食管鳞癌染色体及基因组DNA畸变研究进展

食管鳞癌是我国常见的消化道恶性肿瘤, 进展快且预后差。由于早期一般无明显症状, 临床确诊的食管鳞癌大多已发展到了中晚期, 治愈难度较大。越来越多的证据表明, 在食管鳞癌发生发展过程中, 染色体及基因组DNA畸变均是最常见的遗传学改变。文章就食管鳞癌染色体及基因组水平异常的研究进展作一综述。     

《自然》:科学家发现食管鳞癌相关基因突变

<正>科研人员通过高通量测序、比较基因组杂交芯片分析及生物学功能研究,全面系统揭示了食管鳞癌的遗传突变背景,发现了一批与食管鳞癌发生发展进程和临床预后相关的基因,为了解食管鳞癌的发病机理,寻找食管鳞癌诊断的分子标志物,确定研发临床治疗的药物靶点以及制定有效的治疗方案提供了理论和实验依据。3月17日,研究成果于国际学术权威杂志《自然》在线发表。食管癌是人类常见的恶性肿瘤之一,在我国和全球所有恶性肿瘤死亡率中分别排在第四位和第六位。目前,食管癌发生和发展的机制尚不明了,治疗缺乏特异性的分子靶点和有效的治疗药物。科研人员对我国食管鳞癌患者样本进行了全基因组测序、全外显子组测序和比较基因组杂交     

血清miRNAs作为食管鳞癌新的生物标志物

食管鳞癌是我国最常见的恶性肿瘤之一。由于缺乏有效的早期诊断方法,大多数食管鳞癌患者在确诊时已到中晚期并预后不良。MicroRNAs(miRNAs)是一类可通过抑制其特异性靶基因表达从而调控食管鳞癌发生发展的非编码内源性小RNA。相比于传统的生物标志物(例如mRNA和蛋白质),miRNAs更加稳定并易于筛选及精确地定量分析,从而成为理想的新一代癌症早期诊断和预后评估的生物标志物。近来的研究结果显示,食管鳞癌病人血清中的一些miRNAs表达水平的变化与病情诊断及预后的结果显著相关。文章综述了食管鳞癌病人血清中miRNAs的变化规律,讨论了检测这些miRNAs的表达水平变化作为一种新的方法应用于食管鳞癌的早期诊断和预后评估的可能性。值得注意的是,不同的血清miRNAs的检测方法所产生的结果是不完全一致的,文章还对这些差异产生的原因进行了讨论。     

MRP2 蛋白在食管鳞癌中的表达及其与化疗耐药的相关性

目的:探讨MRP2蛋白在食管鳞癌组织中的表达及其与食管癌化疗耐药的关系。方法:收集原发性食管鳞癌手术标本70例,采用免疫组织化学Envision法检测食管鳞癌组织及其癌旁组织中MRP2蛋白的表达情况,并采用MTT法检测食管鳞癌组织对临床常用化疗药物的敏感性,分析其表达与食管癌化疗耐药的关系。结果:70例食管鳞癌组织及其癌旁正常组织中的阳性表达率分别为58.6%及5.0%。MRP2蛋白在食管鳞癌组织中的阳性表达率明显高于癌旁正常食管组织(P<0.01)。食管鳞癌组织对环磷酰胺、5-氟尿嘧啶、吉西他滨、顺铂、卡铂、阿霉素、长春瑞滨、羟喜树碱等化疗药物的敏感性与其相应癌组织中MRP2表达明显相关(P<0.01)。结论:MRP2的表达与食管鳞癌对多种化疗药物耐药有较好的相关性,推测食管鳞癌组织中MRP2的高表达可能对化疗耐药性的发生发展具有促进作用。     

LncRNA MEG3抑制食管鳞癌组织及KYSE30细胞

本研究通过检测长链非编码RNA(lncRNA)MEG3在食管鳞癌组织及人KYSE30细胞中的表达情况,分析lncRNA MEG3在食管鳞癌发生发展中的作用。我们采集食管鳞癌手术肿瘤组织54例作为观察组,癌旁正常组织54例作为对照组,采用RT-PCR技术检测MEG3表达水平。进行MEG30过表达及抑制实验,检测KYSE30细胞增殖及侵袭活性的变化。研究结果显示观察组食管鳞癌组织中lncRNA MEG3表达低于对照组的正常组织,差异具有统计学意义(p0.05);lncRNA MEG3过表达后KYSE30细胞增殖及侵袭能力减弱,lncRNA MEG3表达抑制后,KYSE30细胞增殖和侵袭能力增强。结果表明,lncRNA MEG3对食管鳞癌的发生发展有抑制作用。     

