Esophageal Intraepithelial Neoplasia Research Articles

Epigenetic silencing of JAM3 promotes esophageal cancer development by activating Wnt signaling

BackgroundThe role of JAM3 in different tumors is controversial. The epigenetic regulation and the mechanism of JAM3 remain to be elucidated in human esophageal cancer (EC).MethodsEleven EC cell lines, 49 cases of esophageal intraepithelial neoplasia (EIN) and 760 cases of primary EC samples were employed. Methylation-specific polymerase chain reaction, immunohistochemistry, MTT, western blot and xenograft mouse models were applied in this study.ResultsThe inverse association between RNA expression and promoter region methylation of JAM3 was found by analyzing 185 cases of EC samples extracted from the TCGA database (p < 0.05). JAM3 was highly expressed in KYSE450, KYSE520, TE1 and YES2 cells, low level expressed in KYSE70 cells and unexpressed in KYSE30, KYSE150, KYSE410, KYSE510, TE13 and BIC1 cells. JAM3 was unmethylated in KYSE450, KYSE520, TE1 and YES2 cells, partial methylated in KYSE70 cells and completely methylated in KYSE30, KYSE150, KYSE410, KYSE510, TE13 and BIC1 cells. The expression of JAM3 is correlated with methylation status. The levels of JAM3 were unchanged in KYSE450, KYSE520, TE1 and YES2 cells, increased in KYSE70 cells and restored expression in KYSE30, KYSE150, KYSE410, KYSE510, TE13 and BIC1 cells after 5-aza-2′-deoxycytidine treatment, suggesting that the expression of JAM3 is regulated by promoter region methylation. JAM3 was methylated in 26.5% (13/49) of EIN and 51.1% (388/760) of primary EC, and methylation of JAM3 was associated significantly with tumor differentiation and family history (all p < 0.05). Methylation of JAM3 is an independent prognostic factor of poor 5-year overall survival (p < 0.05). JAM3 suppresses cell proliferation, colony formation, migration and invasion and induces G1/S arrest and apoptosis in EC. Further study demonstrated that JAM3 suppressed EC cells and xenograft tumor growth by inhibiting Wnt/β-catenin signaling.ConclusionJAM3 is frequently methylated in human EC, and the expression of JAM3 is regulated by promoter region methylation. JAM3 methylation is an early detection and prognostic marker of EC. JAM3 suppresses EC growth both in vitro and in vivo by inhibiting Wnt signaling.

Proteomic profiling of plasma exosomes reveals CD82 involvement in the development of esophageal squamous cell carcinoma

The Xinjiang Uygur autonomous region has a high incidence of esophageal cancer. For the early diagnosis of patients with esophageal squamous cell carcinoma (ESCC), exosomes were isolated and quantified by liquid chromatography tandem mass spectrometry ((LC–MS/MS) with data independent acquisition (DIA) from the peripheral blood of patients with benign esophageal disease (BED), esophageal intraepithelial neoplasia (EIN) and ESCC. A total of 1117 proteins were identified in the above 9 samples. The proteomic results showed that the quantity of CD82 in exosomes of EIN was significantly higher than that in patients with BED and ESCC. Meanwhile, our ELISA test verified our proteomic results. In addition, the immunohistochemical results showed high CD82 expression in adjacent normal tissues and low expression in ESCC tissues. CD82 expression in ESCC tissues was negatively correlated with tumor stage and the expression of PKM2, and the high expression of CD82 combined with low expression of PKM2 in ESCC tissues suggested a good prognosis. To further clarify the tumor suppressive mechanism of CD82, the TIMER and TISDB databases were analyzed, and CD82 expression in tumor tissues was found to be related to the infiltration of immune cells. CD82 in exosomes is involved in the development of ESCC. SignificanceXinjiang is a high incidence area of ESCC. When diagnosed in the middle and late stages of the disease, the prognosis of patients is poor. Exosomes provide the possibility of relatively noninvasive and early detection of esophageal carcinogenesis. To the best of our knowledge, this was the first study using the DIA technique to analyze the exosomal proteins of patients with different stages of ESCC. The proteins identified in the exosomes in these three groups could provide insights for understanding how exosomes promote the occurrence of ESCC, the antitumour mechanism of humans and the early diagnosis of ESCC.

