BACKGROUND & AIMS: :Esophageal squamous cell carcinoma is the sixth most lethal cancer worldwide and the fourth most lethal cancer in China. Tissue-specific transplantation antigen P35B codifies the enzyme GDP-d-mannose-4,6-dehydratase, which participates in the biosynthesis of GDP-l-fucose. GDP-l-fucose is an important substrate involved in the biosynthesis of many glycoproteins. Cancer cells are often accompanied by the changes in glycoprotein structure, which affects the adhesion, invasion, and metastasis of cells. It is not clear whether tissue-specific transplantation antigen P35B has any effect on the development of esophageal squamous cell carcinoma. We used an immunohistochemical method to assess the expression of tissue-specific transplantation antigen P35B in 104 esophageal squamous cell carcinoma samples. The results showed tissue-specific transplantation antigen P35B expression was associated with some clinical features in patients, such as age ( P = .017), clinical stage ( P = .010), and lymph node metastasis ( P = .043). Kaplan-Meier analysis and log-rank test showed that patients with esophageal squamous cell carcinoma having high tissue-specific transplantation antigen P35B expression had a worse prognosis compared to the patients with low expression ( P = .048). Multivariate Cox proportional hazards regression model showed that high expression of tissue-specific transplantation antigen P35B could predict poor prognosis for patients with esophageal squamous cell carcinoma independently. In conclusion, abnormal fucosylation might participate in the progress of esophageal squamous cell carcinoma and tissue-specific transplantation antigen P35B may serve as a novel biomarker for prognosis of patients with esophageal squamous cell carcinoma.

背景与目标： ：食道鳞状细胞癌是全球第六大致死性癌症，也是中国第四大致死性癌症。组织特异性移植抗原P35B编码了GDP-d-甘露糖-4,6-脱水酶，该酶参与GDP-1-岩藻糖的生物合成。 GDP-1-岩藻糖是参与许多糖蛋白生物合成的重要底物。癌细胞通常伴随着糖蛋白结构的变化，从而影响细胞的粘附，侵袭和转移。尚不清楚组织特异性移植抗原P35B是否对食道鳞状细胞癌的发展有任何影响。我们使用免疫组织化学方法评估了104例食管鳞状细胞癌样品中组织特异性移植抗原P35B的表达。结果显示组织特异性移植抗原P35B的表达与患者的某些临床特征有关，例如年龄（P = .017），临床分期（P = .010）和淋巴结转移（P = .043）。 Kaplan-Meier分析和对数秩检验表明，组织特异性移植抗原P35B表达高的食管鳞状细胞癌患者与低表达的患者相比，预后较差（P = .048）。多元Cox比例风险回归模型显示，组织特异性移植抗原P35B的高表达可独立预测食管鳞状细胞癌患者的预后不良。结论：岩藻糖基化异常可能参与了食管鳞状细胞癌的进展，组织特异性移植抗原P35B可能成为食管鳞状细胞癌患者预后的新生物标志物。

BACKGROUND & AIMS: :Esophageal squamous cell carcinoma (ESCC) is a common cancer occurring in males and females worldwide. Accumulating evidence continues to highlight the crucial roles of long non-coding RNAs (lncRNAs) in the process of tumorigenesis. However, the regulatory mechanism of lncRNAs in ESCC remains unclear. The aim of this study is to elucidate the role of lncRNA Krüppel-like factor 3 antisense RNA 1 (KLF3-AS1) in ESCC by regulating miR-185-5p and KLF3. Initially, ESCC cell spheres with stem cell-like properties were prepared by suspension culture, and subsequently characterized by assessing colony formation ability and stem cell markers. LncRNA KLF3-AS1 was found to be poorly expressed in ESCC and could upregulate the expression of KLF3 by binding to miR-185-5p. lncRNA KLF3-AS1 upregulation was observed to inhibit miR-185-5p, thereby contributing to decreased expression of SOX2 and Oct4 (octamer-binding transcription factor 4). Furthermore, enhancement of lncRNA KLF3-AS1 resulted in reduced colony formation ability, cell invasion and migration, and tumor volume in vivo while promoting cell apoptosis in ESCC through downregulation of miR-185-5p. Collectively, this study indicated that lncRNA KLF3-AS1 inhibited ESCC cell invasion and migration by impairing miR-185-5p-mediated inhibition of KLF3, highlighting a promising novel potential target for ESCC treatment.

背景与目标： ：食管鳞状细胞癌（ESCC）是一种常见的癌症，在世界各地的男性和女性中都有发生。越来越多的证据继续突出了长非编码RNA（lncRNA）在肿瘤发生过程中的关键作用。但是，尚不清楚lncRNA在ESCC中的调控机制。本研究的目的是通过调节miR-185-5p和KLF3阐明lncRNAKrüppel样因子3反义RNA 1（KLF3-AS1）在ESCC中的作用。最初，通过悬浮培养制备具有干细胞样特性的ESCC细胞球，然后通过评估菌落形成能力和干细胞标志物来表征。发现LncRNA KLF3-AS1在ESCC中表达较差，并可能通过与miR-185-5p结合而上调KLF3的表达。观察到lncRNA KLF3-AS1上调抑制了miR-185-5p，从而导致SOX2和Oct4（八聚体结合转录因子4）的表达降低。此外，lncRNA KLF3-AS1的增强导致体内集落形成能力降低，细胞侵袭和迁移以及体内肿瘤体积减少，同时通过下调miR-185-5p促进ESCC细胞凋亡。总体而言，这项研究表明，lncRNA KLF3-AS1通过削弱miR-185-5p介导的KLF3抑制作用来抑制ESCC细胞的侵袭和迁移，从而突出了有望用于ESCC治疗的新型潜在靶标。