食管鳞癌组织中TNFAIP3的表达及其预后意义

目的研究肿瘤坏死因子α诱导蛋白3(tumor necrosis factorα-induced protein 3, TNFAIP3)在食管鳞癌中表达的临床病理意义及其对鳞癌预后的判断价值。方法免疫组织化学法检测100例随访食管鳞癌及80例癌旁组织中TNFAIP3蛋白的表达,并分析了其与食管鳞癌临床病理特征及预后的关系。结果 TNFAIP3在食管鳞癌中高表达阳性率为47.0%(47/100),显著高于癌旁组织中的高表达率15.0%(12/80),且与肿瘤的分化程度有关;生存分析显示高表达TNFAIP3的患者预后不良,单因素生存分析显示TNFAIP3的表达、肿瘤的浸润深度、淋巴结转移、临床分期与食管鳞癌预后密切相关,多因素生存分析显示TNFAIP3的表达、肿瘤的浸润深度、淋巴结转移、临床分期为食管鳞癌的独立预后预测因子。结论表达增高的TNFAIP3可能参与了食管鳞癌的发生发展,TNFAIP3的高表达可能成为食管鳞癌的独立预后因子。     

食管鳞癌上皮间质转化与肿瘤侵袭转移的研究进展

食管癌是全球常见的恶性肿瘤之一,在中国主要以鳞癌为主。虽然近年来食管鳞癌的早期诊断、手术治疗以及局部和全身的辅助治疗等方面的研究进展显著,但食管鳞癌的发病率和死亡率仍然居高不下,其中肿瘤的侵袭和转移是导致患者死亡的最主要原因。肿瘤细胞的侵袭和转移能力主要通过上皮间质转化(epithelial-mesenchymal transition,EMT)过程来获得。EMT是一个动态、多步骤和高度复杂的过程,其中涉及到肿瘤细胞基因、表观遗传学等的改变,这些改变与肿瘤的侵袭和转移密切相关。本文综述了食管鳞癌EMT与肿瘤侵袭转移的研究进展,旨在为食管鳞癌的诊断治疗研究积累基础理论资料。     

DNA修复酶MYH在食管鳞癌的表达和临床意义

目的:探讨食管鳞癌组织MYH的表达与8-oxoG氧化损伤和临床病理特征之间的关系。方法:用免疫组化染色和Western blot实验,比较食管鳞癌组织和癌旁组织MYH表达的高低;用免疫组化染色比较食管鳞癌组织和癌旁组织8-oxoG氧化损伤程度的高低。统计分析食管鳞癌组织MYH表达的高低与患者临床和病理特征的关系,采用x2检验。结果:食管鳞癌组织MYH蛋白表达低于其癌旁组织,食管鳞癌组织8-oxoG氧化损伤程度高于其癌旁组织。食管鳞癌组织MYH蛋白低表达,与该组织8-oxoG氧化损伤程度、浸润深度、静脉侵犯、TNM分期、淋巴结转移有关,与年龄、性别、病理分化程度无关。结论:食管鳞癌组织MYH蛋白低表达,可能与食管鳞癌的发展有关,后常规对患者食管癌标本做MYH表达的检测,可指导食管鳞癌术后化学治疗方案的制定。     

食管鳞癌血管生成拟态的组织学和细胞学研究

目的:探讨血管生成拟态(vasculogenic mimicry,VM)与食管鳞癌临床病理特征的关系及其对患者预后的影响,并分析食管癌血管生成拟态的形成机制。方法:收集57例食管鳞癌石蜡包埋样本,进行过碘酸雪夫氏(PAS)及CD34免疫组织化学双重染色,结合HE染色,观察食管鳞癌血管生成拟态的发生情况。对患者临床病理和预后信息进行单因素分析,Kaplan-Meier生存比较和Cox风险模型分析。通过食管鳞癌细胞株Eca-109三维培养建立,观察RNAi沉默VE-cadherin对食管鳞癌Eca109血管生成拟态形成的影响。结果:食管鳞癌中VM表达的阳性率为54.3%,显著高于正常食管黏膜组织;VM在病理分型为低分化食管鳞癌的阳性表达率为78.9%,显著高于中高分化组(P0.05);III-Ⅳ期食管鳞癌患者VM阳性率显著高于Ⅰ-Ⅱ期食管鳞癌患者(P0.05);有淋巴结转移的食管鳞癌者VM阳性率明显高于无淋巴结转移者(P0.05)。单因素分析结果显示食管鳞癌VM的发生率与肿瘤的分化程度、TNM分期和淋巴转移显著相关。Kaplan-Meier生存分析显示有VM组食管鳞癌患者的生存期明显短于无VM组(P0.05);Cox分析显示VM是影响食管鳞癌患者预后的独立危险因素(RF=0.67)。三维培养结果显示Eca-109细胞在基质胶上形成典型的血管网状样结构,VE-cadherin-siRNA可有效抑制VE-cadherin在Eca109的表达,抑制体外培养的Eca109细胞VM的形成。结论:血管生成拟态是食管鳞癌一种独特的血液供应模式,与食管鳞癌的分化程度、TNM分期、淋巴转移密切相关,是食管鳞癌患者术后生存期的独立危险因素。     