Safety and effectiveness of mucosal traction using a snare combined with endoclips to assist the resection of esophageal intraepithelial neoplasia: a propensity score matching analysis

Currently, the reports on esophageal endoscopic submucosal dissection (ESD) assisted by traction with a snare are rare. Because a snare is a commonly used endoscopic accessory and is easily available, its application in mucosal traction is worth exploring. The present study aims to evaluate the safety and effectiveness of snare-endoclip traction-assisted ESD for esophageal intraepithelial neoplasia. Cases of esophageal intraepithelial neoplasia resected using ESD in the Digestive Endoscopy Center of Guangzhou Nanfang Hospital, China from June 2013 to March 2019 were retrospectively analyzed. The procedure of snare-endoclip traction-assisted ESD was compared with nontraction-assisted ESD by using a propensity score matching analysis. Operation time, en bloc and R0 resection, intra- and postoperative complications, and surgery-related costs were mainly evaluated. Overall, 99 cases of esophageal intraepithelial neoplasia under tissue biopsy were included in the present study. Further, 22 exact matched pairs were obtained. There were no differences in en bloc and R0 resection rates, intra- and postoperative complications, and costs of disposable surgical accessories between the traction group and the nontraction group. However, median operation time showed a significant difference: traction group, 50.0min (range, 20-100min); nontraction group, 70.0min (range, 35-133min), P=0.012. In conclusion, snare-endoclip traction-assisted ESD for esophageal intraepithelial neoplasia was safe and shortened operation time in the study, thereby improving the efficiency of ESD. Despite the additional use of a snare and endoclips for traction, the total costs of endoscopic accessories seemed not to be increased.

Expression of ADAM29 and FAM135B in the pathological evolution from normal esophageal epithelium to esophageal cancer: Their differences and clinical significance.

A Disintegrin And Metalloprotease Domain 29 (ADAM29) and Family with sequence similarity 135 member B (FAM135B) genes have been reported to be associated with a carcinogenic risk of esophageal squamous cell carcinoma (ESCC). However, to the best of our knowledge, the expression of ADAM29 and FAM135B in the pathological evolution from normal esophageal epithelial cells to ESCC has not yet been investigated. The present study aimed to investigate the expression of ADAM29 and FAM135B in normal esophageal mucosal epithelium, low-grade and high-grade esophageal intraepithelial neoplasia, and ESCC. Furthermore, the present study aimed to investigate the role of ADAM29 and FAM135B in the development of esophageal lesions. Immunohistochemistry was performed in order to detect the expression levels of ADAM29 and FAM135B proteins in normal esophageal mucosa samples (40 cases), low-grade intraepithelial neoplasia samples (20 cases), high-grade intraepithelial neoplasia samples (20 cases) and ESCC samples (40 cases). The results of the present study demonstrated that the positive rates of ADAM29 and FAM135B proteins increased gradually from normal esophageal mucosal epithelium and esophageal intraepithelial neoplasia, to ESCC (P<0.05). Furthermore, the expression levels of ADAM29 and FAM135B proteins in ESCC were not associated with age and the tumor size (P>0.05); however, the protein levels were associated with the pathological stage, clinical stage and lymph node metastasis of ESCC (P<0.05). In addition, there was a significant association between the expression levels of ADAM29 protein and FAM135B protein (χ2=60.071; P<0.001). The results of the present study demonstrated that the expression levels of ADAM29 and FAM135B were associated with the tumor behavior characteristics and the progression of esophageal cancer, the expression of which could be used for the diagnosis of early esophageal cancer, and provide the basis for guiding individualized treatment.

Significant association between 53 BP1 expression and grade of intraepithelial neoplasia of esophagus: Alteration during esophageal carcinogenesis