BACKGROUND & AIMS: PURPOSE:To examine the global gene expression of cancer-related genes in esophageal squamous cell carcinoma (ESCC) through the use of Atlas Human Cancer Array membranes printed with 588 well-characterized human genes involved in cancer and tumor biology. EXPERIMENTAL DESIGN:Two human ESCC cell lines (HKESC-1 and HKESC-2) and one morphologically normal esophageal epithelium tissue specimen from the patient of which the HKESC-2 was derived were screened in parallel using cDNA expression arrays. The array results were additionally validated using semiquantitative PCR. The overexpression of oncogene MET was studied more extensively for its protein expression by immunohistochemistry in the two ESCC cell lines and their corresponding primary tissues and 61 primary ESCC resected specimens. Sixteen of these 61 ESCC cases also had available the corresponding morphologically normal esophageal epithelium tissues and were also analyzed for MET expression. The clinicopathological features associated with overexpression of the MET gene were also correlated. RESULTS:The results of cDNA arrays showed that 13 cancer-related genes were up-regulated > or =2-fold (CDC25B, cyclin D1, PCNA, MET, Jagged 2, Integrin alpha3, Integrin alpha6, Integrin beta4, Caveolin-2, Caveolin-1, MMP13, MMP14, and BIGH3) and 5 genes were down-regulated > or =2-fold (CK4, Bad, IGFBP2, CSPCP, and IL-1RA) in both ESCC cell lines at the mRNA level. Semiquantitative RT-PCR analysis of 9 of these differentially expressed genes, including the MET gene, gave results consistent with cDNA array findings. The immunostaining results of the expression of MET gene showed that MET was overexpressed in both ESCC cell lines and their corresponding primary tumors at the protein level, validating the cDNA arrays findings. The results of the clinical specimens showed that the MET gene was overexpressed in ESCC compared with normal esophageal epithelium in 56 of 61 cases (92%). Moreover, the overexpression of MET protein was more often seen in well/moderately differentiated than in poorly differentiated ESCC. CONCLUSIONS:Multiple cancer-related genes are differentially expressed in ESCC, the oncogene MET is overexpressed in ESCC compared with normal esophageal epithelium, and its protein overexpression correlates with tumor differentiation in ESCC.

背景与目标： 目的：通过使用Atlas Human Cancer Array膜印制了588个与癌症和肿瘤生物学有关的人类特征基因，研究了食管鳞状细胞癌（ESCC）中与癌症相关的基因的全球基因表达。
实验设计：使用cDNA表达阵列平行筛选了两个人ESCC细胞系（HKESC-1和HKESC-2）和一个形态正常的食管上皮组织标本，该标本来自于该患者的HKESC-2。使用半定量PCR进一步验证了阵列结果。通过免疫组织化学在两个ESCC细胞系及其相应的主要组织和61个主要ESCC切除的标本中对癌基因MET的过表达进行了更广泛的研究，研究了其蛋白表达。在这61例ESCC病例中，有16例具有相应的形态正常的食管上皮组织，并进行了MET表达的分析。与MET基因过表达相关的临床病理特征也相关。
结果：cDNA阵列结果显示13个与癌症相关的基因上调>或= 2倍（CDC25B，cyclin D1，PCNA，MET，锯齿状2，整合素alpha3，整合素alpha6，整合素beta4，Caveolin-2，在两种ESCC细胞系中，Caveolin-1，MMP13，MMP14和BIGH3）和5个基因在mRNA水平下调了>或= 2倍（CK4，Bad，IGFBP2，CSPCP和IL-1RA）。对这些差异表达基因中的9个（包括MET基因）进行的半定量RT-PCR分析得出的结果与cDNA阵列发现相符。 MET基因表达的免疫染色结果表明，MET在ESCC细胞系及其相应的原发性肿瘤中均在蛋白质水平上过表达，证实了cDNA阵列的发现。临床标本的结果显示，在61例患者中有56例（92％）与正常食管上皮相比，MET基因在ESCC中过表达。此外，在分化良好/中等分化的胚胎干细胞中，与分化程度低的ESCC相比，经常发现MET蛋白的过表达。
结论：与正常食管上皮相比，多种癌相关基因在食管鳞癌中差异表达，癌基因MET在食管鳞癌中过表达，其蛋白过表达与食管鳞癌的分化有关。

BACKGROUND & AIMS: ETHNOPHAMACOLOGICAL RELEVANCE:Ganoderma lucidum has been used as a medicinal mushroom for more than 2000 years in China. Ganoderic acid D (GAD) as a representative active triterpenoid from Ganoderma lucidum is known to possess anticancer activity. However, the mechanism involved in its anticancer cell process is still largely elusive. AIM OF THE STUDY:Our study aimed to investigate the anticancer effects of GAD on the esophageal squamous cell carcinoma (ESCC) cells and the underlying mechanisms at the cell level. MATERIALS AND METHODS:EC9706 and Eca109 cells were treated with GAD (0, 10, 20, 40 μM) for 24 h. The cell viability, cell cycle, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), apoptosis rate, caspase-3 activity, autophagic flux, lysosomal function were examined. Cell cycle, apoptotic, autophagy and mTOR signal pathway related proteins such as P53, Cyclin B1, CytoC, PARP, Beclin-1, P62, LC3, PI3K, AKT and mTOR were analyzed by Western blot approach. RESULTS:GAD inhibited cell proliferation and induced both apoptosis and autophagic cell death. In particular, we found that in the early stage of the autophagic process, GAD could initiate and enhance the autophagy signal while in the late stage it on the contrary could block the autophagic flux by impairing the autophagosome-lysosome fusion and inhibited the lysosomal degradation. Besides the autophagic cell death, GAD also induced the apoptosis mediated by caspase-related process in parallel. The mechanism involved for the synergistic apoptotic and autophagic cell death was also explored. We found that GAD down-regulated the expression of PI3K, AKT and mTOR phosphorylated proteins in the mTOR signaling pathway which thus led to the synergistic effect on apoptosis and autophagic cell death in the ESCC cells. CONCLUSIONS:In summary, this study has documented that GAD may inhibit cell proliferation through the mTOR pathway in ESCC cells, and induce synergistic apoptosis and autophagic cell death by disrupting the autophagic flux. This work therefore also suggests that GAD may be used as an efficient anticancer adjuvant for ESCC cancer therapy.