食管鳞癌中P-P38和VEGF蛋白的表达及意义

目的:探讨P-P38蛋白和VEGF蛋白在食管鳞癌组织中可能的作用.方法:采用微波EliVisionTM免疫组织化学法检测60例食管鳞癌组织中P-P38蛋白和VEGF蛋白的表达情况,并与40例正常食管粘膜组织进行比较.结果:食管鳞癌组织中P-P38和VEGF分别表达定位于细胞核和细胞浆,其表达明显高于正常食管粘膜组织(P＜0.05).两者的表达与淋巴结转移、分化程度及病理分期有关(P＜0.05);两者之间表达呈正相关(P＜0.05).结论:P-P38和VEGF在食管鳞癌呈高表达,且在食管鳞癌的发生、发展和转移过程中起到一定的作用.     

Serum metabolomics for early diagnosis of esophageal squamous cell carcinoma by UHPLC-QTOF/MS

Introduction

Previous metabolomics studies have revealed perturbed metabolic signatures in esophageal squamous cell carcinoma (ESCC) patients, however, most of these studies included mainly late-staged ESCC patients due to the difficulties of collecting the early-staged samples from asymptotic ESCC subjects.

Objectives

This study aims to explore the early-staged ESCC metabolic signatures and potential of serum metabolomics to diagnose ESCC at early stages.

Methods

Serum samples of 97 ESCC patients (stage 0, 39 cases; stage I, 17 cases; stage II, 11 cases, stage III, 30 cases) and 105 healthy controls (HC) were enrolled and randomly separated into training data (77 ESCCs, 84 HCs) and validation data (20 ESCCs, 21 HCs). Untargeted metabolomics was performed to identify ESCC-related metabolic signatures.

Results

The global metabolomics profiles could clearly distinguish ESCC from HC in training data. 16 ascertained metabolites were found to be disturbed in the metabolic pathways characterized by dysregulated fatty acid biosynthesis, glycerophospholipid metabolism, choline metabolism in cancer and linoleic acid metabolism. The AUC value in validation data was 0.895, with sensitivity 85.0 % and specificity 90.5 %. Good diagnostic performances were also achieved for early stage ESCC, with the values of area under the curve (AUC) 0.881 for the ESCC patients in both stage 0 and I–II. In addition, six metabolites were found to discriminate ESCC stages. Among them, three biomarkers, dodecanoic acid, LysoPA(18:1), and LysoPC(14:0), exhibited clear trend for ESCC progression.

Conclusion

These findings suggest serum metabolomics, performed in a minimally noninvasive and convenient manner, may possess great potential for early diagnosis of ESCC patients.

     

Comprehensive bioinformatics analysis identifies lncRNA HCG22 as a migration inhibitor in esophageal squamous cell carcinoma

Esophageal cancer is one of the most lethal malignancies worldwide, and esophageal squamous cell carcinoma (ESCC) is the dominant histological type. However, the long noncoding RNA (lncRNA) alterations in ESCC have not been elucidated to date. In this study, reliable databases from Gene Expression Omnibus (GEO), which analyzed lncRNA expression in ESCC tumor tissues and adjacent normal tissues were searched, and common differentially expressed lncRNAs and genes were analyzed. Next, cis- trans analysis was performed to predict the underlying relationships between altered lncRNAs and mRNAs, and the lncRNA-mRNA regulatory network was established. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of altered lncRNA-related genes were performed. The promising lncRNA HCG22 was validated by quantitative polymerase chain reaction (qPCR), and clinicopathological data were collected to identify the relationship between lncRNA HCG22 expression level and clinical features. Finally, Transwell assays were performed to explore the biological functions of lncRNA HCG22 in ESCC cells. Two hundred forty-one lncRNAs and 835 mRNAs were observed to be remarkably altered between ESCC tumor tissues and adjacent normal tissues. The lncRNA-mRNA regulatory network showed the coexpression association between lncRNA HCG22 and SPINK7 and ADAMTS12. GO and KEGG analyses showed that HCG22 and ADAMTS12 had potential biological functions in the cell migration of ESCC. The downregulation of lncRNA HCG22 in ESCC tumor tissues was validated by qPCR, and the clinicopathological data showed a noticeable correlation between lncRNA HCG22 expression level and the ESCC differentiational degree and clinical TNM stage. Kaplan-Meier analysis showed that patients with ESCC having low lncRNA HCG22 expression in ESCC tissues had considerably shorter overall survival compared with patients with ESCC having high lncRNA HCG22 expression. Following Transwell assays confirmed the migratory role of lncRNA HCG22 in ESCC cells. In conclusion, lncRNA HCG22 was downregulated in ESCC tissues and can be a migration inhibitor of ESCC cells, and SPINK7 and ADAMTS12 are promising to be the regulatory targets of lncRNA HCG22.     

