人食管鳞状细胞癌裸小鼠移植瘤模型的研究

将1例人食管癌手术切除组织移植于NIH裸小鼠皮下，建立了人食管鳞状细胞癌裸小鼠移植瘤模型（Eca910709），已传25代，历时2年余传代移植成功率100％。该移植瘤病理组织学的结构与原人体肿瘤的形态相似．电镜形态属鳞状细胞癌。用地高辛标记的Alu探针及DNA－DNA原位杂交方法，证实移植癌中有Alu序列存在，表明移植瘤来源于人类组织。该移植瘤的建立将有助于食管癌病因、发病机理及治疗的研究．     

人卵巢小细胞癌裸小鼠移植瘤细胞超微结构研究

对我所建立的人体卵巢小细胞癌裸小鼠移植瘤模型的各代移植瘤组织进行超微结构观察。发现除光镜原已看到的大、小细胞外,尚有第三种细胞。后者有核畸变,呈多形性等特征,一般大小介于大、小细胞之间,故名中间细胞。分析了三种细胞在超微结构上之异同,并讨论它们之间可能存在的关系。     

平阳霉素对人食管癌裸鼠移植瘤抑瘤作用和耐药性的研究

作平阳霉素(BLMA_5)对人食管鳞状细胞癌裸小鼠移植瘤(Eca910709)进行治疗,按15mg/kg体重腹腔注射给药,经重复实验,结果抑瘤率为58％～70％(P<0.01),说明BLMA_5对Eca910709有明显的抑瘤作用。经三个疗程治疗,Eca910709即对BLMA_5产生耐药,表现在抑瘤率从70％(P<0.01)递减为34％(P<0.05)和6％(P>0.05)。给药剂量增加至30mg/kg,BLMA_5对己耐药的Eca910709再次显示一定的抑瘤作用(抑瘤率为44％,P<0.01)。本实验结果对临束治疗食管癌将有较大应用价值。     

食管鳞状细胞癌研究模型：细胞系、动物移植模型和3D培养模型

食管癌是常见的消化道恶性肿瘤,易转移及复发的性质常常导致病人呈现出明显的致死性风险。国内一些地区食管鳞状细胞癌（esophageal squamous cell carcinoma,ESCC）的发病率相对较高,严重威胁着人类的健康和生命而成为肿瘤研究的重要方向。食管鳞状细胞癌的发生发展是一系列因素累积及相互作用的复杂过程,其克隆性增殖会造成肿瘤细胞内微观水平的表达异常和信号传导通路级联反应效应的改变,形成具有不同性质的细胞亚群。肿瘤细胞间不同细胞亚群在细胞分子水平甚至基因表达层面呈现出比较明显的异质性,由此引发肿瘤细胞对药物的敏感性下降及抗药性现象的出现。很多食管癌移植模型被用于了解肿瘤的性质和病理生理学特点,有助于探索肿瘤细胞内的发生机制,为推动肿瘤研究和治疗提供了重要的价值。随着肿瘤类器官培养技术获得突破性进展,肿瘤细胞的3D培养模型能够重现亲代肿瘤细胞的生物学性质,成为探索肿瘤发生机制和基因序列异常表达的工具,为肿瘤的研究提供了重要的平台,并推动肿瘤研究和治疗向前迈进及取得重要进展。本文对常用的ESCC的研究模型作一综述。     

食管鳞状细胞癌患者γδT细胞研究

 目的 探讨食管癌患者机体γδT细胞水平及与临床的关系，为γδT细胞在食管癌综合治疗中的应用提供依据。方法 采用流式细胞术测定食管癌患者外周血中和癌组织中γδT细胞水平，分析其与临床的关系。结果 癌组织中浸润γδT细胞水平显著高于癌旁组织（P = 0.045）；食管癌患者外周血中γδT细胞水平显著低于正常人（P = 0.011）；食管癌术后患者外周血中γδT细胞水平显著高于术前患者（P = 0.037）。食管癌T分期、N分期、M分期、大体分型、病变部位、病变长度（3 ~ 5 cm组和5 cm组）、组织学分级各组与正常组比较差异有统计学意义（P＜0.05）。食管癌患者外周血中总T细胞，各组与正常组比较差异无统计学意义（P＞0.05）；外周血中γδT细胞年龄、性别、病变长度（＜3 cm），各组与正常组比较差异无统计学意义（P＞0.05）。结论 γδT细胞在机体抗食管癌中发挥重要作用，手术切除肿瘤后可使患者γδT细胞恢复正常水平。     