背景与目标： 人种药理关系：灵芝在中国已被用作药用蘑菇已有2000多年的历史了。灵芝D（GAD）作为灵芝的代表活性三萜类化合物，已知具有抗癌活性。然而，其抗癌细胞过程所涉及的机制仍然很难捉摸。
研究目的：我们的研究旨在探讨GAD对食管鳞状细胞癌（ESCC）细胞的抗癌作用及其在细胞水平上的潜在机制。
材料与方法：将EC9706和Eca109细胞用GAD（0、10、20、40μM）处理24小时。检查细胞活力，细胞周期，活性氧（ROS），线粒体膜电位（MMP），凋亡率，caspase-3活性，自噬通量，溶酶体功能。通过蛋白质印迹法分析了细胞周期，凋亡，自噬和与mTOR信号通路相关的蛋白，例如P53，Cyclin B1，CytoC，PARP，Beclin-1，P62，LC3，PI3K，AKT和mTOR。
结果：GAD抑制细胞增殖，诱导细胞凋亡和自噬细胞死亡。特别是，我们发现在自噬过程的早期，GAD可以启动并增强自噬信号，而在后期则可以通过损害自噬体-溶酶体融合来阻止自噬通量并抑制溶酶体降解。除了自噬细胞死亡，GAD还平行诱导了由caspase相关过程介导的凋亡。还探讨了协同的凋亡和自噬细胞死亡的机制。我们发现GAD下调了mTOR信号通路中PI3K，AKT和mTOR磷酸化蛋白的表达，从而导致了ESCC细胞对细胞凋亡和自噬细胞死亡的协同作用。
结论：总的来说，这项研究已经证明GAD可以通过mTOR途径抑制ESCC细胞中的细胞增殖，并通过破坏自噬通量来诱导协同凋亡和自噬细胞死亡。因此，这项工作还表明GAD可用作ESCC癌症治疗的有效抗癌佐剂。

BACKGROUND & AIMS: BACKGROUND AND AIM:The miR-196a2 gene contains a C/T polymorphism (rs11614913). Its presence could change the conformation of secondary structure of miR-196a2 RNA, and directly affect the binding to target mRNAs and the miRNA maturation process. Both of which eventually alter protein expression and contributed to cancer susceptibility. This study assessed whether the rs11614913 single nucleotide polymorphism (SNP) could affect an individual's susceptibility to esophageal squamous cell carcinomas (ESCC). METHODS:SNP rs11614913 was genotyped by polymerase chain reaction ligase detection reaction (PCR-LDR) in 597 ESCC patients and 597 control subjects. RESULTS:Overall, there were no significant differences in the frequency of the miRNA-196a2 SNP rs11614913 genotype between the ESCC cases and the controls (χ(2) = 1.395, p = 0.498). The TT genotype, CT genotype and CT/TT combined genotype (dominant model) did not modify the risk of ESCC as compared with the CC genotype. Comparisons of the TT genotype to the CT/CC combined genotype did not reveal a significant association to ESCC, too. However, further analyses revealed an increased risk of ESCC in the dominant model (OR = 1.56, 95% CI = 1.08-2.26) and the allele frequency comparison (OR = 1.31, 95% CI = 1.06-1.63) in the ≤60-year-old group. CONCLUSIONS:These results suggest that the miRNA-196a2 functional polymorphism rs11614913 might be an effective genetic marker for ESCC risk assessment in individuals younger than 60 years of age from a region of high ESCC incidence in northern China.

背景与目标： 背景与目的：miR-196a2基因具有C / T多态性（rs11614913）。它的存在可以改变miR-196a2 RNA二级结构的构象，并直接影响与靶mRNA的结合以及miRNA的成熟过程。两者最终都会改变蛋白质表达，并导致癌症易感性。这项研究评估了rs11614913单核苷酸多态性（SNP）是否会影响个体对食道鳞状细胞癌（ESCC）的敏感性。
方法：采用聚合酶链反应-连接酶检测反应（PCR-LDR）对597例ESCC患者和597例对照者进行SNP rs11614913基因分型。
结果：总体上，ESCC病例与对照组之间的miRNA-196a2 SNP rs11614913基因型频率没有显着差异（χ（2）= 1.395，p = 0.498）。与CC基因型相比，TT基因型，CT基因型和CT / TT组合基因型（优势模型）没有改变ESCC的风险。 TT基因型与CT / CC组合基因型的比较也没有显示出与ESCC的显着关联。然而，进一步的分析显示，在主导模型中，食管癌的风险增加（OR = 1.56，95％CI = 1.08-2.26），等位基因频率比较（OR = 1.31，95％CI = 1.06-1.63）在≤60-岁的组。
结论：这些结果表明，miRNA-196a2功能多态性rs11614913可能是中国北方地区ESCC高发地区60岁以下人群进行ESCC风险评估的有效遗传标记。

