【作者】 张聪；

【导师】 王宝成；

【作者基本信息】 泰山医学院 ， 肿瘤学， 2010， 硕士

【摘要】 目的分析人食管鳞癌(ESCC)中HuR.VEGF-C的表达与患者临床病理特征及预后的关系,为筛选食管鳞癌新的治疗靶点提供实验依据,明确能否作为筛选早期食管鳞癌及淋巴结转移的生物学标记物。方法1、应用免疫组织化学染色(S-P法)检测58例人食管鳞癌组织和20例癌旁正常组织中HuR.VEGF-C的表达,分析二者之间的相关性及其与临床病理特征的关系。2、采用Kaplan-Meier法和Cox比例风险回归模型,分析HuR.VEGF-C的表达与术后生存的关系。3、应用ELISA法检测58例食管鳞癌患者及20例正常健康志愿者血清中VEGF-C水平,分析其与临床病理参数的关系。结果1、58例ESCC组织中,HuR胞浆阳性表达率、HuR胞核阳性表达率、VEGF-C阳性表达率分别为46.55%(27/58)、75.86%(44/58)和68.97%(40/58),与正常对照组的0%、3.45%和15.00%比较差异均有显著的统计学意义(P<0.01)。胞浆HuR.VEGF-C的表达与ESCC患者的淋巴结转移、肿瘤浸润深度和TNM分期均有显著的相关性(P<0.05)。胞浆HuR表达还与肿瘤大小有关(P<0.05)。胞核HuR表达与ESCC患者所有的临床病理特征均无相关性；2、食管鳞癌中胞浆HuR.VEGF-C的表达水平间存在明显的等级正相关性(r=0.351,P<0.05)；3、胞浆HuR阳性表达组的5年生存率明显低于阴性表达组(P<0.05)；胞核HuR阳性表达组的5年生存率虽然低于阴性表达组,但无统计学意义；VEGF-C阳性表达组的5年生存率明显低于阴性表达组(P<0.05)；胞浆HuR、VEGF-C双阳性表达组的5年生存率明显低,于双阴性表达组(P<0.05)；Cox回归模型的多变量分析结果显示,食管鳞癌患者预后的独立危险因素包括胞浆HuR表达(RR=2.344：95%C1,1.140-4.820；P=0.021)、VEGF-C表达(RR=2.565：95%Cl,1.087-6.052；P=0.032)以及淋巴结的转移(RR=2.233；95%Cl,1.068-4.668；P=0.033),而与胞核HuR的表达及其他临床病理特征均无相关性(P>0.05)4、58例ESCC患者与20例正常健康志愿者者血青VEGF-C表达水平分别为(643.58±144.56)pg/ml和(542.74±117.55)pg/ml,二者比较,差异具有显著的统计学意义(P<0.01)。Ⅲ、Ⅳ期患者血清VEGF-C的浓度(665.51±141.20)pg/ml和Ⅰ、Ⅱ期患者血清VEGF-C的浓度(574.66±137.61)pg/ml之间存在统计学上显著的差异(P<0.05)；有淋巴结转移的ESCC患者血清VEGF-C水平(685.51±142.52)pg/ml显著高于无淋巴结转移的ESCC患者的血.清VEGF-C水平(601.65±136.34)pg/ml(P<0.05)；与肿瘤浸润未超过外膜(T1+T2)的患者血清VEGF-C的水平为(576.01±140.96)pg/ml相比,肿瘤浸润超过外膜(T3+T4)的患者血清VEGF-C的水平(669.33士139.00)pg/ml显著升高。上述结果提示ESCC患者血清中VEGF-C表达水平与TNM分期(P<0.05)、淋巴结转移(P<0.05)以及肿瘤浸润深度(P<0.05)均有显著相关性。结论ESCC组织中HuR、VEGF-C表达和外周血VEGF-C水平与肿瘤进展、淋巴结转移密切相关,是ESCC的不良预后因子,可作为潜在的抗肿瘤靶点。更多还原