H2-Calponin 在食管鳞癌中的表达及其与食管磷癌预后的关系

目的:探讨H_2-Calponin在食管磷癌组织中的表达及其与食管磷癌患者临床病理特征及预后的相关性。方法:利用免疫组织化学染色技术检测73例食管磷癌及相应癌旁组织中H_2-Calponin的表达情况,并分析其与食管磷癌患者临床病理参数及预后的关系。结果:H_2-calponin在食管磷癌组织中的阳性表达率为95.8%(70/73),显著高于癌旁组织(35.8%,24/67),差异具有统计学意义(p0.0001)。H_2-calponin的表达与食管癌患者的性别、年龄、肿瘤大小、浸润深度及淋巴结转移均无显著相关性,但与AJCC分期显著相关(P0.05)。H_2-calponin的表达水平影响食管癌患者预后及生存期。结论:H_2-Calponin在食管磷癌组织中呈高表达并可预示食管磷癌预后不良。     

A six-microRNA risk score model predicts prognosis in esophageal squamous cell carcinoma

Esophageal cancer ranks the eighth most common cancer and the sixth most common cause of cancer death worldwide. MicroRNAs (miRNAs) are small noncoding RNAs that regulate a wide variety of cancer-related cellular processes. In the current study, a series of previously published gene expression microarray data from Gene Expression Ominus and The Cancer Genome Atlas were downloaded and further divided into training, internal, and external validation sets. Least absolute shrinkage and selectionator operator Cox regression model along with 10-fold cross-validation was performed to select the miRNAs associated with the prognosis of esophageal squamous cell carcinoma (ESCC) and constructed a six-miRNA signature. Then the prediction accuracy of this signature was assessed in validation and test set using Kaplan–Meier analysis, time-dependent receiver operating characteristic (ROC) curves and dynamic area under the ROC curve. According to the result, the prediction accuracy of miRNA signature was much better than that of tumor–node–metastasis (TNM) stage in all the three sets. Stratified analysis also demonstrated that the predict ability of this signature was independent of TNM stage. Finally, function experiments including apoptosis and colony formation assay were performed to further reveal the regulatory role of miRNAs in ESCC. Our study demonstrated the promising potential application of this novel six-miRNA signature as an independent biomarker for survival prediction of ESCC patients.     

Up-regulated miR-548k promotes esophageal squamous cell carcinoma progression via targeting long noncoding RNA-LET

Dysregulated noncoding RNAs have been observed in diverse cancers. MIR458K is frequently amplified in esophageal squamous cell carcinoma (ESCC). However, the expression, clinical significances, and action mechanisms of miR-548k in ESCC are still unclear. In this study, we found that miR-548k is significantly up-regulated in ESCC tissues and cell lines. Up-regulated miR-548k expression is significantly correlated with advanced invasion depth, lymph node metastasis, advanced TNM stage, and poor overall survival. Gain-of- and loss-of-function assays demonstrated that miR-548k promotes the proliferation and migration of ESCC cells in vitro and tumor growth in vivo. Mechanistically, we found that miR-548k directly targets and represses the expression of long noncoding RNA-LET (lncRNA-LET), and further down-regulates p53 and up-regulates NF90. In addition, we found that lncRNA-LET is down-regulated and inversely correlated with miR-548k in ESCC. Down-regulated lncRNA-LET also indicated poor overall survival of ESCC patients. Functional assays demonstrated that lncRNA-LET inhibits the proliferation and migration of ESCC cells, and the effects of miR-548k on ESCC are dependent on the negative regulation of lncRNA-LET. In summary, our data revealed the critical roles of miR-548k-lncRNA-LET regulation axis in ESCC and suggested that the miR-548k-lncRNA-LET regulation axis may be promising prognostic biomarkers and therapeutic targets for ESCC.     