BACKGROUND & AIMS: BACKGROUND:Increased reactive oxygen species (ROS) production by arsenic treatment in solid tumors showed to be effective to sensitize cancer cells to chemotherapies. Arsenic nano compounds are known to increase the ROS production in solid tumors. METHODS:In this study we developed arsenic-ferrosoferric oxide conjugated Nano Complex (As2S2-Fe3O4, AFCNC) to further promote the ROS induction ability of arsenic reagent in solid tumors. We screen for the molecular pathways that are affect by arsenic treatment in ESCC cancer cells. And explored the underlying molecular mechanism for the arsenic mediated degradations of the key transcription factor we identified in the gene microarray screen. Mouse xenograft model were used to further verify the synthetic effects of AFCNC with chemo and radiation therapies, and the molecular target of arsenic treatment is verified with IHC analysis. RESULTS:With gene expression microarray analysis we found Hippo signaling pathway is specifically affected by arsenic treatment, and induced ubiquitination mediated degradation of YAP in KYSE-450 esophageal squamous cell carcinoma (ESCC) cells. Mechanistically we proved PML physically interacted with YAP, and arsenic induced degradation PML mediated the degradation of YAP in ESCC cells. As a cancer stem cell related transcription factor, YAP 5SA over expressions in cancer cells are correlated with resistance to chemo and radiation therapies. We found AFCNC treatment inhibited the increased invasion and migration ability of YAP 5SA overexpressing KYSE-450 cells. AFCNC treatment also effectively reversed protective effects of YAP 5SA overexpression against cisplatin induced apoptosis in KYSE-450 cells. Lastly, with ESCC mouse xenograft model we found AFCNC combined with cisplatin treatment or radiation therapy significantly reduced the tumor volumes in vivo in the xenograft ESCC tumors. CONCLUSIONS:Together, these findings suggested besides ROS, YAP is a potential target for arsenic based therapy in ESCC, which should play an important role in the synthetic effects of arsenic nano complex with chemo and radiation therapy.

背景与目标： 背景：在实体瘤中通过砷处理增加了活性氧（ROS）的产生，可有效地使癌细胞对化学疗法敏感。已知砷纳米化合物会增加实体瘤中的ROS产生。
方法：在这项研究中，我们开发了砷-铁-铁氧化物共轭纳米复合物（As2S2-Fe3O4，AFCNC），以进一步增强砷剂在实体瘤中的ROS诱导能力。我们筛选了受砷治疗的ESCC癌细胞中影响的分子途径。并探索了我们在基因芯片筛选中鉴定的关键转录因子的砷介导降解的潜在分子机制。使用小鼠异种移植模型进一步验证化学疗法和放射疗法对AFCNC的合成作用，并通过IHC分析验证了砷治疗的分子靶标。
结果：通过基因表达微阵列分析，我们发现了Hippo信号通路受到砷处理的特异性影响，并在KYSE-450食管鳞癌（ESCC）细胞中诱导了泛素化介导的YAP降解。从机理上讲，我们证明了PML与YAP发生了物理相互作用，并且砷诱导的降解PML介导了ESCC细胞中YAP的降解。作为与癌症干细胞相关的转录因子，YAP 5SA在癌细胞中的过度表达与对化学疗法和放射疗法的抵抗力相关。我们发现AFCNC处理抑制了过表达KYSE-450细胞的YAP 5SA的增加的侵袭和迁移能力。 AFCNC处理还有效逆转了YAP 5SA过表达对KYSE-450细胞中顺铂诱导的细胞凋亡的保护作用。最后，在ESCC小鼠异种移植模型中，我们发现AFCNC与顺铂治疗或放射治疗相结合可显着降低异种移植ESCC肿瘤的体内肿瘤体积。
结论：这些发现提示，除ROS外，YAP还可能成为ESCC砷基治疗的潜在靶标，这在化学和放射治疗砷纳米复合物的合成作用中应发挥重要作用。

BACKGROUND & AIMS: :S-adenosylhomocysteine hydrolase (SAHH) is the sole enzyme that catalyses the hydrolysis of S-adenosylhomocysteine (SAH) in methylation reaction. Previous studies have shown that its inhibition or deficiency leads to several human disorders such as severe coagulopathy, hepatopathy and myopathy. However, the effects of SAHH on esophageal squamous cell carcinoma (ESCC) cells have not been explored so far. To determine whether SAHH is involved in carcinogenesis of the esophagus, we investigated the expression of SAHH in ESCC and normal esophageal epithelial cells and found that SAHH was downregulated in ESCC cells compared with normal esophageal epithelial cells (P < 0.05). The overexpressed SAHH in ESCC cells promoted cell apoptosis, inhibited cell migration and adhesion, but did not affect the cell proliferation and cell cycle. Furthermore, an interaction of SAHH with receptor of activated C kinase 1 (RACK1) protein was detected by coimmunoprecipitation and an increased RACK1, which is caused by overexpression of SAHH, was verified by Western blotting. The findings mentioned above demonstrate that SAHH can promote apoptosis, inhibit migration and adhesion of ESCC cells suggesting that it may be involved in carcinogenesis of the esophagus.

背景与目标： ：S-腺苷同型半胱氨酸水解酶（SAHH）是在甲基化反应中催化S-腺苷同型半胱氨酸（SAH）水解的唯一酶。先前的研究表明，其抑制或缺乏会导致多种人类疾病，例如严重的凝血病，肝病和肌病。但是，到目前为止，尚未探讨SAHH对食道鳞状细胞癌（ESCC）细胞的作用。为了确定SAHH是否与食道癌发生有关，我们调查了SAHH在ESCC和正常食管上皮细胞中的表达，发现与正常食管上皮细胞相比SAHH在ESCC细胞中下调（P <0.05）。在ESCC细胞中过表达的SAHH促进细胞凋亡，抑制细胞迁移和粘附，但不影响细胞增殖和细胞周期。此外，通过免疫共沉淀检测到SAHH与活化的C激酶1（RACK1）蛋白受体之间的相互作用，并通过Western印迹法证实了SAHH过表达引起的RACK1增加。上面提到的发现表明SAHH可以促进细胞凋亡，抑制ESCC细胞的迁移和粘附，表明它可能与食道的癌变有关。

