TSLC1在食管鳞癌中的表达及临床意义

目的：研究食管鳞状细胞癌组织中肺癌肿瘤抑制因子1（TSLC1）基因的表达及临床病理意义,探讨其在食管鳞状细胞癌发生、发展过程中的作用。方法：应用免疫组化（SP法）检测50例食管鳞状细胞癌标本及癌旁正常食管组织中TSLC1基因的表达情况,分析其与食管鳞状细胞癌患者临床病理学参数之间的关系。结果：50例食管鳞状细胞癌组织中TSLC1基因表达阳性率为48.0%（24/50）,其表达与癌组织浸润深度、区域淋巴结转移及TNM分期有关（P〈0.05）。与性别、年龄、肿瘤大体分型、肿瘤大小、位置、分化程度无关（P〉0.05）。50例癌旁正常食管组织中TSLC1基因高表达,阳性率为94.0%（47/50）。食管鳞状细胞癌组织中的阳性表达率显著低于癌旁正常食管组织,两者相比差异有统计学意义（P〈0.01）。结论：食管鳞状细胞癌组织中有明显的TSLC1基因的表达缺失,TSLC1蛋白表达缺失与食管鳞状细胞癌的发生、发展有关。     

Promoter hypermethylation of CDH13 is a common, early event in human esophageal adenocarcinogenesis and correlates with clinical risk factors

Although the CDH13 gene has been shown to undergo epigenetic silencing by promoter methylation in many types of tumors, hypermethylation of this gene in Barrett's-associated esophageal adenocarcinogenesis has not been studied. Two hundred fifty-nine human esophageal tissues were therefore examined for CDH13 promoter hypermethylation by real-time methylation-specific PCR. CDH13 hypermethylation showed discriminative receiver-operator characteristic curve profiles, sharply demarcating esophageal adenocarcinoma (EAC) from esophageal squamous cell carcinoma (ESCC) and normal esophagus (NE) (p < 0.0001). CDH13 normalized methylation values (NMV) were significantly higher in Barrett's esophagus (BE), dysplastic BE (D) and EAC than in NE (p < 0.0000001). CDH13 hypermethylation frequency was 0% in NE but increased early during neoplastic progression, rising to 70% in BE, 77.5% in D and 76.1% in EAC. Both CDH13 hypermethylation frequency and its mean NMV were significantly higher in BE with than without accompanying EAC. In contrast, only 5 (19.2%) of 26 ESCCs exhibited CDH13 hypermethylation. Furthermore, both CDH13 hypermethylation frequency and its mean NMV were significantly higher in EAC than in ESCC, as well as in BE or D vs. ESCC. Interestingly, mean CDH13 NMV was significantly lower in short-segment than in long-segment BE, a known clinical risk factor for neoplastic progression. Similarly, BE segment length was significantly lower in specimens with unmethylated than with methylated CDH13 promoters. 5-aza-2'-deoxycytidine treatment of OE33 EAC and KYSE220 ESCC cells reduced CDH13 methylation and increased CDH13 mRNA expression. These findings suggest that hypermethylation of CDH13 is a common, tissue-specific event in human EAC, occurs early during BE-associated neoplastic progression, and correlates with known clinical neoplastic progression risk factors.     

