Association between functional polymorphisms in the flanking region of miR-143/145 and risk of papillary thyroid carcinoma: A case-control study

MiR-143 and miR-145 were down-regulated in papillary thyroid carcinoma (PTC) involving in cell proliferation, apoptosis, migration, invasion, and epithelial to mesenchymal transition. In this study, we aimed to investigate the association between 2 functional polymorphisms (ie, rs4705342 and rs353292) in the flanking region of miR-143/145 and risk of PTC.A case-control study including 316 PTC patients and 347 controls was performed. The rs4705342 and rs353292 were genotyped by using the TaqMan allelic discrimination. The results were confirmed by DNA sequencing.For the rs4705342, a reduced risk of PTC was observed in heterozygous comparison, dominant genetic model and allele comparison (CC vs TT: adjusted OR = 0.37, 95% CI = 0.19–0.74, P = .003; CT/CC vs TT: adjusted OR = 0.64, 95% CI = 0.47–0.87, P = .005; C vs T: adjusted OR = 0.66, 95% CI = 0.52–0.85, P = .001, respectively). No significant difference was found in the genotypic distributions of the rs353292 between cases and controls.These findings indicate that the rs4705342 in the flanking region of miR-143/145 may be a protective factor against the occurrence of PTC. Further study is therefore required to investigate the correlation between the genotype and V-raf murine sarcoma viral oncogene homolog B1 V600E, rat sarcoma viral oncogene homolog mutations, rearranged in transformation/PTC1 and rearranged in transformation/PTC3.     

Micro RNA-1290 promotes esophageal squamous cell carcinoma cell proliferation and metastasis

AIM:To investigate the biological role of mi R-1290 in esophageal squamous cell carcinoma(ESCC) progression and invasion and the underlying mechanism.METHODS:Quantitative real-time polymerase chain reaction(q RT-PCR) was performed to evaluate mi R-1290 expression in ESCC tissue samples.The roles of mi R-1290 in cell proliferation,migration and invasion were identified using mi R-1290 mimic-transfected cells.In addition,the regulatory effect of mi R-1290 on suppressor of cancer cell invasion(SCAI) was evaluated using q RT-PCR,Western blot analysis and a dual luciferase reporter assay.RESULTS:mi R-1290 was significantly upregulated in ESCC tissue samples compared with normal adjacent tissues(9.213 ± 1.150 vs 1.000 ± 0.0),(P 0.01).Upregulation of mi R-1290 was associated with tumor differentiation(P = 0.021),N classification(P = 0.006) and tumor-node-metastasis stage(P = 0.021) in ESCC patients.Moreover,ectopic mi R-1290 expression potently promoted ESCC cell growth(P 0.01),migration(P 0.01) and invasion(P 0.01) in vitro.mi R-1290 overexpression in ESCC cell lines decreased SCAI expression at the translational level and reduced SCAI-driven luciferase-reporter activity(P 0.01).CONCLUSION:Our findings suggested that mi R-1290 may play an oncogenic role in cellular processes of ESCC.     

食管鳞状细胞癌类器官的应用

类器官(organoid)作为一种新型的研究模型能够稳定保持肿瘤多细胞团的异质性特征,高度还原原位肿瘤组织的生理结构和功能。既能适用于高通量的临床药物筛选,提供个性化治疗的策略;又能构建病理模型,作为研究肿瘤发生、多个阶段发展和转移机制的有力工具。目前,食管、胃、肠、肝、胰、前列腺和乳腺等结构的类器官和相应的肿瘤类器官已有报道,开拓了体外培养的新平台。肿瘤类器官模型具有易操作、成本相对低廉且可以与其他先进技术相结合应用等明显优势,有望在相关领域广泛应用。本文对食管鳞状细胞癌类器官的培养技术及应用现状进行了总结。     