BACKGROUND & AIMS: BACKGROUND:The XRCC6 and XRCC5 genes are part of the nonhomologous end-joining (NHEJ) pathway, which is the main mechanism repairing DNA double-strand breaks (DSBs) in human cells. Genetic variations of XRCC6 and XRCC5 might contribute to esophageal squamous cell carcinoma (ESCC) susceptibility. METHODS:ESCC patients (n = 189) and cancer-free controls (n = 189) were recruited in an ESCC high-risk area of north China. Then the rs2267437 (XRCC6), rs3835 (XRCC5) and rs16855458 (XRCC5) polymorphisms were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS:A significant difference in genotype distribution and allele frequency of rs2267437 (XRCC6) was observed between the cases and controls. The CG carriers were at higher risk of ESCC (p = 0.001, odds ratio [OR] = 2.040, 95% confidence interval [95% CI], 1.323-3.147). G allele carriers were also associated with an increased ESCC risk (p = 0.003, OR = 1.868, 95% CI, 1.230-2.836). In the 2 polymorphisms of XRCC5, no significant difference was found between both groups in the distribution of either genotype or allelic frequency. But in the haplotypes established by the single nucleotide polymorphisms (SNPs) of XRCC5, the haplotype AT and CC separately increased by 4.28- and 2.31-fold the risk ratio of ESCC (p = 0.01, OR = 4.28, 95% CI, 1.40-13.05; p = 0.03, OR = 2.31, 95% CI, 1.11-4.80, respectively). In addition, gene-smoking or gene-drinking interactions, and their effect on the risk of ESCC were observed, but no significant gene-environment interaction was demonstrated. CONCLUSIONS:In conclusion, both the CG carriers/G allele carriers of rs2267437 (XRCC6) and the haplotype AT/CC established by the SNPs of XRCC5 are associated with ESCC susceptibility.

背景与目标： 背景：XRCC6和XRCC5基因是非同源末端连接（NHEJ）途径的一部分，该途径是修复人类细胞中DNA双链断裂（DSB）的主要机制。 XRCC6和XRCC5的遗传变异可能有助于食管鳞状细胞癌（ESCC）的易感性。
方法：在中国北方的ESCC高危地区招募ESCC患者（189例）和无癌对照（189例）。然后使用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析对rs2267437（XRCC6），rs3835（XRCC5）和rs16855458（XRCC5）多态性进行基因分型。
结果：病例与对照组之间的rs2267437（XRCC6）基因型分布和等位基因频率存在显着差异。 CG携带者罹患食管鳞癌的风险较高（p = 0.001，优势比[OR] = 2.040，95％置信区间[95％CI]，1.323-3.147）。 G等位基因携带者也与ESCC风险增加相关（p = 0.003，OR = 1.868，95％CI，1.230-2.836）。在XRCC5的2个多态性中，两组在基因型或等位基因频率的分布上均未发现显着差异。但是在由XRCC5的单核苷酸多态性（SNP）建立的单倍型中，单倍型AT和CC分别增加了ESCC风险比的4.28和2.31倍（p = 0.01，OR = 4.28，95％CI，1.40- 13.05； p = 0.03，OR = 2.31，95％CI，1.11-4.80）。此外，观察到基因吸烟或基因饮用相互作用，以及它们对ESCC风险的影响，但未显示出显着的基因-环境相互作用。
结论：总之，rs2267437（XRCC6）的CG携带者/ G等位基因携带者和XRCC5的SNPs建立的单倍型AT / CC均与ESCC易感性相关。

BACKGROUND & AIMS: :The transforming growth factor β (TGF-β) signaling pathway plays anti- and pro-tumoral roles in the vast majority of cancers, and long noncoding RNAs have been reported to play key roles in the highly contextual response process. However, the roles of long noncoding RNAs (lncRNAs) in TGF-β signaling in esophageal squamous cell carcinoma (ESCC) remain unknown. In this study, we performed RNA-seq to compare lncRNAs expression levels between TGF-β1-treated and untreated ESCC cells and observed that NF-kappaB-interacting lncRNA (NKILA) was remarkably upregulated by the classical TGF-β signaling pathway. RNA profiling of 39 pairs ESCC tumor and adjacent nontumor samples using RT-qPCR demonstrated that NKILA is significantly downregulated in ESCC tumor tissues, and NKILA expression levels were significantly decreased in advanced tumor tissues (III and IV) compared to early stages (I and II) (p < 0.01). Gain- and loss-of-function assays showed that NKILA inhibited ESCC cell metastasis in vitro and in vivo, and mechanism studies showed that NKILA repressed MMP14 expression by inhibiting IκBα phosphorylation and NF-κB activation. Collectively, these findings suggest that the TGF-β-induced lncRNA NKILA has potential as an antimetastasis therapy. KEY MESSAGES:Long noncoding RNA NKILA could be remarkably upregulated by classical TGF-β signal pathway in ESCC. NKILA was significantly downregulated in esophageal squamous cell carcinoma and negatively correlated with TNM stage. NKILA inhibits ESCC cell metastasis via repressing MMP14 expression by suppressing the phosphorylation of IκBα and NF-κB activation.

背景与目标： ：转化生长因子β（TGF-β）信号通路在绝大多数癌症中均具有抗肿瘤和促肿瘤作用，据报道，长的非编码RNA在高度相关的反应过程中起关键作用。然而，在食管鳞状细胞癌（ESCC）中，长非编码RNA（lncRNA）在TGF-β信号传导中的作用仍然未知。在这项研究中，我们进行了RNA序列比较以比较TGF-β1处理和未处理的ESCC细胞之间的lncRNA表达水平，并观察到NF-κB相互作用的lncRNA（NKILA）被经典的TGF-β信号通路显着上调。使用RT-qPCR对39对ESCC肿瘤和邻近非肿瘤样品进行RNA谱分析表明，与早期阶段（I和II）相比，NKILA在ESCC肿瘤组织中显着下调，并且在晚期肿瘤组织（III和IV）中NKILA表达水平显着降低）（p <0.01）。功能获得和丧失功能测定显示NKILA在体内外均抑制ESCC细胞转移，机理研究表明NKILA通过抑制IκBα磷酸化和NF-κB活化来抑制MMP14表达。总的来说，这些发现表明TGF-β诱导的lncRNA NKILA具有作为抗转移疗法的潜力。
关键信息：ESCC中的经典TGF-β信号通路可能显着上调了长非编码RNA NKILA。 NKILA在食管鳞状细胞癌中显着下调，与TNM分期呈负相关。 NKILA通过抑制IκBα的磷酸化和NF-κB的活化来抑制MMP14的表达，从而抑制ESCC细胞的转移。