食管鳞癌中血管内皮生长因子－C 的表达及与淋巴管生成的关系

目的：探讨血管内皮生长因子－C（VEGF －C）在食管鳞癌中的表达及其与淋巴管生成、淋巴结转移的关系。方法：收集2013年3月至2014年1月遂宁市中心医院胸心外科的107例食管鳞癌手术切除病例及56例正常食管组织的石蜡包埋组织样本,采用免疫组化检测 VEGF －C 在食管鳞癌中的表达,并应用 D2－40抗体标记组织中的微淋巴管内皮细胞,计数微淋巴管密度（LVD）。同时对其与临床病理参数的关系进行分析。结果：食管鳞癌中 VEGF －C 蛋白的高表达率明显高于正常食管组织,且在食管鳞癌不同 TNM分期中有差异（P ＜0．05）;淋巴结转移组中 VEGF －C 蛋白的高表达率为68．3％,高于无淋巴结转移组（P ＜0．05）。T3／T4期组中 VEGF －C 蛋白的高表达率高于 T1／T2期组（P ＜0．05）。VEGF －C 蛋白的高表达率与食管鳞癌患者的年龄、性别、分化程度、肿瘤大小等均无明显相关性（P ＞0．05）。食管鳞癌中 LVD 在 VEGF －C 不同表达强度组中有差异,差异有统计学意义（P ＜0．05）。食管鳞癌淋巴结转移组 LVD 高于无淋巴结转移组,差异均有统计学意义（P ＜0．05）。结论：VEGF －C 基因可能促进食管鳞癌的淋巴管生成及淋巴结转移,有可能成为预测食管鳞癌预后的实验室指标。     

脾酪氨酸激酶与食管鳞状细胞癌的关系

目的：探讨脾酪氨酸激酶（spleen tyrosine kinase,Syk）在食管鳞状细胞癌组织中的表达及其临床意义。方法：采用逆转录-聚合酶链反应（RT-PCR）检测30例食管鳞癌组织、27例食管鳞癌癌旁组织及30例相对正常食管黏膜组织中Syk mRNA的表达;应用免疫组化技术检测39例食管鳞癌组织、27例食管鳞癌癌旁组织及38例相对正常食管黏膜组织中Syk蛋白的表达并分析其与临床病理学指标间的关系。结果：食管鳞癌组织中Syk mRNA表达高于癌旁组织和正常食管黏膜组织,但差异无统计学意义（P〉0.05）。Syk蛋白在食管鳞癌组织中的阳性率（34/39,87.2%）明显高于食管鳞癌癌旁组织（18/27,66.7%）和正常食管黏膜组织（26/38,68.4%）（P〈0.05）。Syk蛋白表达与食管鳞癌患者年龄、性别、肿瘤大小、临床分期均无明显相关关系（P均〉0.05）。结论：Syk mRNA和蛋白在食管鳞癌组织中表达增高,但Syk蛋白表达与临床病理学指标无相关关系。     

食管癌耐阿霉素EC9706中干细胞富集及上皮间质化

目的:建立耐阿霉素(ADM)食管癌细胞株,命名为EC9706/ADM,探讨该细胞株的上皮间质化(EMT)及肿瘤干细胞富集现象.方法:用中等浓度间歇法建立食管癌耐阿霉素细胞株,MTT法测定细胞株对ADM、5-FU、顺铂、环磷酰胺的耐药指数.流式细胞仪检测EC9706/ADM及EC9706中CD44+CD24-细胞比例,MTT法、Transwell、平板克隆形成和球囊培养实验分别检测耐药对细胞增殖、迁移与转移、体外成瘤和克隆形成能力的影响;荧光定量PCR检测EC9706/ADM干细胞标志ALDH1、CD133及Oct4及EMT相关分子标志物E-钙黏蛋白及间质细胞分子标记N-钙黏蛋白、波形蛋白、纤维连接蛋白的表达.结果:EC9706/ADM的耐药指数为17.8,对5-FU、顺铂、环磷酰胺产生了交叉耐药性.与EC9706细胞相比,EC9706/ADM耐药细胞转移能力明显增强,表现出更高的微球体形成能力,E-cadherin表达降低而Fibronectin表达显著增高.结论:食管癌耐阿霉素细胞EC9706/ADM发生了EMT现象,转移迁移能力增强,细胞表现出更高的干细胞特性,化疗药物有助于肿瘤干细胞的富集.     