Blood-based biomarkers for early detection of esophageal squamous cell carcinoma

Esophageal squamous cell carcinoma(ESCC) is a common malignant tumor of the digestive system worldwide, especially in China. Due to the lack of effective early detection methods, ESCC patients often present at an advanced stage at the time of diagnosis, which seriously affects the prognosis of patients. At present,early detection of ESCC mainly depends on invasive and expensive endoscopy and histopathological biopsy. Therefore, there is an unmet need for a noninvasive method to detect ESCC in the early stages. With the emergence of a large class of non-invasive diagnostic tools, serum tumor markers have attracted much attention because of their potential for detection of early tumors. Therefore, the identification of serum tumor markers for early detection of ESCC is undoubtedly one of the most effective ways to achieve early diagnosis and treatment of ESCC.This article reviews the recent advances in the discovery of blood-based ESCC biomarkers, and discusses the origins, clinical applications, and technical challenges of clinical validation of various types of biomarkers.     

肿瘤标志物在食管鳞状细胞癌中的研究与应用

食管鳞状细胞癌作为食管癌的主要类型, 是人类最常见的消化系恶性肿瘤之一. 作为诊断手段之一的肿瘤标志物检测, 具有简便、经济、快速、无创的特点, 更重要的是一些标志物在组织器官发生形态学变化之前就有表达, 因此, 肿瘤标志物对食管癌的研究就更有意义.本文综述近几年来一些发现和检测到的肿瘤标志物在食管鳞状细胞癌中的差异表达, 分别从基因、蛋白、自身免疫抗体、抗原及预测因子角度总结介绍.     

基因芯片技术在食管鳞癌转移相关基因表达谱筛选中的应用

目的 研究人食管鳞癌转移相关基因表达谱,探讨食管鳞癌的转移机制。方法 选取392个与肿瘤转移相关的基因克隆,制备成肿瘤转移基因芯片。提取食管鳞癌组织以及正常食管组织RNA,反转录后标记为cDNA探针,与cDNA芯片杂交,经扫描及Quantarray 3．0软件分析后比较两种组织中的差异表达基因。结果 共筛查出差异表达基因58条,其中表达上调基因36条、下调基因22条,包括癌基因、抑癌基因、黏附分子、基质金属蛋白酶、信号转导因子、细胞代谢和免疫相关基因等。结论 基因芯片筛查食管鳞癌转移相关基因表达谱可为明确食管鳞癌转移机制提供重要参考。     

Elevated serum levels of human relaxin-2 in patients with esophageal squamous cell carcinoma

AIM: To assess the prognostic value of serum human relaxin 2 (H2 RLN) level in patients with esophageal squamous cell carcinoma (ESCC). METHODS: From October 1998 to September 2009, 146 patients with histopathologically confirmed ESCC were enrolled in this study. One hundred patients underwent en bloc esophagectomy, and 46 patients with unresectable tumors underwent palliative surgery. Five of the 146 patients died of surgical complications. Serum levels of H2 RLN were measured by enzyme linked immunosorbent assay. The relationship between serum H2 RLN level and each of the clinicopathological parameters was analyzed using the χ2 test. Patients were classified into two groups according to their H2 RLN level (< 0.462 ng/mL vs ≥ 0.462 ng/mL). When any analysis cell had fewer than five cases, the Fisher’s exact test was used. The statistical difference between groups A and B in each clinicopathological category was determined by the Student’s t test (two-tailed) or analysis of variance. Survival curves were plotted using the Kaplan-Meier method. The statistical difference in survival between the different groups was compared using the log-rank test. Survival correlation with the prognostic factors was further investigated by multivariate analysis using the Cox proportional hazards model with backward stepwise likelihood ratio. RESULTS: ESCC patients tended to have significantly higher serum H2 RLN concentrations (0.48 ± 0.17 ng/ mL, n=141) compared with the healthy control group (0.342 ± 0.12 ng/mL, n=112). There was a significant difference between patients with lymph node involvement (0.74 ± 0.15 ng/mL, n=90), distant metastasis (0.90 ± 0.19 ng/mL, n=32) and those without lymph node involvement (0.45 ± 0.12 ng/mL, n=51), and distant metastasis (0.43 ± 0.14 ng/mL, n=109), respectively (P < 0.01). Patients with high H2 RLN levels (≥ 0.462 ng/mL) had a poorer prognosis than patients with low serum H2 RLN levels (< 0.462 ng/mL; P=0.0056). The H2 RLN level was also correlated with survival and     

