream genes of SENP1. Mechanistically, SENP1 deSUMOylated SIRT6 and subsequently inhibited SIRT6-mediated histone 3 lysine 56 (H3K56) deacetylation on those downstream genes. SIRT6 SUMOylation mutant (4KR) rescued the growth inhibition upon SENP1 depletion. SENP1 promotes the malignant progression of ESCC by inhibiting the deacetylase activity of SIRT6 pathway through deSUMOylation. Our findings suggest that SENP1 may serve as a valuable biomarker for prognosis and a target for therapeutic intervention in ESCC.

PET/CT deep learning prognosis for treatment decision support in esophageal squamous cell carcinoma

ObjectivesThe clinical decision-making regarding choosing surgery alone (SA) or surgery followed by postoperative adjuvant chemotherapy (SPOCT) in esophageal squamous cell carcinoma (ESCC) remains controversial. We aim to propose a pre-therapy PET/CT image-based deep learning approach to improve the survival benefit and clinical management of ESCC patients.MethodsThis retrospective multicenter study included 837 ESCC patients from three institutions. Prognostic biomarkers integrating six networks were developed to build an ESCC prognosis (ESCCPro) model and predict the survival probability of ESCC patients treated with SA and SPOCT. Patients who did not undergo surgical resection were in a control group. Overall survival (OS) was the primary end-point event. The expected improvement in survival prognosis with the application of ESCCPro to assign treatment protocols was estimated by comparing the survival of patients in each subgroup. Seven clinicians with varying experience evaluated how ESCCPro performed in assisting clinical decision-making.ResultsIn this retrospective multicenter study, patients receiving SA had a median OS 9.2 months longer than controls. No significant differences in survival were found between SA patients with predicted poor outcomes and the controls (p > 0.05). It was estimated that if ESCCPro was used to determine SA and SPOCT eligibility, the median OS in the ESCCPro-recommended SA group and SPOCT group would have been 15.3 months and 24.9 months longer, respectively. In addition, ESCCPro also significantly improved prognosis accuracy, certainty, and the efficiency of clinical experts.ConclusionESCCPro assistance improved the survival benefit of ESCC patients and the clinical decision-making among the two treatment approaches.Critical relevance statementThe ESCCPro model for treatment decision-making is promising to improve overall survival in ESCC patients undergoing surgical resection and patients undergoing surgery followed by postoperative adjuvant chemotherapy.Key PointsESCC is associated with a poor prognosis and unclear ideal treatments.ESCCPro predicts the survival of patients with ESCC and the expected benefit from SA.ESCCPro improves clinicians’ stratification of patients’ prognoses.Graphical

Molecular characteristics and multivariate survival analysis of 43 patients with locally advanced or metastatic esophageal squamous cell carcinoma.

Esophageal cancer (EC) is an aggressive malignant tumor with poor prognosis and high incidence. It is the sixth leading cause of cancer-related death in the world, and the 5-year overall survival (OS) rate is only 12-20%. The rapid development of next-generation sequencing (NGS) has provided powerful help for the treatment and management of EC patients. Tumor tissue and blood samples of 43 Chinese patients with nonsurgical esophageal squamous cell carcinoma (ESCC) were sequenced using a 425 gene-panel. Genomic profiling was explored and and the Cox proportional hazards model was used to analyze the correlations between gene or signaling pathway alterations and prognosis. In this study, the most common mutated genes were TP53 (90.5%), CCND1 (45.2%), FGF19 (38.1%), NOTCH1 (26.2%), PI3KCA (21.4%) and CDKN2A (19%). Among these mutations, PI3KCA and NOTCH1 showed mutual exclusion to some extent. In the univariate model, mutations in NOTCH1, CBLB and TSC2 genes and tumor mutation burden (TMB) ≥7 were independent biomarkers of OS. NOTCH1 (P=0.007, HR =2.87), CBLB (P=0.011, HR =4.68) and TSC2 (P=0.024, HR =3.7) were significantly associated with poorer OS, and patients with TMB ≥7 had longer OS (P=0.151, HR =0.31). In addition, patients who carried alteration in NOTCH signaling pathway had reduced OS (P=0.014, HR =2.54). NOTCH1, CBLB and TSC2 alterations were found to be potential indicators of poor prognosis in patients with ESCC. TMB was also positively correlated with the OS of ESCC patients, providing valuable insights for their treatment strategies.