LETM2在食管鳞癌中的表达及其作用

  目的  探讨LETM2在食管鳞癌（esophageal squamous cell carcinoma,ESCC）中的表达,研究LETM2对ESCC细胞系KYSE150和ECA109增殖和侵袭迁移的影响。  方法  免疫组化（immunohistochemistry,IHC）检测90例ESCC及癌旁组织LETM2蛋白表达差异,RT-PCR和Western blot检测ESCC细胞系LETM2表达情况,慢病毒敲低KYSE150和ECA109细胞中LETM2表达。MTT和克隆形成技术检测LETM2对ESCC细胞增殖和克隆形成能力的影响,流式细胞术检测细胞周期变化,Transwell法分析LETM2敲低后细胞迁移侵袭能力的差异。  结果  在ESCC组织中LETM2的表达显著高于癌旁组织,LETM2通过抑制ESCC细胞G₁期向S期转化进而抑制细胞增殖,但对侵袭和迁移能力影响不大。  结论  LETM2可能作为驱动基因促进ESCC的发生发展,是ESCC的关键遗传变异,可能作为ESCC早诊早治的分子标志物。      

食管鳞状细胞癌中miRNAs的研究进展

食管鳞状细胞癌（esophageal squamous cell carcinoma, ESCC）是我国常见食管癌（esophageal cancer, EC）的主要亚型。虽然ESCC从分子遗传学角度已被广泛研究,但其发病分子机制还没有完全阐明。microRNAs（miRNAs）是一类小的内源性非蛋白编码RNAs,在生理、病理过程中发挥着重要作用,包括细胞分化、凋亡、增殖和代谢。miRNAs通过致癌基因、抑癌基因在ESCC的发病机制中扮演着重要角色。近年来研究表明,许多miRNAs在ESCC组织中的表达存在明显的差异,了解miRNAs及其靶基因在ESCC发生发展中的作用,可能为早期检测、诊断、治疗和预后提供策略。     

RFT2 is overexpressed in esophageal squamous cell carcinoma and promotes tumorigenesis by sustaining cell proliferation and protecting against cell death

Human riboflavin transporter 2 (RFT2, also termed as SLC52A3) was recently identified as a susceptibility gene to esophageal squamous cell carcinoma (ESCC), however, its expression and biologic function has remained unclear in ESCC. In this study, we demonstrated that RFT2 was frequently overexpressed in tumor samples compared with normal adjacent tissue in ESCC patients. Knockdown of RFT2 in ESCC cells resulted in decreases of intracellular flavin status, mitochondrial membrane potential and cellular ATP levels, and inhibitions of cell proliferation, colony formation and anchorage-independent growth. Knockdown of RFT2 increased p21 and p27 protein levels, decreased their downstream targets cyclin E1 and Cdk2 protein levels and caused pRb hypophosphorylation, leading to cell cycle arrest at G1-G1/S. Knockdown of RFT2 also reduced anti-apoptotic proteins Bcl-2, Bcl-xl and survivin levels, caused activation of caspase-3 and apoptosis. In contrast, ectopic overexpression of RFT2 in ESCC cells promoted cell proliferation under restricted conditions (soft agar), conferred resistance to cisplatin, and enhanced tumorigenicity in nude mice. These results suggest that RFT2 contributes to ESCC tumorigenesis and may serve as a potential therapeutic target.     

食管鳞状细胞癌组织中KiSS-1的表达及其临床意义

目的 探讨食管鳞状细胞癌（ESCC）组织中肿瘤转移抑制基因（KiSS-1）的表达并分析其临床意义。方法 采用实时定量PCR方法检测65例ESCC组织和同一患者的邻近正常食管组织中KiSS-1的表达及分析其与各种临床病理因数的关系。结果 KiSS-1在肿瘤组织中的表达明显低于正常食管组织(P=0.017)。同时,KiSS-1的表达缺失与肿瘤的大小、分化程度与浸润程度之间无明显的相关性(P=0.480,P=0.827,P=0.823),而与肿瘤淋巴结的转移相关。另外, KiSS-1表达缺失的患者五年生存率相对更差(P=0.009)。结论 KiSS-1表达缺失可作为ESCC发生淋巴结转移的一个指标,KiSS-1也是判断ESCC患者预后的一个指标。