mir-183在食管鳞癌中的表达水平及其与临床病理、预后的关系

目的:探讨原发性食管鳞癌(esophageal squamous cell carcinoma,ESCC)组织中mir-183的表达状况及其与食管鳞癌临床病理特征之间的关系.方法:应用实时RT-PCR方法及2-△△CT分析法分别检测ESCC患者癌组织及癌旁正常组织中mir-183的表达状况及其与临床病理特征的相关性.结果:在53例标本中,有22例(41.51%)mir-183高表达(2-6411倍),mir-183的高表达分别与患者的淋巴结转移情况及不良预后有显著统计学意义(P<0.05).结论:mir-183的高表达在ESCC的发生发展中发挥重要作用.     

新疆哈萨克族食管鳞癌中微小RNA let-7的表达及病理生物学意义

目的探讨微小RNA（miRNA）let-7在食管鳞癌细胞及组织中的表达,及let-7与食管鳞癌临床病理的关系。方法在细胞水平,运用实时荧光定量聚合酶链反应（real-time quantitative PCR,qRT-PCR）检测let-7在正常食管鳞状上皮细胞Heepic及食管鳞癌细胞CaES-17、Eca109和KYSE-150中的表达差异;在组织水平,采用qRT-PCR技术检测let-7在15对哈萨克族食管鳞癌组织和癌旁正常组织中的表达,并分析其与食管鳞癌浸润、转移等临床病理的关系。结果 qRT-PCR结果显示,let-7在3个食管鳞癌细胞系中的表达明显低于在正常食管鳞状上皮细胞Heepic中的表达,let-7在食管鳞癌组织中的表达明显低于其在癌旁正常组织中的表达,其相对表达量比值为0.79±0.56,差异有统计学意义（P〈0.05）;let-7在不同性别、分化程度、大体分型、浸润深度及淋巴结转移之间的相对表达量比值均无统计学差异（P〉0.05）。结论 Let-7在正常食管鳞状上皮细胞和食管鳞癌细胞中均有表达,但在食管鳞癌细胞中呈低表达;let-7在食管鳞癌组织和正常食管鳞状上皮组织中均有表达,但在癌组织中呈显著低表达;哈萨克族食管鳞癌患者let-7低表达与其临床病理参数之间无显著相关性;let-7对哈萨克族食管鳞癌的发生起抑制作用,是食管鳞癌的潜在分子标记物。     

Expression of thymidylate synthase and glutathione-s- transferase π in patients with esophageal squamous cell carcinoma

AIM： To investigate the expression of thymidylate synthase （TS） and glutathione-s-transferase π （GST-π） in esophageal squamous cell carcinoma and their association with the clinicopathologic characteristics. METHODS： Immunohistochemical methods were used to detect the expression of TS and GST-π in surgically resected formalin-fixed, paraffin-embedded esophageal squamous cell carcinoma （ESCC） tissue sections from 102 patients （median age, 58 years） and in 28 normal esophageal mucosa （NEM） samples. The relationship between TS and GST-π expression and clinicopathologic factors was examined. RESULTS： The expression of TS and GST-π was not statistically significantly associated with age of the patients, tumor size, lymph node metastasis, depth of invasion or tumor stage. TS staining was positive in 17.86% of normal esophageal mucosa and in 42.16% of ESCC samples （P 〈 0.05）. The expression level of TS was not only significantly lower in well-differentiated （21.88%） than in poorly-differentiated carcinomas （51.43%, P 〈 0.05）, but was also significantly higher in samples from male patients （46.51%） than from female patients （18.75%, P 〈 0.05）. GST-π was positively stained in 78.57% of normal esophageal mucosa and in 53.92% of ESCC samples （P 〈 0.05）. The expression level of GST-π was also significantly higher in welldifferentiated carcinomas （65.63%） than in poorly- differentiated carcinomas （35.00%, P 〈 0.05）. CONCLUSION： The expression of TS and of GST-π may be used as molecular markers for the characterization of ESCC. Poorly-differentiated cells showed increased expression of T5 and reduced expression of GST-π.     