Shh和Ptch在食管鳞状细胞癌中的表达研究

目的探讨食管鳞状细胞癌中Shh和Ptch的表达及其意义。方法应用免疫组织化学方法检测35例食管癌及癌旁组织中shh和Ptch的表达情况。结果食管鳞癌中Shh和Ptch的表达阳性率分别为74．3％和68．6％，与正常组织相比Shh和Ptch在食管鳞癌组织中高表达。结论Shh和Ptch的高表达及Hh通路的激活参与了食管鳞癌的发生，可能足食管癌治疗的一个理想靶标。     

食管鳞状细胞癌中ERK的表达及其临床意义

目的:研究食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)及癌旁黏膜组织中的细胞外信号调节激酶(extracellular signal ragulated kinase,ERK)mRNA的表达,探讨ERK在ESCC发生、发展中的作用及其意义。方法:收集河北医科大学第四医院胸外科40例ESCC患者癌组织标本及25例患者的癌旁组织标本,RT-PCR检测ESCC与癌旁组织中ERK mRNA的表达情况,分析其与ESCC临床病理特征的关系。结果:ESCC组织中ERK mRNA的表达明显增高,而在癌旁组织中呈现低表达(0.656±0.055 vs 0.450±0.070,P<0.01)。ESCC组织中ERK mRNA的表达与ESCC患者的年龄、性别和组织分化程度无关(P>0.05),而与临床分期、淋巴结转移显著相关(P<0.05)。结论:ESCC组织高表达ERK mRNA,ERK可能在ESCC发生、发展中起重要的作用。     

人结肠高分化腺癌裸小鼠移植瘤模型的建立及其生物学特性

将人结肠高分化腺癌手术标本，接种于裸小鼠体内建成移植瘤ＴＨＣＮ－８９０１，已传１４代，历时２年多，生长稳定，可移植性成功率高。组织学及超微结构形态观察，均保持了原病人肿瘤的结构，并且有分泌功能，染色体分析证实为人染色体，众数在６３～９４之间，该瘤株具有稳定产生ＣＥＡ的生物学特性，用＾１１１Ｉｎ－ＣＥＡＭｃＡｂ能使裸小鼠移植瘤清晰显像。此瘤株的建立为结肠癌癌变基础及实验性基因治疗研究，提供了较为理想     

食管鳞状细胞癌的肿瘤血管生成机制

血管内皮生长因子、缺氧诱导因子、白细胞介素、血管生成素样蛋白、整合素和上皮间质转化在食管鳞状细胞癌血管生成中为癌细胞的生长、侵袭和转移提供了营养支持和有利环境.对食管鳞状细胞癌血管生成机制的研究将为其抗血管靶向治疗提供更多思路和潜在靶点.     

p16蛋白在食管鳞癌中的表达及其意义

目的：研究p16蛋白表达与食管鳞癌生物学行为的关系。方法：应用免疫组化LSAB法检测52例食管鳞癌p16蛋白表达。结果：21例食管鳞癌p16蛋白表达阳性。阳性率为40.4%，随着恶性程度增加及病程进展，p16蛋白阳性率逐渐下降；p16蛋白阳性与淋巴结转移有关。p16蛋白阴性患者死亡率明显高于p16蛋白阳性者，且存活时间短。结论：检测食管鳞癌组织p16蛋白表达，有助于综合判断食管鳞癌恶性程度，转移潜能和预后。     

耐紫杉醇食管鳞癌细胞株Ec9706/P-1的建立及特征

目的：建立人食管癌紫杉醇耐药细胞系Ec9706/P-1并研究其生物学特征,为食管癌临床治疗提供实验依据.方法：小剂量逐步间歇诱导,建立人食管癌紫杉醇耐药细胞系Ec9706/P-1.用MTT法检测该耐药细胞株的多药耐药性及耐药指数,采用流式细胞术进行细胞周期分析.应用罗丹明（Rh123）排出试验定测P-gp功能.采用免疫组织化学方法检测多药耐药蛋白（MDR1）、多药耐药相关蛋白（MRP1）的表达情况.结果：Ec9706/P-1细胞与Ec9706细胞镜下形态无明显差别,体积稍大;Ec9706/P-1生长缓慢（P＜0.05）,倍增时间延长（P＜0.05）,大部分肿瘤细胞集中在G0＋G1期（P＜0.05）;MTT结果显示Ec9706/P-1对PIX的耐药指数为6.68,具有紫杉醇耐药性;Rh123排出试验显示Ec9706/P-1细胞平均荧光强度显著降低;免疫细胞化学方法检测发现Ec9706/P-1多药耐药蛋白（MDR1）表达明显升高（P＜0.05）,多药耐药相关蛋白（MRP1）表达无统计学意义（P＞0.05）.结论：Ec9706/P-1具有明确的耐药性,但耐药蛋白表达不一致,耐药机制复杂,可用于食管癌临床治疗的基础研究.     

