外周血单个核细胞Th类细胞因子表达与食管鳞状细胞癌的相关研究

目的 :探讨外周血单个核细胞 (PBMC)Th1 、Th2 类细胞因子的表达与食管鳞状细胞癌临床病理因素之间的相关性。方法 :采用双抗体酶联免疫吸附法 (ELISA)检测 39例食管鳞状细胞癌患者和 2 0例健康对照者外周血单个核细胞Th1 (IL 2、INF γ)、Th2 (IL 4、IL 6、IL 1 0 )类细胞因子的含量水平 ,并与食管鳞状细胞癌病理因素之间进行相关分析。结果 :食管鳞状细胞癌患者PBMC中Th1 类细胞因子IL 2、INF γ的表达明显低于健康对照组 (P <0 .0 5) ,Th2 类细胞因子IL 6的表达明显高于对照组 (P <0 .0 5) ,存在Th1 /Th2 漂移现象 ;且发现TNM分期越晚 ,Th1 类细胞因子IL 2的表达越弱 ,而IL 6表达越强 ,存在显著性差异 (P <0 .0 5)。Th1 /Th2 漂移现象与肿瘤分化程度无明显相关性。结论 :食管鳞状细胞癌患者PBMC存在某些Th2 类细胞因子表达优势 ,而Th1 类细胞因子表达减弱 ;Th1 /Th2 漂移现象与肿瘤的TNM分期有关     

Kiss-1在食管鳞状细胞癌组织中的表达及临床病理意义

目的:观察Kiss-1蛋白及mRNA在食管鳞状细胞癌组织中的表达及其临床病理意义.方法:采用免疫组织化学SP法、原位杂交和逆转录聚合酶链反应(RT-PCR)技术,联合检测了62例食管鳞状细胞癌、31例癌旁不典型增生及62例正常食管组织中Kiss-1蛋白和mRNA的表达,并探讨其临床病理意义.结果:在食管鳞状细胞癌组织、癌旁不典型增生及正常食管组织中,Kiss-1蛋白的阳性表达率分别为56.5%、67.7%、90.3%(P<0.05);用原位杂交检测Kiss-1 mRNA阳性表达率分别为51.6%、74.2%、95.2%(P<0.05);用RT-PCR技术检测Kiss-1 mRNA阳性表达率分别为54.8%、71.0%、88.7%(P<0.05);食管鳞状细胞癌组织中Kiss-1蛋白及mRNA的低表达与淋巴结转移密切相关(P<0.05),而与患者的性别、年龄、肿瘤的组织学分级无关(P>0.05);Kiss-1蛋白的低表达与浸润深度有关(P<0.05);在食管鳞状细胞癌组织中,Kiss-1蛋白及mRNA的表达呈正相关(P<0.05),用原位杂交及RT-PCR技术对Kiss-1 mRNA表达情况的检测结果也呈正相关(P<0.05).结论:Kiss-1基因的表达降低或缺失和食管鳞状细胞癌的发生发展及转移有关,有望成为食管鳞状细胞癌早期诊断和预后判断的重要指标之一.     

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.     

中国河南林州市食管鳞状细胞癌组织中Ras蛋白的表达

癌基因ｒａｓ蛋白在细胞的生长和分化中起重要的作用,其编码的蛋白分子量约为Ｍｒ２１０００,称Ｐ２１蛋白,Ｐ２１蛋白位于细胞浆膜内面．具有与鸟苷酸结合的能力和ＧＴＰ酶的活性,参与膜的信号传递［１］．癌基因ｒａｓ突变,特别是第１２,１３和６１密码子部位的点突变可使细胞发生恶性转化［２］．消化道肿瘤,尤其是结肠癌,有较高的ｒａｓ基因突变发生率［１,２］,但是有关食管癌和ｒａｓ基因变化的关系的研究报道不尽一致［３,４］．我们通过对我国食管癌高发区食管癌组织中癌基因ｒａｓ蛋白的研究,进一步了解不同地区食管癌…     