Clinical and Pathologic Supraclavicular Lymph Node Metastases in Esophageal Squamous Cell Carcinoma Treated by Esophagectomy with Three-Field Lymph Node Dissection.

This study investigated the survival outcomes for surgically treated esophageal squamous cell carcinoma (ESCC) patients based on clinically suspicious supraclavicular lymph node (SCN) metastasis (cSCN+) and pathologically confirmed SCN metastasis (pSCN+). Using an institutional registry between 1994 and 2018, this study retrospectively analyzed 611 patients who received curative-intent esophagectomy with 3-field lymph node dissection for ESCC. The study used computed tomography and positron emission tomography to define cSCN+. Among 611 patients, 24.4% had cSCN+ and 12.2% had pSCN+. The 5-year overall survival (OS) rates were 68.2% for cN0, 43.5% for cN+ without cSCN+, and 30.3% for cN+ with cSCN+ (p = 0.018). Although the univariable analysis showed that cSCN+ was associated with poorer survival than cN0 or cN+ with cSCN- (hazard ratio [HR], 1.818; p < 0.001), the multivariable analysis did not support this finding (HR, 1.281; p = 0.681). The 5-year OS rates were 64.2% for pN0, 41.5% for pN+ without pSCN+, and 25.6% for pN+ with pSCN+ (p = 0.054). Univariable analysis showed an association of pSCN+ with poor OS (HR, 1.830; p < 0.001), but the difference in the multivariable analysis was not significant (HR, 0.912; p = 0.587). The presence of SCN metastasis did not have a significant impact on the OS of ESCC patients with 3-field lymph node dissectionregardless of clinical suspicion or pathologic confirmation.

A prognostic nomogram that includes MPV in esophageal squamous cell carcinoma.

Mean platelet volume (MPV), as a marker of platelet activity, has been shown to be an efficient prognostic biomarker in several types of cancer. Using MPV, this study aimed to create and validate a prognostic nomogram to the overall survival in esophageal squamous cell carcinoma (ESCC) patients. The nomogram was constructed and tested using data from a retrospective study of 1893 patients who were randomly assigned to the training and testing cohorts with a 7:3 randomization. In order to screen out the optimal predictors for overall survival (OS), we conducted the LASSO-cox regression, univariate, and multivariate cox regression analyses. Subsequently, the predictive accuracy of the nomogram was validated in both the training and the testing cohorts. Finally, decision curve analysis (DCA) was used to confirm clinical validity. Age, MPV, nerve invasion, T stage, and N stage were found as independent prognostic variables for OS and were further developed into a nomogram. The nomogram's prediction accuracy for 1-, 3-, and 5-year OS was 0.736, 0.749, 0.774, and 0.724, 0.719, 0.704 in the training and testing cohorts, respectively. Furthermore, DCA results indicated that nomograms outperformed the AJCC 8th and conventional T, N staging systems in both the training and testing cohorts. The nomogram, in conjunction with MPV and standard clinicopathological markers, could improve the accuracy of prediction of OS in ESCC patients.

Prognostic value of inflammatory markers and clinical features for survival in advanced or metastatic esophageal squamous cell carcinoma patients receiving anti-programmed death 1 treatment.