maspin在食管鳞状细胞癌中的表达及其与微血管密度的关系

目的研究旨在寻找能预测食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)浸润、转移及术后生存率的生物因子。方法采用免疫组织化学ElivisionTM plus法检测100例ESCC和30例正常食管组织中maspin的表达和微血管密度(microvessel density,MVD)情况。结果在正常食管组织和ESCC组织中,maspin的阳性表达率分别为100%和43.0%,差异有显著性(P<0.01);maspin的表达水平与肿瘤的分化程度、浸润深度、淋巴结转移和临床分期有关(全部P<0.01);MVD计数与肿瘤的大小、分化程度、淋巴结转移以及临床分期有关(全部P<0.01);且maspin的表达与MVD计数呈负相关(P<0.01)。结论 maspin的表达和MVD计数与ESCC组织的分化程度、转移和预后等均有关;maspin和MVD联合检测对ESCC的进展及预后判断有重要意义。     

MicroRNA-143 functions as a tumor suppressor in human esophageal squamous cell carcinoma

Esophageal cancer is one of the most common cancers worldwide with a poor prognosis. MicroRNAs(miRNAs) are a class of naturally occurring small noncoding RNAs and play an important role in cancer initiation and development. In this study, we demonstrate that the expression levels of miR-143 and miR-145 were significantly decreased in ESCC tissues in comparison with adjacent normal esophageal squamous tissues(NESTs). Furthermore, an inverse correlation between miR-143 and tumor invasion depth and lymph node metastasis was observed. The enforced expression of miR-143 induced growth suppression and apoptosis of ESCC cells. Rescue of miR-143 significantly suppressed the ESCC cells migration and invasion capabilities. Moreover, we show that functions of miR-143 in ESCC are mediated at least in part by the inhibition of extracellular signal regulated kinase-5(ERK-5) activity. These results prove that miR-143 may act as a tumor suppressor in ESCC.     

Reduced LINC00551 expression promotes proliferation and invasion of esophageal squamous cancer by increase in HSP27 phosphorylation

Esophageal squamous cell carcinoma (ESCC) is one of the deadliest cancers, and long noncoding RNAs (lncRNAs) regulate gene expression or activities. This study investigated the role of lncRNA LINC00551 in ESCC development and progression. Three paired ESCC and normal tissues were subjected to next‐generation sequencing and we identified 82 upregulated and 60 downregulated lncRNAs, including LINC00551, which was confirmed to markedly downregulated in 78 ESCC tissues and in the Gene Expression Profiling Interactive Analysis data set. Downregulated LINC00551 expression was associated with lymph node metastasis, advanced TNM stage, and tumor size. Moreover, downregulated LINC00551 expression was also associated with poor progression‐free survival and overall survival of ESCC patients. In vitro and in vivo, LINC00551 overexpression inhibited ESCC cell proliferation and invasion, whereas knockdown of LINC00551 expression promoted ESCC cell proliferation and invasion. RNA pull‐down and mass spectrometry assays identified the potential LINC00551 binding proteins, and HSP27 was a promising LINC00551 targeting proteins after RNA immunoprecipitation assay. At the protein level, LINC00551 bound to and decreased HSP27 phosphorylation, and in turn, downregulated ESCC cell proliferation and invasion. The current study demonstrated the functional significance of LINC00551 in ESCC development, progression, and prognosis. Further study will assess LINC00551 as a novel prognostic marker or therapeutic target for ESCC.     

Overexpression of phosphatase regenerating liver 3 in oesophageal squamous cell carcinoma associated with metastasis and its comparison with phosphatase regenerating liver 1

Expression of PRL3 (phosphatase of regenerating liver 3) protein was examined with immunohistochemistry in 60 cases of ESCC (oesophageal squamous cell carcinoma) with matched lymph node metastasis (n = 40) and 6 cases of oesophageal adenocarcinoma. Its associations with PRL1 and clinicopathological parameters were analysed. The results showed the frequency of PRL3 protein expression was significantly higher in ESCC (39/60, 65%) than in normal oesophageal mucosa (0/20, P < 0.001); higher in ESCC with lymph node metastasis (30/40, 75%) than in ESCC without lymph node metastasis (9/20, P = 0.022), as well as higher in metastatic ESCC in lymph node (38/40, 95%) than in the primary ESCC (39/60, 65%, P < 0.001). PRL3 was expressed in 1 out of 6 oesophageal adenocarcinomas, but showed no nuclear staining of PRL1. Expression of PRL3 protein was positively associated with the grade and partially with the stage of ESCC. These results suggest that expression of PRL3 protein may be involved in the metastasis of ESCC and serve as a biomarker for prediction of ESCC metastasis.