Downregulation of microRNA-382 is associated with poor outcome of esophageal squamous cell carcinoma

AIM: To study the potential prognostic role of microRNA-382(miR-382) in esophageal squamous cell carcinoma(ESCC).METHODS: Forty six patients were divided into 2groups according to postoperative survival time:the poor outcome group(28 patients), who showed early metastasis but no recurrence, and died within 1year after surgery, 12 patients of the group received postoperative chemotherapy treatment that was given after early metastasis happening; the good outcome group(18 patients), who had no clinical metastasis and recurrence, and survived 5 years or more after surgery, all patients did not receive any postoperative treatment. Total RNA was extracted from the patients' formalin-fixed and paraffin-embedded esophageal cancer tissues. miR-382 level was evaluated using highthroughput real-time quantitative polymerase chain reaction analysis. The correlation between miR-382 level and clinicopathologic features was analyzed through COX regression model, and Kaplan-Meier analysis was used to analyze the relationship betweenmiR-382 level and patient survival time.RESULTS: miR-382 was differentially expressed in the two groups. Overall the average miR-382 level in the ESCC patients with good outcome was 9.8 ± 3.8,while miR-382 level in the ESCC patients with poor outcome was 3.0 ± 0.8. The differences of miR-382 levels between two groups were significant(P 0.05).Kaplan-Meier analysis results showed that miR-382 expression level generally had a significant reversecorrelation with ESCC patient survival time(P 0.001), in which the patients with higher expressions of miR-382 had a longer survival time either among individuals with the same tumor stage or among the overall patients.CONCLUSION: miR-382 levels are reverse-correlated with ESCC poor outcomes, suggesting that miR-382 could be a potential predictive biomarker for both prognosis and treatment of ESCC.     

Expression of midkine and its clinical significance in esophageal squamous cell carcinoma

AIM: TO investigate the expression of midkine in esophageal squamous cell carcinoma (ESCC) and analyze its relationship with clinicopathological features. METHODS: RT-PCR and immunocytochemical staining were used to detect the expression of midkine mRNA and protein in EC109 cells, respectively. Then the expression of midkine in 66 cases of ESCC samples were detected by immunohistochemistry using monoclonal antibodies against human midkine. RESULTS: Midkine was expressed in EC109 cell by RT-PCR and immunocytochemistry. The immunoreactivity was detected in 56.1% (37/66) of the ESCC samples. The expression of midkine was found in cytoplasm of tumor cells. Notably, the intensity of midkine was stronger at the area abundant in vessels and the invading border of the tumors. Midkine was more intensely expressed in well differentiated tumors (76.9%) than in moderately and poorly differentiated tumors (43.1% and 41.2%, respectively) (P<0.05). There was no statistically significant correlation between midkine expression and gender, age, clinical stage, lymph node metastasis or survival in ESCC. CONCLUSION: Midkine is overexpressed in ESCC. It may play a role in tumor angiogenesis and invasion. The expression of midkine is correlated with tumor cell differentiation in ESCC. The more poorly tumor cells differentiate, the weaker midkine expresses.     

Methylation of TIMP3 in esophageal squamous cell carcinoma

AIM： To measure the frequency of DNA methylation of the tissue inhibitor of metalloproteinase 3 （TIMP3） promoter and relate this to any change of gene expression in esophageal squamous cell carcinoma in patients from a region of high incidence in China. METHODS： Cancer cell lines were treated with or without the demethylating reagent 5-aza-2′-deoxycytidine. Methylation of the TIMP3 promoter was assessed in three regions by melt curve analysis and its expression was assessed by real-time RT-PCR. Tumors and proximal resection margins were obtained from 64 patients with esophageal squamous cell carcinoma from a region of high incidence in China. Methylation was assessed by melt curve analysis and expression by immunohistochemistry.
RESULTS： Methylation in one of the three promoter regions assessed correlated with gene silencing in esophageal cell lines. A degree of methylation of TIMP3 was found in only four esophageal squamous cell carcinomas, and partial loss of TIMP3 protein expression in just one.
CONCLUSION： Methylation and loss of expression of TIMP3 occurs infrequently in esophageal squamous cell carcinoma in a region of high incidence in China.     