This study aims to assess the prognostic value of inflammatory markers and clinical features in advanced or metastatic esophageal squamous cell carcinoma (ESCC) patients receiving anti-programmed death 1 (PD-1) treatment. Based on receiver operating characteristic curve (ROC) analysis, Youden's indexes were applied to determine the cut-off values for inflammatory markers, including neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocye ratio (dNLR), monocyte-to-lymphocyte ratio (MLR), platelet-to-lymphocyte ratio (PLR), and systemic immune-inflammation index (SII). Wilcoxon test was conducted to evaluate the changes in above inflammatory markers. Kaplan-Meier method was utilized to estimate progression-free survival (PFS) and overall survival (OS), and the Log-rank test was used to compare the different survival between groups. Univariate and multivariate Cox regression analyses were performed to assess the prognostic value of inflammatory markers and clinical features. 162 advanced or metastatic ESCC patients receiving anti-PD-1 treatment were enrolled in this retrospective study. The cut-off values of NLR, dNLR, MLR, PLR, and SII were 4.748, 2.214, 0.309, 250.505, and 887.895, respectively. NLR, dNLR, PLR, and SII declined significantly among the partial response (PR) (P<0.001, P<0.001, P=0.036, P<0.001), objective response rate (ORR) (P<0.001, P<0.001, P=0.036, P<0.001), and disease control rate (DCR) (P<0.001, P<0.001, P=0.038, P<0.001) groups, respectively. Significant increases were found in NLR (P<0.001), dNLR (P<0.001), MLR (P=0.001), and SII (P=0.024) when anti-PD-1 treatment failed. Multivariate Cox regression analysis indicated that NLR (P<0.001, P=0.002), lymph node metastasis (P=0.013, P=0.001), Eastern Cooperative Oncology Group Performance Status (ECOG PS) (P=0.008, P=0.002), and treatment lines (P=0.037, P=0.048) were significant prognostic indicators of PFS and OS. Additionally, SII (P=0.016) was also significantly related to OS in ESCC patients. The risk score model showed that low risk patients prolonged PFS and OS than those with middle or high risk (P<0.001, P<0.001). Inflammatory markers can reflect short-term outcomes of anti-PD-1 treatment for ESCC patients. NLR, lymph node metastases, ECOG PS, and treatment lines are significant prognostic indicators for PFS and OS. And the risk score model constructed based on the above factors has favourable prognostic predictive value.

Expression of Polyadenylate-binding Protein Cytoplasmic 1 (PABPC1) in Combination With RAD51 as Prognostic Biomarker in Patients Who Underwent Postoperative Chemotherapy for Esophageal Squamous Cell Carcinoma.

Molecular markers in the prognosis of esophageal squamous cell carcinoma (ESCC) patients who received postoperative treatments are lacking. This research aims to evaluate the prognostic value of polyadenylate-binding protein cytoplasmic 1 (PABPC1) alone and in combination with RAD51 in ESCC patients who underwent postoperative chemotherapy (CT). A total of 103 ESCC patients who underwent postoperative CT and 103 matched ones who received surgery alone were analyzed in this study. PABPC1 and RAD51 expression was assessed in cancer samples by immunohistochemistry. PABPC1 high expression (PABPC1-HE) but not that of RAD51 was associated with poor patients' survival, regardless of the postoperative treatment or node status. Patients with PABPC1 low expression and RAD51 negative expression [RAD51- (PABPC1-LE/RAD51-)] tumor had good overall survival (OS) in both the CT treated and untreated groups. Patients with PABPC1-LE/RAD51+ and PABPC1-HE/RAD51+ tumors had longer OS in the CT treated group than in the untreated group. However, PABPC1-HE/RAD51- was associated with a poor outcome in both groups and the patients with PABPC1-HE/RAD51- tumor had hardly any benefit from CT in N+ status. PABPC1 alone and in combination with RAD51 was a prognostic biomarker for OS in ESCC patients who received postoperative CT.

Family history of cancer is a prognostic factor for better survival in operable esophageal squamous cell carcinoma: A propensity score matching analysis.