BackgroundAbnormal DNA damage response (DDR) leads to genomic instability and carcinogenesis. P53-binding protein 1 (53 BP1), a DDR molecule, is known to accumulate at the sites of DNA double-strand breaks. The aim of this study was to analyze the expression pattern of 53 BP1-nuclear foci (NF) in esophageal neoplasms in order to visualize the state of DDR in esophageal carcinogenesis and to clarify its significance in the molecular pathology of the disease. MethodsA total of 61 lesions from 22 surgically resected samples of esophageal cancer, including histologically normal squamous epithelium, low-grade intraepithelial neoplasia (LG-IN), high-grade intraepithelial neoplasia (HG-IN), carcinoma in situ (CIS), and invasive squamous cell carcinoma (SCC), were included in the study. 53 BP1 and Ki-67 expression were analyzed by double-labeled immunofluorescence. ResultsThe number of discrete 53 BP1-NF increased as the tumor progressed from normal epithelium through LG-IN, HG-IN, CIS, and SCC. 53 BP1-NF larger than 1 μm in diameter (large foci), indicating intensive DDR, also showed a stepwise increase during the progression of carcinogenesis. Of note, large foci of 53 BP1 were found in significantly higher numbers in HG-IN than in LG-IN. Furthermore, localization of 53 BP1-NF in Ki-67-positive cells, indicating the abnormal timing of DDR, also increased with malignancy progression. Conclusions53 BP1-NF accumulation increases during cancer progression from LG-IN to HG-IN to CIS to SCC. Detection of 53 BP1-NF by immunofluorescence, especially large foci, is a feasible method of estimating DNA instability and the malignant potential of esophageal intraepithelial neoplasia.

Expressions and significance of the second mitochondria-derived activator of caspase protein and Survivin in tissues of high-grade intraepithelial neoplasia and esophageal squamous cell carcinoma

Objective To detect the expressions of the second mitochondria-derived activator of caspase protein(Smac) and Survivin in tissues of high-grade intraepithelial neoplasia(HGIN)and esophageal squamous cell carcinoma (ESCC), and explore the influence of Smac and Survivin on occurrence and development of ESCC. Methods The retrospective case-control study was conducted. The clinicopathological data of 45 healthy controls with suspected esophagitis (control group), 41 patients with HGIN (HGIN group) and 57 patients with ESCC (ESCC group) who were admitted to the Qinghai University Affiliated Hospital between January 2012 and December 2014 were collected. Observation indicators included: (1) expressions of Smac and Survivin were detected by immunohistochemistry. (2) Expressions of mRNA of Smac and Survivin in tumor tissues and adjacent tissues were detected by real-time polymerase chain reaction (RT-PCR). (3) Correlation between relative expressions of Smac and Survivin in tumor tissues and adjacent tissues. (4) Relationship between expressions of Smac and Survivin in tumor tissues and clinicopathological factors. Measurement data with normal distribution were represented as ±s. The comparisons among groups was evaluated with the one-way ANOVA. The comparison between tumor tissues and adjacent tissues were analyzed by the paired t test, and comparison of rates was done using the chi-square test and Fisher exact probability. The correlation between expressions of Smac and Survivin was done using the Pearson correlation analysis. Results (1) Expressions of Smac and Survivin were detected by immunohistochemistry. The positive expression rate of Smac of esophageal tissues in the control group, HGIN group and ESCC group were respectively 82.2%(37/45), 43.9%(18/41) and 40.4%(23/57), with a statistically significant difference among the 3 groups (χ2=20.408, P 0.05). The positive expression rate of Smac of adjacent tissues in the ESCC group was 84.2%(48/57), with a statistically significant difference between tumor tissues and adjacent tissues (χ2=20.530, P 0.05). The positive expression rate of Survivin of adjacent tissues in the ESCC group was 15.8%(9/57), with a statistically significant difference between tumor tissues and adjacent tissues (χ2=32.386, P 0.05) and in adjacent tissues (r=-0.197, P>0.05). There was a correlation in the relative expression of Smac between tumor tissues and adjacent tissues (r=0.527, P 0.05). (4) The relationship between the expressions of Smac and Survivin in tumor tissues and clinicopathologic factors: the positive expression of Smac in tumor tissues was 51.6% in men, 9.1% in women, 64.7% in tumor diameter<3 cm, 28.0% in tumor diameter≥3 cm, 60.0% in stage Ⅰ-Ⅱ of TNM stage and 22.7% in stage Ⅲ-Ⅳ of TNM stage, respectively, showing statistically significant differences in the positive expression of Smac in tumor tissues of patients with different gender, tumor diameter and TNM stage (χ2=6.093, 5.567, 6.041, P<0.05). The positive expression of Survivin in tumor tissues was 54.5% in high- and moderate-differentiated tumors, 100.0% in low-differentiated tumors, 75.0% in stageⅠ-Ⅱ and 100.0% in stage Ⅲ-Ⅳ, respectively, showing statistically significant differences in the positive expression of Survivin in tumor tissues of patients with different tumor differentiation and TNM stage (P<0.05). Conclusions The expression of Smac in the progression of esophageal epithelial cells towards to high-grade intraepithelial neoplastia and proliferatoin is inhibited, while expression of Survivin is activated. Smac and Survivin participate in the occurrence of ESCC and maybe play an role in determining the biological features of carcinoma tissues. Key words: Esophageal Neoplasms; Second mitochondria-derived activator of caspase protein; Survivin protein; High-grade esophageal intraepithelial neoplasia