食管鳞状细胞癌组织中基因表达差异的初步分析

目的 应用寡核苷酸芯片技术分析食管鳞状细胞癌、癌旁、正常组织中基因表达差异,探讨与食管鳞状细胞癌发生、发展相关的基因变化.方法 Trizol一步法抽提食管癌、癌旁、正常组织中总RNA,纯化后逆转录为cRNA,经荧光标记、纯化,与Agilent寡核苷酸芯片(21 074探针)杂交,应用Agilent扫描仪获取图像,特征提取软件定量分析处理.挑选明显差异表达的基因进行实时荧光定量逆转录-PCR、免疫组化、免疫印迹验证.结果 ①经寡核苷酸芯片技术筛选差异表达基因的结果显示:上调38个、下调61个.②实时荧光定量逆转录PCR验证提示:差异较大的上调基因有CTHRC1、INHBA、SPP1、LUM、HRK,其中CTHRC1差异最为显著.③免疫组化:CTHRC1在食管鳞癌组织中有增强表达,表达率为56.5%(26/46).在淋巴结转移组中的阳性表达率显著高于无淋巴结转移组(P<0.05).④免疫印迹:在食管癌细胞株TE-13及Eca-109中均检测到CTHRC1的表达.结论 CTHRC1可能是食管鳞癌最为重要的生物分子之一.     

食管鳞状细胞癌VEGF的表达及意义

肿瘤内新生血管可提供肿瘤生长必需的养料和转运其代谢产物,同时为肿瘤发生血性转移提供通道.新近研究表明,肿瘤血管形成是由多种生物活性因子调节控制的,其中血管内皮细胞生长因子(VEGF)在肿瘤新生血管形成中的作用尤为重要.     

替吉奥治疗老年晚期食管鳞状细胞癌的临床效果

目的探讨替吉奥(S-1)单药在老年晚期食管鳞状细胞癌一线治疗的临床疗效及不良反应。方法搜集80例经病理确诊的老年晚期食管鳞状细胞癌患者,试验组41例接受S-1胶囊口服,体表面积<1.25 m2:S-1 80 mg/d,1.25~1.50 m2:S-1 100mg/d,>1.50 m2:S-1 120 mg/d,第1~14天分2次口服;每21 d为1个周期,至少完成2个周期,同时接受最佳的支持治疗。对照组39例只接受最佳的支持治疗。根据RECIST标准评价其近期疗效及NCI毒性评价标准评价不良反应。比较两组有效率(RR)、疾病控制率(DCR)、无进展生存时间(PFS)和总生存时间(OS)。结果试验组RR为24%,DCR为53%。两组PFS、OS差异有统计学意义(P<0.05)。试验组不良反应主要为消化道及血液学毒性,白细胞减少发生率显著高于对照组(P<0.05),两组其余血液学毒性及非血液学毒性差异无统计学意义(P>0.05)。结论 S-1一线治疗老年晚期食管鳞状细胞癌有一定的疗效,不良反应可耐受。     

食管鳞状细胞癌患者单个核细胞Th类细胞因子表达的临床意义

为探讨外周血单个核细胞(PBMC)Th_1、Th_2类细胞因子的表达与食管鳞状细胞癌病理因素之间的关系,采用双抗体酶联免疫吸附法(ELISA)检测39例食管鳞状细胞癌患者外周血Th_1、Th_2类细胞因子的含量水平,同时以20例正常人作为健康对照。结果显示,食管鳞状细胞癌患者PBMC中IL-2、INF-γ表达明显低于对照组(P<0.05),IL-6表达明显高于对照组(P<0.05),存在Th_1/Th_2漂移现象;且TNM分期越晚,Th_1类细胞因子IL-2表达越弱,IL-6表达越强,存在显著差异性(P<0.05)。Th_1/Th_2漂移现象与肿瘤分化程度无明显相关性。提示食管鳞状细胞癌患者PBMC存在Th_2细胞优势,而Th_1类细胞因子表达减弱;Th_1/Th_2漂移现象与肿瘤的TNM分期有关。     

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     

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.     

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。     

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.     

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.     

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.     

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.