BACKGROUND & AIMS: :NF45 (also known as ILF2), as one subunit of NF-AT (nuclear factor of activated T cells), repairs DNA breaks, inhibits viral replication, and also functions as a negative regulator in the microRNA processing pathway in combination with NF90. Recently, it was found that implicated in the mitotic control of HeLa cells and deletion of endogenous NF45 decreases growth of HeLa cells. While the role of NF45 in cancer biology remains under debate. In this study, we analyzed the expression and clinical significance of NF45 in esophageal squamous cell carcinoma ESCC. The expression of NF45 was evaluated by Western blot in 8 paired fresh ESCC tissues and immunohistochemistry on 105 paraffin-embedded slices. NF45 was highly expressed in ESCC and significantly associated with ESCC cells tumor stage and Ki-67. Besides, high NF45 expression was an independent prognostic factor for ESCC patients' poor survival. To determine whether NF45 could regulate the proliferation of ESCC cells, we increased endogenous NF45 and analyzed the proliferation of TE1 ESCC cells using Western blot, CCK8, flow cytometry assays and colony formation analyses, which together indicated that overexpression of NF45 favors cell cycle progress of TE1 ESCC cells. While knockdown of NF45 resulted in cell cycle arrest at G0/G1-phase and thus abolished the cell growth. These findings suggested that NF45 might play an important role in promoting the tumorigenesis of ESCC, and thus be a promising therapeutic target to prevent ESCC progression.

背景与目标： ：NF45（也称为ILF2），作为NF-AT（活化的T细胞的核因子）的一个亚基，修复DNA断裂，抑制病毒复制，并与NF90一起在microRNA加工途径中起负调控作用。最近，发现牵涉HeLa细胞的有丝分裂控制和内源性NF45的缺失降低HeLa细胞的生长。尽管NF45在癌症生物学中的作用仍在争论中。在这项研究中，我们分析了NF45在食管鳞状细胞癌ESCC中的表达及其临床意义。通过Western印迹法在8对配对的ESCC组织中对NF45的表达进行了评估，并在105个石蜡包埋的切片上进行了免疫组织化学分析。 NF45在ESCC中高表达，并与ESCC细胞的肿瘤分期和Ki-67显着相关。此外，NF45高表达是ESCC患者生存不良的独立预后因素。为了确定NF45是否能调节ESCC细胞的增殖，我们增加了内源性NF45并使用Western blot，CCK8，流式细胞术和集落形成分析来分析TE1 ESCC细胞的增殖，这共同表明NF45的过表达有利于NF45的细胞周期进程。 TE1 ESCC细胞。而敲低NF45导致细胞周期停滞在G0 / G1期，从而废除了细胞生长。这些发现表明NF45可能在促进ESCC的肿瘤发生中起重要作用，因此是预防ESCC进展的有希望的治疗靶标。

BACKGROUND & AIMS: OBJECTIVE:To determine whether the combination of prostaglandin E2 (PGE2) and EP2, the subtype receptor of PGE2, could trans-activate the epidermal growth factor receptor (EGFR). PATIENTS AND METHODS:In this experiment, we selected epithelial cells from normal esophageal mucosa as the negative control group, and the ESCC EC109 and TE-1 cell strain as the observation group. Real-time PCR and Western-blotting were used to detect the expression of EP2, EGFR and phosphorylated EGFR (p-EGFR). The pre-treatment of ESCC cell strains was carried out using Butaprost (special agonist of PGE2 and EP2) and RNAi of EP2, and we observed the expression of EP2, EGFR, and p-EGFR. WST-8 (CCK-8) was applied for the detection of the cell proliferation rate. The transwell invasion experiment was conducted for the detection of the invasion capability of cells. The expression of MMP-9 (matrix metalloproteinase-9), VEGF (vascular endothelial growth factor), pro-inflammatory factors (IL-6 and TNF-α) in the cell supernatant were detected using ELISA. RESULTS:The high mRNA and protein expression of EP2, EGFR, and p-EGFR were found in the EC109 and TE-1 cell strains in the observation group, which were higher than those in the control group (p < 0.05). After the intervention of PGE2, EP2 expression was decreased and the p-EGFR expression was increased (p < 0.05). There was no variation found in the expression of EGFR (p > 0.05). After cells were intervened using Butaprost, the expressions of EP2 and p-EGFR were increased (p < 0.05), and there were no changes identified in the expression of EGFR (p > 0.05). After the intervention of RNAi, the expression of EP2 and p-EGFR was decreased (p < .05), and no changes were identified in the expression of EGFR (p > 0.05). After the intervention of PGE2 and Butaprost, great increases were seen in the cell proliferation rate, invasion capability, and the expression of MMP-9, VEGF, IL-6, and TNF-α in EC109 and TE-1 cell strains (p < 0.05), however, the intervention of RNAi could reduce above indexes (p < 0.05). CONCLUSIONS:Through cell experiments, we verified that the combination of prostaglandin E2 (PGE2) and EP2, the subtype receptor of PGE2, could trans-activate the epidermal growth factor receptor (EGFR) to regulate the proliferation and invasion capability of esophageal squamous cell carcinoma (ESCC) cells, and secrete and express multiple cytokines, thus discovering the pathological mechanism of inflammation to carcinoma transition in the occurrence of ESCC, and providing the experimental evidence for the search of new target in the treatment of ESCC. ESCC cells can highly express the receptor subtype EP2 of PGE2 that can transactivate the EGFR, through which PGE2 is involved in the transition mechanism from inflammation to cancer.