Patients with a family history of cancer, especially digestive tract cancer and esophageal cancer, a family history of cancer in the first degree, and more than one relative affected by cancer were associated with favorable survival when compared to those without a family history of cancer. A family history of cancer is a favorable independent prognostic factor in ESCC. Patients with a family history of cancer, especially digestive tract cancer and esophageal cancer, a family history of cancer in the first degree, and more than one relative affected by cancer were associated with favorable survival when compared to those without a family history of cancer. A family history of cancer (FH) is closely associated with the risk and survival of many cancers. However, the effect of FH on the prognosis of patients with esophageal squamous cell carcinoma (ESCC) remains unclear. We performed a large cohort study in the Chinese population to obtain insight into the prognostic value of FH in patients with operable ESCC. A total of 1,322 consecutive patients with thoracic ESCC who had undergone esophagectomy between January 1997 and December 2013 were included. The FH group included patients with any degree of FH, while the non-FH group included patients without any degree of FH. In total, 215 patients with FH and 215 without FH were matched using the propensity score matching analysis method to adjust for differences in baseline variables between the two groups. The impact of FH on disease-free survival (DFS) and overall survival (OS) was estimated using the Kaplan-Meier method and Cox's proportional hazards models. Before matching, 280 (21.2%) patients were included in the FH group and 1,042 (78.8%) in the non-FH group. FH was associated with early pathological T stage (p = 0.001), lymph node-negative status (p = 0.022), and early pathological stage (p = 0.006). After matching, FH was an independent prognostic factor for DFS and OS in ESCC patients. Patients with FH had 35% lower risk of disease progression (hazard ratio [HR] = 0.65, 95% CI: 0.51-0.84, p = 0.001) and 34% lower risk of death (HR = 0.66, 95% CI: 0.51-0.86, p = 0.002) than those without FH. Patients with a family history of digestive tract cancer (FH-DC), a family history of esophageal cancer (FH-EC), FH in first-degree relatives (FH-FD), and more than one relative affected by cancer were associated with favorable DFS and OS as compared to those without FH. FH is a favorable independent prognostic factor in ESCC. Patients with FH, especially those with FH-DC, FH-EC, FH-FD, and more than one relative affected by cancer, had improved survival.

Post-treatment serum triglyceride: An effective biomarker for body fat mass and overall survival in esophageal squamous cell cancer patients treated with chemoradiotherapy.

Although lipids have been assessed for their possible roles in cancer survival prediction, studies on the association between serum triglyceride (TG) levels and the prognosis of esophageal squamous cell carcinoma (ESCC) patients are limited. This study aimed to evaluate whether serum TG is associated with outcomes in patients with ESCC and investigate any interaction between serum TG and clinical parameters, especially body fat mass. We conducted a prospective case study on patients diagnosed with ESCC between March 2012 and November 2018. We measured patients' serum TG levels before and after treatment. The association between serum TG and overall survival (OS) was evaluated using hazard ratios. We sought to determine a threshold point using optimal stratification. Survival analysis was performed using Kaplan-Meier curves and a Cox proportional hazards model. Of the 257 participants diagnosed with ESCC, 200 (77.8%) were men. Median follow-up time was 22.4 months (range 3.3-92.4 months). Using univariate Cox proportional hazard analysis and subsequent multivariate analysis, post-TG levels, Karnofsky performance scores, T stages, and chemotherapy cycles were shown to be independent prognostic factors for OS (p < 0.05). The post-TG cut-off point to best classify patients with respect to time to mortality was 1.47 mmol/L. A post-TG level of ≥ 1.47 mmol/L could independently predict a better OS (hazard ratio: 0.55, 95% confidence interval: 0.37-0.79). The associations were consistent across the subtypes of clinical parameters. Furthermore, the post-body mass index, post-subcutaneous adipose tissue area, post-visceral adipose tissue area, post-total adiposity tissue area, and post-total adipose density exhibited a strong positive association with post-TG levels. Post-TG levels were found to be a significant positive prognostic biomarker for body fat mass and OS in ESCC patients.

Neoadjuvant Chemoradiotherapy vs. Neoadjuvant Chemotherapy for the Treatment of Esophageal Squamous Cell Carcinoma: A Stabilized Inverse Probability of Treatment Weighting Analysis