人端粒酶hTRT在癌旁黏膜上皮和食管鳞癌组织中的表达及其意义

目的 研究食管黏膜鳞状上皮和食管鳞癌组织中端粒酶hTRT的表达，并探讨其与食管癌发生、发展的关系。方法 应用免疫组化S—P法，观察10例正常食管黏膜、35例癌旁食管黏膜上皮，120例食管癌组织微阵列中端粒酶hTRT的表达情况。结果 正常食管黏膜鳞状上皮细胞末见端粒酶hTRT阳性表达，癌旁黏膜上皮和癌组织端粒酶hTRT阳性表达串分别为94．3％和81．7％。癌组织hTRT的表达与癌组织的组织学分级、浸润深度和淋巴结转移无关(P＞0．05)。结论 端粒酶hTRT的激活表达与食管黏膜上皮的恶性转化密切相关，端粒酶hTRT的重新激活可能在食管癌的组织发生中起关键性作用。     

沉默stathmin基因增强食管鳞癌细胞对紫杉醇敏感度的实验

目的 探讨沉默stathmin基因后,食管鳞癌细胞对紫杉醇敏感度的改变。方法 利用RNA干扰技术特异性沉默食管鳞癌KYSE150细胞的stathmin基因;实时定量PCR及蛋白质印迹法检测stathmin表达变化;CCK-8检测紫杉醇敏感度变化;流式细胞仪检测细胞周期变化;细胞分裂指数实验观察细胞有丝分裂改变。结果 特异性转染后,stathmin基因mRNA 和蛋白表达均受抑制。沉默stathmin基因后,KYSE150细胞对紫杉醇敏感度增强191.4倍;紫杉醇对特异性转染和未转染的KYSE150细胞半数抑制浓度分别为0.018 nM和3.445 nM。经紫杉醇（0.01 nM）处理72 h后,相比于未转染细胞,特异性转染的KYSE150细胞出现明显G₂/M周期阻滞[（55.4±6.2%）vs.（19.1±2.7%）, P=0.000], 有丝分裂指数减少[（5.6±0.8%）vs.（13.5±3.7%）, P=0.000]。结论 stathmin基因沉默通过细胞周期G₂/M阻滞增强食管鳞癌细胞对紫杉醇的化疗敏感度。     

Expression and Clinical Significance of REPS2 in Human Esophageal Squamous Cell Carcinoma

Objective: REPS2 plays important roles in inhibiting cell proliferation, migration and in inducing apoptosisof cancer cells, now being identified as a useful biomarker for favorable prognosis in prostate and breast cancers.The purpose of this study was to assess REPS2 expression and to explore its role in esophageal squamous cellcarcinoma (ESCC). Methods: Protein expression of REPS2 in ESCCs and adjacent non-cancerous tissues from 120patients was analyzed by immunohistochemistry and correlated with clinicopathological parameters and patientoutcome. Additionally, thirty paired ESCC tissues and four ESCC cell lines and one normal human esophagealepithelial cell line were evaluated for REPS2 mRNA and protein expression levels by quantitative RT-PCR andWestern blotting. Results: REPS2 mRNA and protein expression levels were down-regulated in ESCC tissuesand cell lines. Low protein levels were significantly associated with primary tumour, TNM stage, lymph nodemetastasis and recurrence (all, P < 0.05). Survival analysis demonstrated that decreased REPS2 expression wassignificantly associated with shorter overall survival and disease-free survival (both, P < 0.001), especially inearly stage ESCC patients. When REPS2 expression and lymph node metastasis status were combined, patientswith low REPS2 expression/lymph node (+) had both poorer overall and disease-free survival than others (both,P < 0.001). Cox multivariate regression analysis further revealed REPS2 to be an independent prognostic factorfor ESCC patients. Conclusions: Our findings demonstrate that downregulation of REPS2 may contribute tomalignant progression of ESCC and represent a novel prognostic marker and a potential therapeutic target forESCC patients.     