背景与目标： 目的：确定前列腺素E2（PGE2）和EP2（PGE2的亚型受体）的组合是否可以反激活表皮生长因子受体（EGFR）。
病人与方法：本实验以正常食管粘膜上皮细胞为阴性对照组，以ESCC EC109和TE-1细胞株为观察组。实时PCR和Western-blotting用于检测EP2，EGFR和磷酸化EGFR（p-EGFR）的表达。使用Butaprost（PGE2和EP2的特殊激动剂）和EP2的RNAi进行ESCC细胞株的预处理，我们观察到EP2，EGFR和p-EGFR的表达。 WST-8（CCK-8）用于检测细胞增殖率。进行transwell侵袭实验以检测细胞的侵袭能力。使用ELISA检测细胞上清液中MMP-9（基质金属蛋白酶-9），VEGF（血管内皮生长因子），促炎因子（IL-6和TNF-α）的表达。
结果：观察组EC109和TE-1细胞株中EP2，EGFR和p-EGFR的mRNA和蛋白表达较高，高于对照组（p <0.05）。在PGE2干预后，EP2表达降低，而p-EGFR表达升高（p <0.05）。在EGFR的表达中没有发现变化（p＞ 0.05）。用Butaprost干预细胞后，EP2和p-EGFR的表达增加（p <0.05），而EGFR的表达没有变化（p> 0.05）。 RNAi干预后，EP2和p-EGFR的表达降低（p <.05），而EGFR的表达未见变化（p> 0.05）。在PGE2和Butaprost的干预下，EC109和TE-1细胞株的细胞增殖速率，侵袭能力以及MMP-9，VEGF，IL-6和TNF-α的表达均显着增加（p < 0.05），但是，RNAi的干预可以降低上述指标（p <0.05）。
结论：通过细胞实验，我们证实前列腺素E2（PGE2）和EP2（PGE2的亚型受体）的组合可以反激活表皮生长因子受体（EGFR）来调节食管鳞状细胞癌的增殖和侵袭能力。 （ESCC）细胞，并分泌和表达多种细胞因子，从而发现了发生ESCC时炎症向癌变的病理机制，为寻找ESCC的新靶标提供了实验依据。 ESCC细胞可以高表达PGE2的受体亚型EP2，该亚型可以使EGFR活化，PGE2可以通过该亚型参与从炎症到癌症的转变机制。

BACKGROUND & AIMS: Background:Radiotherapy is one of the most important treatments for esophageal squamous cell carcinoma (ESCC). Previously, we found that EphA5 expression was increased in ESCC cells and tumor tissues. Studies from other groups reported that EphA5 is abnormally expressed in numerous malignant tumors and may be involved in the radiosensitivity of lung cancer. However, the role of EphA5 in radiotherapy for ESCC remains unclear. Methods:The siRNA sequences against human EPHA5 were transfected to the ESCC cells (KYSE150 and KYSE450). After ionizing radiation (IR), cell viability and colony formation assays were used to test the changes of cell proliferation in EphA5-silenced cells. Flow cytometry analysis was performed to investigate the cell apoptosis and cycle in the irradiated cells interfered by siRNA. The key molecules involved in cell cycle checkpoints and DNA damage repair were evaluated by Western blot and immunofluorescence. Results:CCK8 assay and clonogenic assay showed that the proliferation of EphA5-silenced ESCC cells was inhibited after IR. At 24 h post-IR treatment, we found that the G1/S checkpoint triggered by DNA damage in EphA5-silenced cells was defective. γ-H2AX foci in the irradiated EphA5-silenced cells were impaired at 0.5 h post-IR treatment as well as ATM activation. The defective activation of ATM resulted in a decrease of p-Chk2, p-p53 and p21 expression. Conclusion:In conclusion, these results indicate that EphA5 silencing increases radiosensitivity in ESCC cells through ATM-dependent pathway, which provides a potential target for the radiotherapy in ESCC.

背景与目标： 背景：放射治疗是食管鳞状细胞癌（ESCC）的最重要治疗方法之一。以前，我们发现EphA5表达在ESCC细胞和肿瘤组织中增加。其他研究表明，EphA5在许多恶性肿瘤中异常表达，可能与肺癌的放射敏感性有关。然而，EphA5在ESCC放射治疗中的作用尚不清楚。
方法：将针对人类EPHA5的siRNA序列转染至ESCC细胞（KYSE150和KYSE450）。电离辐射（IR）后，细胞活力和集落形成测定法用于测试EphA5沉默的细胞中细胞增殖的变化。进行流式细胞术分析以研究被siRNA干扰的被照射细胞的细胞凋亡和周期。通过蛋白质印迹和免疫荧光评估了参与细胞周期检查点和DNA损伤修复的关键分子。
结果：CCK8法和克隆形成法显示，IR后，EphA5沉默的ESCC细胞的增殖受到抑制。红外线治疗后24小时，我们发现由EphA5沉默的细胞中的DNA损伤触发的G1 / S检查点存在缺陷。照射后的EphA5沉默的细胞中的γ-H2AX焦点在IR处理以及ATM激活后0.5小时受到损害。 ATM激活缺陷导致p-Chk2，p-p53和p21表达降低。
结论：总之，这些结果表明，EphA5沉默可通过ATM依赖性途径提高ESCC细胞的放射敏感性，这为ESCC放射治疗提供了潜在的靶标。

BACKGROUND & AIMS: :Tumor associated macrophages (TAMs) play an important role in the growth, progression, and metastasis of tumors. The distribution of TAMs in Kazakh esophageal squamous cell carcinoma (ESCC) is not determined. We aimed to investigate the role of TAMs in the occurrence and progression of Kazakh ESCC. CD163 was used as the TAM marker, and immunohistochemistry (IHC) counts were used to quantify the density of TAMs in tumor nest and surrounding stroma. IHC staining was used to evaluate the expression of vascular endothelial growth factor C (VEGF-C) in Kazakh ESCC and cancer adjacent normal (CAN) tissues. The density of TAMs in Kazakh ESCCs tumor nest and stromal was significantly higher than that in CAN tissues. The increased number of CD163-positive TAMs in tumor nest and tumor stromal was positively associated with Kazakh ESCC lymph node metastasis and clinical stage progression. Meanwhile, the expression of VEGF-C in Kazakh ESCCs was significantly higher than that in CAN tissues. Overexpression of VEGF-C in Kazakh ESCCs was significantly associated with gender, depth of tumor invasion, lymph node metastasis and tumor clinical stage. The increased number of TAMs, either in the tumor nests or tumor stroma was positively correlated with the overexpression of VEGF-C, which may promote lymphangiogenesis and play an important role in the invasion and metastasis of Kazakh ESCC.