Epstein-Barr virus association is rare in esophageal squamous cell carcinoma

Background. A critical role of Epstein-Barr virus (EBV) in carcinogenesis of nasopharyngeal squamous cell carcinoma and gastric adenocarcinoma is strongly suspected. We analyzed the possible EBV association for Japanese squamous cell carcinoma (SCC)-dominant esophageal cancer cases. Methods. We retrospectively screened 36 surgically resected esophageal cancer lesions from 36 patients maily with SCC using in situ hybridization (ISH) for EBV-encoded small RNA1 (EBER-1). EBV DNA analysis using real-time quantitative polymerase chain reaction (Q-PCR) was performed for three recent cases. Results. We found no EBER-1-positive cancer cell in any tested esophageal cancer lesion. There were many EBER-1-positive tumor-infiltrating lymphocytes in the basaloid SCC lesion and a small number of positive lymphocytes in the other five advanced SCC lesions (14.7% of SCC). One SCC lesion with a highcopy number of EBV DNA had EBER-1-positive lymphocytes. Conclusions. EBV is rarely associated with esophageal SCC, and may appear through tumor-infiltrating lymphocytes in some advanced lesions.     

N-cadherin knock-down decreases invasiveness of esophageal squamous cell carcinoma in vitro

AIM： To examine the expressions of N-cadherin and E-cadherin in specimens of 62 normal esophageal epithela, 31 adjacent atypical hyperplastic epithelia and 62 esophageal squamous cell carcinomas （ESCCs）, and to investigate the roles of N-cadherin in the invasiveness of ESCC cell line EC9706 transfected by N-cadherin shRNA.
METHODS： PV immunohistochemistry was used to detect the expression pattern of N-cadherin and E-cadherin in specimens of 62 normal esophageal epithelia, 31 adjacent atypical hyperplastic epithelia and 62 ESCCs. The invasiveness of ESCC line EC9706 was determined by transwell assay after EC9706 was transfected by N-cadherin shRNA.
RESULTS： The positive rotes of N-cadherin decreased in the carcinoma, adjacent atypical hyperplastic and normal esophageal tissues （75.8%, 61.3% and 29.0%, P 〈 0.05）, respectively, while those of E-cadherin increased （40.3%, 71.0% and 95.2%, P 〈 0.05）. The increased expression of N-cadherin and decreased expression of E-cadherin were related to invasion, differentiation, and lymph node metastasis （P 〈 0.05）. The expression level of N-cadherin decreased in the N-cadherin knocked down cells, and the invasiveness of those cells decreased significantly as well. The number of cells which crossed the basement membrane filter decreased from 123.40 ± 8.23 to 49.60 ±6.80 （P 〈 0.05）.
CONCLUSION： E-cadherin and N-cadherin expression is correlated with the invasion and aggravation of ESCC. The down-regulation of N-cadherin lowers the invasiveness of EC9706 cell line.     

基质金属蛋白酶-9和组织金属蛋白酶抑制剂-1在食管鳞癌中的表达

目的探讨基质金属蛋白酶-9（MMP-9）和组织金属蛋白酶抑制剂-1（TIMP-1）在食管鳞癌中的表达及其临床意义。方法用免疫组化和Western blot法分别检测41例食管鳞癌患者的癌及相应正常组织中MMP-9和TIMP-1的表达变化。结果食管鳞癌组织中MMP-9阳性表达率与食管癌淋巴结及静脉转移有关；MMP-9的阳性表达率与表达量均显著高于TIMP-1；MMP-9和TIMP-1的表达呈负相关。结论MMP-9与食管鳞癌的侵袭转移有关，其机制可能与食管鳞癌组织中的MMP-9／TIMP-1平衡失调有关；MMP-9与TIMP-1联合检测有助于食管鳞癌生物学行为的判断。     