FOXO1 Downregulation Correlates with Progression of Esophageal Intraepithelial Neoplasia

Background: Forkhead transcription protein 1 (FOXO1) is an important transcriptional regulator of cell proliferation and is considered essential for tumor growth and progression. However, the function of FOXO1 in esophageal cancer remains unclear. Esophageal squamous intraepithelial neoplasia (ESIN) has been widely recognized as a precursor lesion for esophageal squamous cell carcinoma (ESCC). Early detection offers the best prognosis for esophageal squamous cell carcinoma. The differentiation of squamous dysplasia from reactive change and the classification of ESIN into high-grade (HGIEN) or low-grade (LGIEN) are sometimes subjective and challenging. In this study, we sought to evaluate the FOXO1 expression in endoscopically biopsied esophageal cases and compare its expression with those of p AKT1, Ki67 and p53 to detect where is the lesion stand in the scale from normal mucosa to HGIEN in order not to miss prone cancer cases. Methodology and results: Immunohistochemical staining was performed on 99 formalin-fixed, paraffin embedded blocks of endoscopic esophageal samples. Out of them, 44 samples were ESIN and 33 samples were ESCC, further, for comparison, 22 samples of reactive hyperplastic, non-dysplastic esophageal mucosa. The current study showed a significant stepwise decrease in the FOXO1 expression as well as increased expression of p AKT1, ki67 and P53 from the progression of LGIEN to HGIEN to ESCC. HGIEN and ESCC strongly associated with low FOXO1 expression. On the other hand, higher expression of p AKT1, ki67 and p53 was more associated with HGIEN and ESCC (all are P: 0.001). Conclusions: This study demonstrated stepwise under-expression of FOXO1 in the progression of esophageal squamous carcinogenesis so that immunohistochemical assessment of this marker might provide a useful adjunct tool for differential diagnosis between LGIEN and HGIEN as well as could be used in targeting therapy.

Expression and role of oncogenic miRNA-224 in esophageal squamous cell carcinoma.

BackgroundAberrant expression of miR-224 is associated with tumor development and progression. This study investigated the role of miR-224 in esophageal squamous cell carcinoma (ESCC) ex vivo and in vitro.MethodsA total of 103 esophageal intraepithelial neoplasia, ESCC tissue specimens, and their matched distant normal tissues were collected to test miR-224 expression using qRT-PCR analysis. Western blot was used to quantify the level of PH domain leucine-rich repeat protein phosphatase 1 (PHLPP1) and PHLPP2 in ESCC tissues. Cell viability, apoptosis, invasion, and colony formation assays were used to assess the altered phenotypes of esophageal cancer cell lines after miR-224 expression or inhibition. A luciferase reporter assay was used to confirm miR-224 binding to PHLPP1 and PHLPP2 mRNA.ResultsmiR-224 was significantly overexpressed in esophageal intraepithelial neoplasia and ESCC tissues, while the expression of PHLPP1 and PHLPP2 proteins, the target genes of miR-224, was downregulated in ESCC tissues. miR-224 expression was associated with advanced clinical TNM stage, pathologic grade, and the level of PHLPP1 and PHLPP2 proteins in ESCC tissues. Ectopic overexpression of miR-224 promoted proliferation, migration, and invasion, but suppressed apoptosis of ESCC cells. miR-224 was able to bind to the 3′ untranslated region (3′-UTR) of PHLPP1 and PHLPP2 mRNA to suppress their expression.ConclusionsThe current study demonstrated that miR-224 acts as an oncogenic miRNA in ESCC, possibly by targeting PHLPP1 and PHLPP2.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-015-1581-6) contains supplementary material, which is available to authorized users.