背景与目标： ：肿瘤相关的巨噬细胞（TAM）在肿瘤的生长，进展和转移中起重要作用。在哈萨克人食管鳞状细胞癌（ESCC）中TAM的分布尚未确定。我们旨在调查TAM在哈萨克ESCC发生和发展中的作用。 CD163用作TAM标记，免疫组化（IHC）计数用于量化肿瘤巢和周围基质中TAM的密度。 IHC染色用于评估哈萨克斯坦ESCC和癌旁正常组织（CAN）中血管内皮生长因子C（VEGF-C）的表达。哈萨克ESCCs肿瘤巢和基质中TAMs的密度显着高于CAN组织中的TAMs的密度。肿瘤巢和肿瘤基质中CD163阳性TAM数量的增加与哈萨克斯坦ESCC淋巴结转移和临床分期进展呈正相关。同时，哈萨克人食管鳞癌中VEGF-C的表达明显高于CAN组织。哈萨克人食管鳞癌中VEGF-C的过度表达与性别，肿瘤浸润深度，淋巴结转移和肿瘤临床分期显着相关。无论是在肿瘤巢中还是在肿瘤基质中，TAM的数量增加都与VEGF-C的过表达呈正相关，这可能促进淋巴管生成，并在哈萨克ESCC的侵袭和转移中起重要作用。

BACKGROUND & AIMS: :Esophageal squamous cell carcinoma (ESCC) is one of the most malignant gastrointestinal cancers and occurs at a high frequency rate in China and other Asian countries. Recently, several molecular markers were identified for predicting ESCC. Notwithstanding, additional prognostic markers, with a clear understanding of their underlying roles, are still required. Through bioinformatics, a graph-clustering method by DPClus was used to detect co-expressed modules. The aim was to identify a set of discriminating genes that could be used for predicting ESCC through graph-clustering and GO-term analysis. The results showed that CXCL12, CYP2C9, TGM3, MAL, S100A9, EMP-1 and SPRR3 were highly associated with ESCC development. In our study, all their predicted roles were in line with previous reports, whereby the assumption that a combination of meta-analysis, graph-clustering and GO-term analysis is effective for both identifying differentially expressed genes, and reflecting on their functions in ESCC.

背景与目标： 食管鳞状细胞癌（ESCC）是最恶性的胃肠道癌之一，在中国和其他亚洲国家以很高的频率发生。最近，鉴定了几种分子标志物来预测ESCC。尽管如此，仍然需要其他的预后指标，并清楚地了解其潜在作用。通过生物信息学，使用DPClus的图聚类方法检测共表达的模块。目的是确定一组可用于通过图聚类和GO项分析预测ESCC的区分基因。结果表明，CXCL12，CYP2C9，TGM3，MAL，S100A9，EMP-1和SPRR3与ESCC的发展高度相关。在我们的研究中，它们的所有预测作用均与以前的报告一致，因此，假设荟萃分析，图聚类和GO项分析相结合可有效识别差异表达的基因并反映其在ESCC中的功能。

BACKGROUND & AIMS: :CtBP2, as a transcriptional corepressor of epithelial-specific genes, has been reported to promote tumor due to upregulating epithelial-mesenchymal transition (EMT) in cancer cells. CtBP2 was also demonstrated to contribute to the proliferation of esophageal squamous cell carcinoma (ESCC) cells through a negative transcriptional regulation of p16(INK4A). In this study, for the first time, we reported that CtBP2 expression, along with CCNH/CDK7, was higher in ESCC tissues with lymph node metastases than in those without lymph node metastases. Moreover, both CtBP2 and CCNH/CDK7 were positively correlated with E-cadherin, tumor grade, and tumor metastasis. However, the concrete mechanism of CtBP2's role in enhancing ESCC migration remains incompletely understood. We confirmed that CCNH/CDK7 could directly interact with CtBP2 in ESCC cells in vivo and in vitro. Furthermore, our data demonstrate for the first time that CtBP2 enhanced the migration of ESCC cells in a CCNH/CDK7-dependent manner. Our results indicated that CCNH/CDK7-CtBP2 axis may augment ESCC cell migration, and targeting the interaction of both may provide a novel therapeutic target of ESCC.

背景与目标： ：CtBP2，作为上皮特异性基因的转录共抑制子，由于在癌细胞中上皮-间质转化（EMT）上调而促进了肿瘤。还证明了CtBP2通过对p16（INK4A）的负转录调节来促进食管鳞状细胞癌（ESCC）细胞的增殖。在本研究中，我们首次报道，在有淋巴结转移的ESCC组织中，CtBP2表达以及CCNH / CDK7高于无淋巴结转移的ESCC组织。此外，CtBP2和CCNH / CDK7与E-钙粘蛋白，肿瘤等级和肿瘤转移均呈正相关。但是，对CtBP2在增强ESCC迁移中作用的具体机制尚不完全了解。我们证实，CCNH / CDK7可以在体内和体外直接与ESCC细胞中的CtBP2相互作用。此外，我们的数据首次证明CtBP2以CCNH / CDK7依赖性方式增强了ESCC细胞的迁移。我们的研究结果表明CCNH / CDK7-CtBP2轴可能会增加ESCC细胞的迁移，并且靶向两者的相互作用可能会提供ESCC的新型治疗靶点。