Syk基因在食管鳞癌组织中的表达及临床意义

目的:分析食管鳞癌组织及其对应癌旁组织中Syk蛋白和mRNA的表达,探讨其与食管鳞癌临床恶性生物学行为的关系.方法:采用免疫组织化学法检测48例食管鳞癌组织及其对应癌旁组织中Syk蛋白的表达;RT-PCR法检测143例食管鳞癌组织及其对应癌旁组织中PSyk mRNA的表达,并观察其与食管鳞癌患者肿瘤大小、TNM分期、淋巴结转移的关系.结果:食管鳞癌组织Syk蛋白表达阳性率显著低于癌旁组织(16.67% vs 89.58%,P<0.05);Syk蛋白表达与肿瘤删分期相关(X<'2>=6.713,P<0.05):Syk在淋巴结转移组中的阳性表达率显著低于无淋巴结转移组(3.03% vs 29.41%,P<0.05);Syk表达与肿瘤大小无关(X<'2>=0.017,P>0.05).食管鳞癌组织中Syk mRNA的表达量明显低于其在对应癌旁组织中的表达量(t=-11.27,P<0.05).结论:Syk蛋白和mRNA在食管鳞癌中表达缺失与其发生及转移倾向相关,Syk可能是食管鳞癌的肿瘤抑制基因,可能作为分子标记而用于食管鳞癌的早期诊治.     

Methylenetetrahydrofolate reductase C677T polymorphism and predisposition towards esophageal squamous cell carcinoma in a German Caucasian and a northern Chinese population

Purpose Folate deficiency is considered to increase the risk of developing esophageal cancer. Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme involved in folate metabolism. A single C T substitution at nucleotide 677 of the MTHFR cDNA influences enzyme activity. The purpose of this study is to compare the association of the MTHFR C677T polymorphism with susceptibility to esophageal squamous cell carcinoma (ESCC).Methods Using real-time PCR and melting curve analysis, the MTHFR C677T genotypes were determined in 430 patients with ESCC (241 German Caucasians and 189 northern Chinese) and 397 unrelated healthy controls (256 German Caucasians and 141 northern Chinese).Results A significant difference in MTHFR C677T genotype distribution was observed between German Caucasian controls (C/C, 41.8%, C/T, 44.9%, T/T, 13.3%) and northern Chinese controls (C/C, 17.7%, C/T, 38.3%, T/T, 44.0%) (²=52.19, P<0.001). The distribution of the MTHFR C677T genotypes among German ESCC patients (C/C, 39.0%, C/T, 48.1%, T/T, 12.9%) was not significantly different from that among healthy controls (²=0.531, P=0.767). In contrast, the frequency of the C/C genotype among Chinese ESCC patients (8.5%) was significantly lower than among Chinese healthy controls (17.7%) (²=6.37, P=0.012). The C/C genotype was correlated with a significantly reduced risk for the development of ESCC as compared to the combination of C/T and T/T genotypes (adjusted OR=0.38, 95% CI=0.16–0.88).Conclusions Our results suggest that, in contrast to German Caucasians, the MTHFR 677CC homozygous wild-type plays a protective role in the development of ESCC in the northern Chinese population.Abbreviations CI Confidence interval - ESCC Esophageal squamous cell carcinoma - MTHFR Methylenetetrahydrofolate reductase - OR Odds ratio - PCR Polymerase chain reaction     

Functional studies of a novel oncogene TGM3 in human esophageal squamous cell carcinoma

AIM: To investigate the role of transglutaminase 3 (TGM3) gene in human esophageal squamous cell carcinoma (ESCC), and analyze its relationship with clinicopathological parameters. METHODS: Gene expression of TGM3 in fresh esophageal cancer tissues and their corresponding normal mucosas was detected immunohistochemically(IHC) by means of tissue microarray(TMA). Its correlation with clinical characteristics was evaluated and analyzed by univariate analysis. All statistical analyses were performed by SPSS version 10.0. RESULTS: Expression rate of TGM3 was reduced to 81.8% in ESCC. Expression of TGM3 was significantly inversely correlated with histological grade of esophageal carcinoma (in gradeⅠ,ⅡandⅢtumors, the reduced expression was 4/7,57/71, and 20/21, respectively, P < 0.05), whereas it had no obvious correlations with lymph node metastasis and depth of invasion. CONCLUSION: Reduced expression of TGM3 may play an important role in esophageal carcinogenesis.