<h3>Purpose/Objective(s)</h3> To compare long-term survival and safety of neoadjuvant chemotherapy (nCT) and neoadjuvant chemoradiotherapy (nCRT) in patients with esophageal squamous cell carcinoma (ESCC). <h3>Materials/Methods</h3> In 2012ཞ2020, patients diagnosed with ESCC were documented in the National Cancer Center database. as follows: 1) 18-75 years old; 2) histologically proven as stage T1-2N+M0 or T3-4N0-3M0; 3) histologic subtype was ESCC; 4) R0 resection; 5) radiotherapy dose ≥ 40Gy for primary tumor and metastatic lymph nodes. Stabilized inverse probability of treatment weighting (sIPTW) were used to minimize bias due to potential confounding factors, including sex, age, tumor location, tumor length, TNM clinical stage. The outcome included overall survival (OS), disease-free survival (DFS), pathological complete responses (pCR), and perioperative mortality and morbidity. <h3>Results</h3> A total of 419 patients (male,8 7.1%; mean age, 59.7 years) were enrolled to the nCRT group (n=158) or the nCT group (n=261). median follow-up time was 26 (7ཞ113) months in the nCRT group and 24 (7ཞ101) months in the nCT group, respectively. The chemotherapy regimen of nCT group was paclitaxel combined with platinum every 21 days with a median cycle of 3 (1ཞ4). In nCRT group, paclitaxel and platinum was performed weekly with a median of 3 cycles. The median radiotherapy dose for primary tumor and metastatic lymph nodes of 45Gy (40ཞ50.4Gy), 1.8ཞ2.14Gy/fraction, was delivered by intensity-modulated radiotherapy (IMRT) or volumetric intensity modulated arc therapy (VMAT). nCRT significantly improved the pCR rate as compared to nCT group (28.5% vs 9.2%, <i>P</i><0.001). The 1, 3 and 5-year OS in the nCRT and nCT group were 96.2% vs 98.1%, 78.4% vs 70.3% and 67.9% vs 53.3%, respectively (<i>P</i> =0.140). The incidence of anastomotic leak was in 13 of 158 patients (8.2%) in the nCRT group and 4 of 261 patients (1.5%) in the nCT group (<i>P</i> The total postoperative complications rate was 10.1% in the nCRT group (16 of 158) and 7.7% in the nCT group (20 of 261). After sIPTW, the clinical and demographic characteristics of the two groups were well balanced. The nCRT group was associated with a better OS than the nCT group (96.7% vs 97.6% at 1 year; 81.0% vs 69.2% at 3 years and 72.6% vs 48.2% 5 years, <i>P</i>=0.04). <h3>Conclusion</h3> Compared to nCT, paclitaxel and platinum-based nCRT using IMRT or VMAT technique improved OS in ESCC patients. Prospective head-to-head comparison and novel strategies with advanced drug therapy and radiotherapy technique to improve the outcomes in resectable ESCC are warranted.

LncRNA RPL34-AS1 suppresses the proliferation, migration and invasion of esophageal squamous cell carcinoma via targeting miR-575/ACAA2 axis.

BackgroundLong noncoding RNAs (lncRNAs) are abnormally expressed in a broad type of cancers and play significant roles that regulate tumor development and metastasis. However, the pathological roles of lncRNAs in esophageal squamous cell carcinoma (ESCC) remain largely unknown. Here we aimed to investigate the role and regulatory mechanism of the novel lncRNA RPL34-AS1 in the development and progression of ESCC.MethodsThe expression level of RPL34-AS1 in ESCC tissues and cell lines was determined by RT-qPCR. Functional experiments in vitro and in vivo were employed to explore the effects of RPL34-AS1 on tumor growth in ESCC cells. Mechanistically, fluorescence in situ hybridization (FISH), bioinformatics analyses, luciferase reporter assay, RNA immunoprecipitation (RIP) assay and western blot assays were used to detect the regulatory relationship between RPL34-AS1, miR-575 and ACAA2.ResultsRPL34-AS1 was significantly down-regulated in ESCC tissues and cells, which was negatively correlated with overall survival in ESCC patients. Functionally, upregulation of RPL34-AS1 dramatically suppressed ESCC cell proliferation, colony formation, invasion and migration in vitro, whereas knockdown of RPL34-AS1 elicited the opposite function. Consistently, overexpression of RPL34-AS1 inhibited tumor growth in vivo. Mechanistically, RPL34-AS1 acted as a competing endogenous RNA (ceRNA) of miR-575 to relieve the repressive effect of miR-575 on its target ACAA2, then suppressed the tumorigenesis of ESCC.ConclusionsOur results reveal a role for RPL34-AS1 in ESCC tumorigenesis and may provide a strategy for using RPL34-AS1 as a potential biomarker and an effect target for patients with ESCC.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12885-022-10104-6.

Genetic variant of ADH1C for predicting survival in esophageal squamous cell cancer patients who underwent postoperative radiotherapy.