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。     

Surgical Treatment for Early Esophageal Squamous Cell Carcinoma

More studies are needed to clarify treatments and prognosis of early esophageal squamous cell carcinoma(ESCC). This retrospective study was designed to review the outcome of surgical treatment for early ESCC,evaluate the results of a left thoracotomy for selected patients with early ESCC, and identify factors affectinglymph node metastases and survival. The clinicopathological data of 228 patients with early ESCC who underwenttransthoracic esophagectomy with lymphadenectomy without preoperative adjuvant treatment were reviewed.The χ2 test or Fisher’s exact test were used to detect factors related to lymph node metastasis. Univariate andmultivariate analyses were performed to identify prognostic factors. There were 152 males and 76 females witha median age of 55 years. Two hundred and eight patients underwent a left thoracotomy, and the remaining20 patients with lymph nodes in the upper mediastinum more than 5 mm in short-axis diameter by computedtomography scan underwent a right thoracotomy. No lymph node metastasis was found in the 18 patients withcarcinoma in situ, while lymph node metastases were detected in 1.6% (1/62) of patients with mucosal tumoursand 18.2% (27/148) of patients with submucosal tumours. Only 7 patients showed upper mediastinal lymph nodemetastases in the follow-up. The 5- and 10-year overall survival rates were 81.4% and 70.1%, respectively. Onlyhistologic grade (P<0.001) and pT category (P=0.001) significantly correlated with the presence of lymph nodemetastases. In multivariate analysis, only histologic grade (P=0.026) and pT category (P=0.008) were independentprognostic factors. A left thoracotomy is acceptable for selected patients with early ESCC. Histologic grade andpT category affected the presence of lymph node metastases and were independent prognostic factors for earlyESCC.     

Prognostic Significance of Desmoglein 2 and Desmoglein 3 in Esophageal Squamous Cell Carcinoma

Objective: Desmogleins (DSGs) are major members among the desmosomal cadherins critically involved incell-cell adhesion and the maintenance of normal tissue architecture in epithelia. Reports exploring links of DSGfamily member expression with cancers are few and vary. The aim of this study was to investigate the ratio ofDSG2 and DSG3 mRNA expression in esophageal squamous cell carcinoma (ESCC) tissue to normal tissue (T/Nratio) and evaluate correlations with clinical parameters. Methods: The mRNA expression of DSGs, as well asγ-catenin and desmoplakin, was detected by real-time quantitative RT-PCR in 85 cases of ESCC tissue specimens.Results: The expression level of DSG3 mRNA was significantly higher than that of DSG2 in ESCC specimens (p= 0.000). DSG3 mRNA expression highly correlated with histological grade (p = 0.009), whereas that of DSG2did not significantly relate to any clinicopathologic parameter. Kaplan-Meier survival analysis showed that onlyDSG3 expression had an impact on the survival curve, with negative DSG3 expression indicating worse survival(p = 0.038). Multivariate Cox regression analysis demonstrated DSG3 to be an independent prognostic factorfor survival. Furthermore, correlation analysis demonstrated the mRNA level of DSG3 to highly correlate withthose of γ-catenin and desmoplakin in ESCC samples (p=0.000), implying that the expression of desmosomalcomponents might be regulated by the same upstream regulatory molecules. Conclusions: Our findings suggestthat DSG3 may be involved in the progression of ESCC and serve as a prognostic marker, while expression ofDSG2 cannot be used as a predictor of ESCC patient outcome.     

人卵巢癌裸鼠移植腹水瘤模型的建立

建立人卵巢癌裸鼠移植腹水瘤模型,为研究人卵巢癌发病机制,生物学特性以及转移方式提供工具。方法将１例人卵巢癌的腹水直接接种于裸鼠腹腔,观察裸鼠腹水形以及肿瘤生长和转移情况,做腹水细胞学计数、涂片以及染色体和肿瘤标志物的检查,肿瘤和器官作病理学检查。结果成功建成１个人卵巢癌裸鼠移植腹水瘤模型,并已传至第１８代,传代移植成功率１００．０％,组织学和超微结构结构形态证实,保持了原人卵巢癌特征,具有人类肿瘤