Clinical efficacy of multi-band mucosectomy for high-grade esophageal intraepithelial neoplasia

Objective To investigate the efficacy and safety of multi-band mucosectomy (MBM) for the treatment of high-grade intraepithelial neoplasia.Methods The clinical data of 24 patients with high-grade esophageal intraepithelial neoplasia who were admitted to the Henan Honliv Hospital from April 2011 to October 2012 were retrospectively analyzed.All the 24 patients received MBM,and the operation time,resection of the lesion and complications were observed.All the patients were followed up with gastroscopy at postoperative 1,3,6,12 months.The follow-up was ended in April 2013.Results A total of 26 lesions in 24 patients were resected successfully by MBM (2 patients received MBM twice).The mean operation time was 42 minutes.The mean length of the lesions was 3.1 cm (range,0.8-6.0 cm),and it occupied 3/4 of the circumference of the esophagus to the maximum.The mean number of the band used was 4 (range,1-6).During the operation,bleeding was occurred in 4 patients,and they were cured by hot biopsy forceps.No perforation of the esophagus was detected.The results of post-MBM pathological examination showed that 22 patients were with severe atypical hyperplasia,and 2 were with carcinoma in situ.During the follow-up of 6-24 months,22 patients were cured,and esophageal stricture occurred in 1 patient at post-MBM 1 month,and the symptoms were successfully relieved by endoscopic balloon dilatation.Neoplasia recurrence was observed in 1 patient (2 lesions were resected twice) at post-MBM 3 months,and he received surgical treatment.Histopathological diagnosis showed that he had esophageal squamous cell carcinoma.No stricture or neoplasia was detected by gastroscopy at postoperative month 12.Conclusions MBM is a relatively safe and effective endoscopic technique for the treatment of high-grade esophageal intraepithelial neoplasia.The resection range should not be blindly extended.For patients whose lesions are beyond 3/4 of the circumference of the esophagus in width or with multiple lesions which can not be resected by MBM at one time,MBM should be applied cautiously to avoid esophageal stricture and recurrence. Key words: Esophageal neoplasms; High-grade intraepithelial neoplasia; Multi-band mucosectomy; Treatment

Expression and significance of EGFR, Her-2 and TOPO II in esophageal canceration course

Objective To investigate the expression of EGFR, Her-2 and TOPO Ⅱ in esophageal canceration course, analyze the correlation between the expression and clinical pathological parameters and the correlation of the three genes. Methods EGFR, Her-2 and TOPO Ⅱ were detected by Tissue microarray technology and Envision immunohistochemistry method in 107 cases of esophageal carcinoma, including normal esophageal epithelium, esophageal intraepithelial neoplasia and esophageal squamous cell carcinoma. Results The positive expression rates of EGFR and TOPO Ⅱ display an improving trend from normal esophageal epithelium, intraepithelial neoplasia to carcinoma (P =0.031) in the above four groups. The positive expression rates of EGFR were 8.41 %, 7.94 %, 27.27 %, 50.47 %, and TOPO Ⅱ were 3.74 %, 4.76 %, 20.45 %, 43.93 %. Furthermore, the expression showed gradually incresing with histological grades advance (P =0.009). There was no correlation between EGFR or TOPO Ⅱ and gender, age, lymph node or distant metastasis (P >0.05). There was a positive correlation between EGFR and TOPO Ⅱ (r 1=0.410, P 0.05), no correlation was obtained between Her-2 and gender, age, the depth of invasion or lymph node metastasis (P >0.05), no relationship between Her-2 and EGFR or TOPO Ⅱ either. Conclusion EGFR and TOPO Ⅱ are closely related to the occurrence and development of esophageal squamous cell carcinoma, their expressions all make a qualitative change in the esophageal high-grade intraepithelial neoplasia. The role of Her-2 in the development of esophageal squamous cell carcinoma is still not definite in Shanxi province. Key words: Esophageal neoplasms; Receptor, epidermal growth factor; Human epidermal growth factor receptor 2; Topoisomerase Ⅱ

Expression and significance of CDK4,p18,p19 in canceration of esophageal epithelium