【Abstract】 ObjectiveStudy the expression of HuR and VEGF-C in Esophageal Squamous Cell Carcinoma (ESCC), and correlate results with patients’clinicopathological parameters and survival for providing experimental basis to screen new target for the treatment of ESCC, assessing their usefulness as biological markers for determining early ESCC and lymph node metastasis.Methods1. The expression of HuR and VEGF-C in 58 ESCC tissues and 20 control paracancerous normal tissues were detected by immunohistochemistry staining S-P assay. The correlation between the expression of HuR and VEGF-C and the clinicalpathological parameters and the relationship between the expression of HuR and VEGF-C were analyzed.2. The correlation between HuR and VEGF-C expression and patient outcome was determined by Kaplan-Meier method and Cox proportional hazards regression model.3. Serum VEGF-C levels in 58 patients with ESCC and 20 healthy people were measured by ELISA assay, and the relationships between VEGF-C and different clinicopathological factors was analyzed.Results1. In 58 ESCC tissues, positive rate of cytoplasmic HuR, nuclear HuR and VEGF-C were 46.55%(27/58),75.86%(44/58) and 68.97%(40/58), respectively. Compared With positive rate of cytoplasmic HuR, nuclear HuR and VEGF-C in normal Esophagal tissues,0%(0/20),10.00%(2/20)and 15.00%(3/20) respective-ely,the difference had a statistic significance (P<0.01). Both the expression of cytoplasmic HuR protein and VEGF-C protein were positively correlated with lymph node metastasis, depth of tumor invasion and TNM stages (P<0.05). In addition, the expression of cytoplasmic HuR protein was positively correlated with tumor size. However, the expression of nuclear HuR protein was not correlated with any clinicopathological factors.2. There was an obvious positive correlation between the expression of cytoplasmic HuR and VEGF-C (r=0.351, P<0.05).3. The follow-up visit in the patients showed that five-year survival rate was greatly lower in the positive expression of the cytoplasmic HuR group than that in the negative group (P<0.05). Although the positive expression of nuclear HuR group had a lower 5 year survival rate than the negative group, there is no statistic significance. Meanwhile, the positive expression of VEGF-C group had a lower 5 year survival rate than the negative group obviously (P<0.05). The 5 year survival rate in the cytoplasmic HuR and VEGF-C double positive group was lower than the double negative group obviously. In multivariate analysis using the Cox regression model, the independent prognostic parameters for ESCC patients included cytoplasmic HuR express(RR=2.344; 95% Cl,1.140-4.820;P=0.021), VEGF-C express (RR=2.565; 95% Cl,1.087-6.052;P=0.032) and lymph node metastasis (RR=2.233; 95% Cl,1.068-4.668; P=0.033), had no correlation with nuclear HuR express and other clinicopathological characteristics (?>0.05)。4. We found that the serum VEGF-C level in 58 ESCC patients and 20 healthy controls was (643.58±144.56) pg/ml, (542.74±117.55) pg/ml, respectiv-ely,indicated an obvious statistic difference (P<0.01). The concentrations of VEGF-C were statistically significantly different in stage III and stage IV ESCC patients (665.51±141.20) pg/ml and stage I and stageⅡESCC patients (574.66±137.61) pg/ml (P<0.05). ESCC patients with lymph node metastasis had a higher VEGF-C level (685.51±142.52) pg/ml than those without lymph node metastasis (601.65±136.34) pg/ml (P<0.05).Compared with serum VEGF-C concentration (576.01±140.96) pg/m in patients whose depth of tumor invasion did not overpass serous membrane (T1+T2), serum VEGF-C concentration (576.01±140.96) pg/m in patients whose depth of tumor invasion overpassed serous membrane (T3+T4) elevated notably. Above-mentioned results suggested that Serum VEGF-C level in ESCC patients was closely correlated with TNM stage (?<0.05), lymph node metastasis (P<0.05) and depth of tumor invasion (P<0.05). ConclusionBoth the expression of HuR and VEGF-C in ESCC tissues and VEGF-C level in peripheral blood are closely correlated with malignant tumor progress and lymph nodes metastasis.Therefore, they are considered as poor prognosis markers of ESCC, become potential anti-tumor therapeutic targets.更多还原