Background: Single nucleotide polymorphisms (SNPs) of essential enzymes for alcohol metabolism ADH1B, ADH1C, and ALDH2 are commonly regarded as genetic biomarkers for esophageal squamous cell carcinoma (ESCC) susceptibility. However, there have not been any reports on relations between SNPs of these genes and the prognosis of postoperative radiotherapy in ESCC. The current study aimed to understand the associations between gene variants of alcohol metabolism and adjuvant radiotherapy’s prognosis in ESCC. Methods: This study retrospectively analyzed 110 ESCC patients from our institution who received adjuvant radiotherapy after surgery. The SNPs of ADH1B rs1229984, ADH1C rs1789924, and ALDH2 rs671 were detected by Sanger sequencing using formalin-fixed paraffin-embedded tumor samples. A nomogram was drawn based on prognostic factors associated with overall survival (OS). Results: ADH1C rs1789924 (C>T) was associated with poor DFS and OS in ESCC patients undergoing adjuvant radiotherapy. Multivariate analysis showed that ADH1C rs1789924 (C>T) was one of the independent prognosis factors of DFS and OS. However, the genotypes of ADH1B SNP rs1229984 and ALDH2 rs671 were not associated with differences in the PFS and OS of these patients. Compared with the AJCC staging system, the nomogram containing the ADH1C genotype can more effectively and accurately predict the survival time of ESCC after surgery and adjuvant radiotherapy. Conclusion: ADH1C rs1789924 might be a prognostic genetic biomarker for ESCC patients undergoing surgery and postoperative radiotherapy.

MIR548P and TRAV39 Are Potential Indicators of Tumor Microenvironment and Novel Prognostic Biomarkers of Esophageal Squamous Cell Carcinoma.

Esophageal squamous cell carcinoma (ESCC) remains a common aggressive malignancy in the world. Multiple studies have shown evidence to support the hypothesis that certain functional genes that are engaged in the microenvironment of tumors played a role in the progression of ESCC. Thus, to better analyze the prognostic values of important genes in ESCC, there is an immediate need for an in-depth research study. From the TCGA database, the RNA-seq data and clinical features of 163 ESCC patients were obtained. Using the ESTIMATE technique, we were able to calculate the ImmuneScore, the StromalScore, and the ESTIMATEScore for each ESCC sample. The samples from the ESCC were split up into high score and low score groups based on the median of the various scores. In this study, ImmuneScore, StromalScore, and ESTIMATEScore were not found to be linked with overall survival of ESCC patients, according to our findings. Higher StromalScores were linked to more advanced T stages and clinical stages. The intersection analysis that was exhibited by the use of a Venn diagram indicated that there was a total of 944 upregulated genes that shared the same high score in both the ImmuneScore and the StromalScore and that there was 0 downregulated gene that shared the same low score. Survival experiments confirmed MIR548P and TRAV39 as critical prognostic biomarkers for ESCC patients. Importantly, we found that TRAV39 expression was positively associated with T cell CD4 memory activated while negatively associated with B cell memory, dendritic cells activated, and mast cells activated. In addition, we found that MIR548P expression was negatively associated with mast cells activated while positively associated with T cell CD4 memory activated. Overall, we identified MIR548P and TRAV39 as new modulators for ESCC, affecting the immune microenvironment of ESCC patients and may be a target of immunotherapy.

The circRNA circADAMTS6 promotes progression of ESCC and correlates with prognosis

Circular RNAs (circRNAs) are a type of noncoding RNA, which play a vital role in the occurrence and development of esophageal squamous cell carcinoma (ESCC). While the role of novel circADAMTS6 in ESCC remains unknown. We assessed circADAMTS6 expression in ESCC tissues and cells, and the relationship between circADAMTS6 expression and overall survival of ESCC patients. Functional experiments in vitro and xenograft in vivo assay were applied to explore the functions and mechanisms of circADAMTS6 in ESCC. Results found that up-regulation of circADAMTS6 was associated with poor overall survival and may acted as an independent risk factor for ESCC prognosis. Knockdown of circADAMTS6 significantly inhibited the proliferation, migration and invasion of ESCC cells and growth of xenograft tumors in vivo. Induced AGR2 expression was able to rescue the loss of function induced by si-circADAMTS6 in KYSE150 cell. CircADAMTS6 may acts as oncogene by activating AGR2 and the Hippo signaling pathway coactivator YAP in ESCC.