Objective To investigate the role of cell cycle regulatory protein CDK4,p18,p19 in the genesis and development of esophageal squamous cell carcinoma (SCC).Methods Tissue microarray and immunohistochemical method (Envision) were used to detect the protein expression of CDK4,p18,p19 in 120 cases of esophageal tissues.The results were statistically analyzed.Results The positive rate of CDK4 protein expression in normal esophageal epithelium was low [28.3 % (34/120)],it increased in esophageal intraepithelial neoplasia [32.5 % (39/120)],and it was high in esophageal SCC [84.2 % (101/120)],which increased with the degree of SCC differentiation decreasing gradually.There was significant differences between the SCC and normal esophageal epithelium or esophageal intraepithelial neoplasia (x2= 76.004,P ＜0.05; x 2= 65.897,P ＜ 0.05).The expression of CDK4 in group with lymphatic metastasis [93.88 % (46/49)]was higher than without it [71.43 % (55/71)] (x2= 5.860,P ＜ 0.05).The positive rates of p18,p19 protein expression in normal esophageal epithelium were high [34.2 % (41/120),29.2 % (35/120)],it decreased in esophageal intraepithelial neoplasia [19.2 % (23/120),15.0 % (1 8/120)] (x 2= 134.481,P ＜ 0.05; x 2 = 141.376,P ＜ 0.05),but it were high in esophageal SCC [63.3 % (76/120) and 61.7 % (74/120)] which decreased with the degree of SCC differentiation gradually increased.There were significant differences between the normal esophageal epithelium and esophageal intraepithelial neoplasia,esophegeal intraepithelial neoplasia and SCC,normal esophageal epithelium and SCC (p 18:x 2 = 6.903,48.296,20.429,P ＜ 0.05; p1 9:x2 = 6.998,55.276,25.565,P＜ 0.05).CDK4 protein expression was correlated with both p18 and p19 (r =0.696,0.630,P ＜0.05),and there was significant positive correlation between the protein expression of p18 and p19 (r =0.833,P ＜0.05).Conclusion Cell cycle regulatory gene CDK4,p18,p19 get involved in the genesis and development of esophageal squamous cell carcinoma.Their protein expressions are closely related to canceration of esophageal epithelium. Key words: Esophageal neoplasms; Neoplasms,squamous cell; Cell cycle proteins; Immunohistrochemistry; p18; p19

Clinical symptoms of esophageal intraepithelial neoplasia: an analysis of 101 cases

Clinical symptoms of esophageal intraepithelial neoplasia: an analysis of 101 cases

Squamous esophageal histology and subsequent risk of squamous cell carcinoma of the esophagus.A prospective follow-up study from Linxian, China

Linxian, China, has some of the highest rates of esophageal cancer in the world. Previous authors have proposed that esophagitis, atrophy, and dysplasia may be precursor lesions of esophageal cancer in such high risk populations. To examine the relationship between squamous esophageal histology and subsequent esophageal cancer in Linxian, the authors prospectively followed 682 participants of a 1987 endoscopic survey for 3.5 years and compared their initial biopsy diagnoses with the occurrence of squamous cell carcinoma during this follow-up period. Squamous cell carcinoma of the esophagus was identified in 52 (7.6%) of the participants during the follow-up period. After adjusting for potential confounding factors, relative risks (95% confidence intervals) for squamous cell carcinoma incidence by initial histologic diagnoses were as follows: normal, 1.0 (reference); basal cell hyperplasia, 2.1 (0.4-9.8); mild dysplasia, 2.2 (0.7-7.5); moderate dysplasia, 15.8 (5.9-42.2); severe dysplasia, 72.6 (29.8-176.9); dysplasia not otherwise specified, 22.9 (6.7-78.0); and carcinoma in situ, 62.5 (24.1-161.9). In this study, moderate dysplasia, severe dysplasia, and carcinoma in situ were the only histologic lesions associated with a significantly increased risk of developing squamous cell carcinoma of the esophagus within 3.5 years after endoscopy. Increasing grades of dysplasia were associated with increasing risk, but severe dysplasia were associated with increasing risk, but severe dysplasia and carcinoma in situ had similar degrees of risk, findings that suggest a continuous spectrum of esophageal intraepithelial neoplasia, without morphologically distinguishable dysplasia and in situ carcinoma. A longer follow-up will be necessary to fully evaluate the less severe diagnostic categories, which may take more than 3.5 years to affect the occurrence of squamous cell carcinoma in this high risk population.