Abstract 5981: Single cell RNA sequencing uncovers high-plasticity cell state during tumor metastasis

Abstract Cancer metastasis remains the key barrier for patient’s cure. Tumor metastasis initiating from a single disseminated cell into the invasive nodule in the foreign microenvironment is poorly understand. In this study, we established the intravenous injection pulmonary metastatic model followed by single-cell RNA sequencing of disseminating cells from mice lungs at different timepoints, we identified a cluster of SLPI-high esophageal squamous cell carcinoma (ESCC) cells that mediates the lung metastasis evolution and predicts worse overall survival of ESCC patients. GSEA analysis reveals that the SLPI-high subset is functionally associated with epithelial-to-mesenchymal transition and neutrophil activation during lung metastasis. Immune infiltration prediction from TIMER also confirmed that high SLPI is tightly correlated with more abundance of neutrophils in the ESCC tumor microenvironment. SLPI is reported to drive the metastasis progression through regulating the vascular mimicry. In the next step, further in-vitro and in-vivo studies is required to explore the role of SLPI in ESCC cells in promoting lung metastasis evolution. We identified a SLPI+ ESCC cells with high pluripotency that particularly initiates the metastasis process. Citation Format: Yu Zhang, Ching Ngar Wong, Lanqi Gong, Xin-yuan Guan, Beibei Ru, Beilei Liu. Single cell RNA sequencing uncovers high-plasticity cell state during tumor metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5981.

CircFAM120B functions as a tumor suppressor in esophageal squamous cell carcinoma via the miR-661/PPM1L axis and the PKR/p38 MAPK/EMT pathway

Extensive changes of circRNA expression underscore their essential contributions to multiple hallmarks of cancers; however, their functions and mechanisms of action in esophageal squamous cell carcinoma (ESCC) remain undetermined. Here, we adopted a three-stage approach by first screening for significantly differentially expressed circRNAs in ESCC and performing an external validation study, followed by the functional analyses. The properties of circRNAs were evaluated using Sanger sequencing, RNase R digestion, actinomycin D treatment, subcellular localization analysis, and fluorescence in situ hybridization. Target transcripts were predicted using online tools and verified by dual-luciferase, RNA immunoprecipitation, qRT-PCR, and western blot. Biotin-labeled RNA-protein pull-down, mass spectrometry, and RNA immunoprecipitation were employed to identify proteins interacting with circRNAs. Gain- and loss-of-function experiments were performed to uncover the roles of circRNAs, their target genes, and binding proteins in the proliferation, metastasis, and invasion. We observed that circFAM120B (hsa_circ_0001666) was frequently downregulated in cancer tissues and patient plasma, and its expression level was related to overall survival in ESCC patients. Overexpression of circFAM120B inhibited the proliferation, metastasis, and invasion of ESCC while silencing it enhanced malignant phenotypes. Mechanistically, circFAM120B was predominantly located in the cytoplasm, guarantying its sponging for miR-661 to restore the expression of PPM1L, a tumor suppressor. We observed that circFAM120B could reduce the stability of RNA-dependent protein kinase (PKR) by promoting its ubiquitination-dependent degradation and subsequently regulating the p38 MAPK signaling pathway, resulting in the repression of EMTs in ESCC cells. Our findings suggest that circFAM120B is a promising biomarker of ESCC, which acts as a tumor suppressor via the circFAM120B/miR-661/PPM1L axis and PKR/p38 MAPK/EMT pathway, supporting its significance as a candidate therapeutic target.

Computed Tomography-Based Radiomics Nomogram for Predicting the Postoperative Prognosis of Esophageal Squamous Cell Carcinoma: A Multicenter Study

The aim of this study was to evaluate and identify the predictive value of combining CT radiomics features and clinical features to determine recurrence-free survival (RFS) and overall survival (OS) after surgery in patients with esophageal squamous cell carcinoma (ESCC). A total of 372 patients with surgically and pathologically confirmed ESCC from 2 institutions were retrospectively included. All patients from institution 1 were randomized at a 7:3 ratio into a training cohort (n=206) and an internal validation cohort (n=88), and patients from institution 2 were used as an external validation cohort (n=78). The association between the radiomics features and RFS and OS was assessed in the training cohort and verified in the validation cohort. Furthermore, the performance of the radiomics nomogram was evaluated by combining the radiomics score (rad-score) and clinical risk factors. The radiomics nomogram that combined radiomics features and clinical risk factors was better than the clinical nomogram and radiomics model alone at predicting RFS and OS in ESCC patients. All calibration curves showed significant consistency between predicted survival and actual survival. Radiomics features could be used to stratify patients with ESCC following radical resection into high- and low-risk groups. Furthermore, the radiomics nomograms provided better predictive accuracy than other predictive models and might serve as a therapeutic decision-making reference for clinicians and be used to monitor the risks of recurrence and death.

Increased Drp1 promotes autophagy and ESCC progression by mtDNA stress mediated cGAS-STING pathway

BackgroundMitochondrial dynamics homeostasis is important for cell metabolism, growth, proliferation, and immune responses. The critical GTPase for mitochondrial fission, Drp1 is frequently upregulated in many cancers and is closely implicated in tumorigenesis. However, the mechanism underling Drp1 to influence tumor progression is largely unknown, especially in esophageal squamous cell carcinoma (ESCC).MethodsImmunohistochemistry was used to examine Drp1 and LC3B expression in tissues of ESCC patients. Autophagic vesicles were investigated by transmission electron microscopy. Fluorescent LC3B puncta and mitochondrial nucleoid were observed by fluorescent and confocal microscopy. Mitochondrial function was evaluated by mitochondrial membrane potential, ROS and ATP levels. Xenograft tumor model was performed in BALB/c nude mice to analyze the role of Drp1 on ESCC progression.ResultsWe found that Drp1 high expression is correlated with poor overall survival of ESCC patients. Drp1 overexpression promotes cell proliferation and xenograft ESCC tumor growth by triggering autophagy. Furthermore, we demonstrated that Drp1 overexpression disturbs mitochondrial function and subsequent induces mitochondrial DNA (mtDNA) released into the cytosol thereby inducing cytosolic mtDNA stress. Mechanistically, cytosolic mtDNA activates the cGAS-STING pathway and facilitates autophagy, which promotes ESCC cancer growth. Moreover, mtDNA digestion with DNase I and autophagy inhibition with chloroquine attenuates the cGAS-STING pathway activation and ESCC cancer growth.ConclusionsOur finding reveals that Drp1 overexpression induces mitochondrial dysfunction and cytosolic mtDNA stress, which subsequently activates the cGAS-STING pathway, triggers autophagy and promotes ESCC progression.

Correlation between TXNRD1/HO-1 expression and response to neoadjuvant chemoradiation therapy in patients with esophageal squamous cell carcinoma

BackgroundThioredoxin reductase 1 (TXNRD1) and heme oxygenase-1 (HO-1) are both involved in the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway and play key roles in antioxidant responses. In patients with esophageal squamous cell carcinoma (ESCC), the correlation between the expression of these two proteins and the therapeutic response to neoadjuvant chemoradiation therapy (NACRT), as well as the difference in their expression after chemoradiotherapy, remains unknown.MethodsProteins involved in the Nrf2 pathway were immunolocalized in carcinoma cells in ESCC patients on NACRT with 5-fluorouracil and cisplatin, followed by esophagectomy. The 8-hydroxydeoxyguanosine (8-OHdG) levels were used to quantify reactive oxygen species. The changes in immunoreactivity before and after NACRT (Δ) were assessed.ResultsTumor reduction following NACRT was significantly attenuated in pre-therapeutic biopsy specimens associated with high HO-1 status. TXNRD1Δ, HO-1Δ, and 8-OHdGΔ were significantly different in the ineffective and effective groups. The overall survival was significantly lower in high Nrf2 and TXNRD1 groups. In addition, high TXNRD1 expression was an independent prognostic factor in the multivariate analysis of overall survival.ConclusionsThe study findings indicate that HO-1 status in pre-therapeutic biopsy specimens could predict response to NACRT, and TXNRD1 status could predict overall survival of ESCC patients.