【作者】 赵一鸣；

【导师】 王鲁；

【作者基本信息】 复旦大学 ， 外科学， 2008， 硕士

【摘要】 研究目的:肝细胞癌(hepatocellular carcinoma,HCC)是典型的多血管肿瘤,新生血管生成与肝癌预后有关。本实验在前期证实通过抑制血管生长因子表达阻断血管生成进而抗肝细胞癌转移复发和肿瘤生长的基础上进一步检测肝细胞癌患者术前血浆中多种血管生长因子,包括巨噬细胞移动抑制因子(macrophagemigration inhibitory factor,MIF)、血管内皮生长因子(vascular endothelialgrowth factor,VEGF)、白细胞介素-8(interleukin-8,IL-8)的表达和癌组织中微血管密度(microvessel density,MVD)。分析血管生成因子相互之间及血管生成、肿瘤临床病理特征及预后的关系。为临床提供潜在的预测肝癌预后的指标和提高肝癌疗效的作用靶点。对象和方法:收集中山医院肝外科2005年1月—2005年12月期间初次确诊入院行根治性切除术的HCC患者44例和同期确诊为肝良性占位患者20例(包括肝血管瘤8例,肝囊肿10例,肝局灶性结节性增生2例)。应用双抗夹心酶联免疫吸附实验(enzyme-linked immunosorbentassay,ELISA)测定血浆MIF、VEGF、IL-8浓度。免疫组化法检测HCC组织MVD。分析血浆MIF水平与组织MVD及血浆VEGF水平的关系,探讨血浆MIF水平与HCC血管生成的关系。收集临床资料(患者性别、年龄、HBsAg)、病理学资料(肿瘤大小、数目、包膜完整性、血管侵犯、肿瘤分化程度、TNM分期)以及随访资料(转移复发时间、死亡时间及原因、生存状态),分析血浆MIF水平和临床病理特征及生存预后的关系。结果:44例HCC患者血浆MIF中位浓度为9.09 ng/ml(四分位距,4.54-20.45ng/ml),血浆VEGF中位浓度76.13 pg/ml(四分位距,22.42-148.01 pg/ml)。22例肝良性占位患者血浆MIF中位浓度4.80 ng/ml(四分位距,2.39-6.80ng/ml),血浆VEGF中位浓度25.40 pg/ml(四分位距,20.16-37.58 pg/ml),HCC患者血浆MIF浓度高于肝良性占位患者,差异有统计学意义(P=0.002)。HCC患者血浆MIF、VEGF浓度呈正相关关系(r=0.547,P＜0.001)。HCC组织MVD中位数为33.7/HPF(四分位距,20.3-41.9/HPF),比较HCC患者血浆MIF浓度和MVD后发现血浆HCC患者血浆MIF浓度和组织MVD呈正相关关系(r=0.747,P＜0.001)。血浆MIF浓度和肿瘤数目(P＜0.001)、大小(P=0.039)、包膜完整性(P=0.044)、血管侵犯(P=0.041)、临床TNM分期(P＜0.001)有关,多发、直径＞5cm、无包膜或包膜不完整、有血管侵犯肿瘤患者的血浆MIF浓度高于单法、直径≤5cm、有完整包膜、无血管侵犯肿瘤患者。低MIF表达(＜9.09 ng/ml)患者的总体生存时间(中位总体生存时间,26.9个月vs20.3个月;P=0.0069)和无瘤生存时间(中位生存时间,24.5月个vs12.3个月;P=0.0023)明显高于高MIF表达(＞9.09 ng/ml)患者。应用COX回归模型进行单因素和多因素分析结果均表明血浆MIF水平是影响肝癌患者术后总体生存和无瘤生存的独立危险因素。结论:HCC患者血浆MIF水平和血浆VEGF水平及肿瘤MVD呈正相关,MIF可能通过刺激VEGF分泌影响肿瘤血管生成。血浆MIF水平与HCC患者临床病理特征及术后复发和生存预后密切相关,有望成为应用于临床的肝癌侵袭性的生物学指标和患者预后的独立预测指标。更多还原

【Abstract】 Objective:We measured the plasma levels of MIF and microvessel density(MVD) of tumor tissue in 44 patients with hepatocellular carcinoma(HCC) after curative resection to investigate its relationship with HCC angiogenesis and roles in predicting prognosis of the overall survival and disease-free survival.Material and methods:Forty-four consecutive Chinese patients who underwent curative resection of HCC in the Department of Hepatic Surgery(Zhongshan Hospital,Fudan University) between January 2005 and December 2005 were recruited randomly into this study after informed consent.As negative control,twenty patients with liver benign lesions (including 8 patients with haemangioma,2 patients with focal nodular hyperplasia,10 patients with cyst) were enlisted as well.Enzyme-linked immunosorbentassay (ELISA) was used to detect the concentrations of MIF,vascular endothelial growth factor(VEGF) and interleukin-8(IL-8) in preoperative plasma of 44 patients with curative resection of HCC and 20 negative subjects.Immunohistochemistry was used to measure microvessel density(MVD) stained by anti-CD34 antibody.The relationship between MIF and angiogenesis was evaluated by comparing plasma MIF level and MVD of HCC.The relationship between plasma MIF level of HCC with clinicopathological feature(gender,age,HBsAg status,tumor size,tumor number, tumor capsule,venous invasion,tumor differentiation,TNM stage),disease-free survival and overall survival of patients after curative resection were analyzed.The prognosis value of plasma MIF level was evaluated by survival analysis and Cox regression. Results:The preoperative plasma MIF levels of 44 patients with resection of HCC ranged from 1.42 to 78.8 ng/ml,with a median of 9.09 ng/ml(interquartile range, 4.54-20.45ng/ml).The average level of plasma MIF in these 44 patients with HCC was significantly higher than that of healthy controls(median,4.80 ng/ml; interquartile range,2.39-6.80ng/ml;P=0.002).The plasma VEGF levels of HCC patients ranged from 10.57 to 654.78 pg/ml,with a median of 76.13 pg/ml (interquartile range,22.42-148.01 pg/ml),the average level of plasma VEGF in HCC patients was significantly higher than that of healthy control(median,25.40 pg/ml; interquartile range,20.16-37.58 pg/ml;P=0.002).To our interest,all but one plasma samples were undetectable for IL-8.Analyzed by the Pearson correlation coefficient, plasma MIF levels correlated positively with VEGF(r=0.547,P＜0.001).The intratumoral MVD ranged from 20.3/HPF to 96.2/HPF,with a median of 33.7/HPF (interquartile range,28.0-41.9/HPF).The MVD was significantly higher in tumors with high plasma MIF level than in tumors with low plasma MIF level.Plasma MIF level positively correlated with MVD(r=0.747,P＜0.001).There was no significant correlation between plasma MIF concentration and patients’ gender(P=0.166),age (P=0.558),hepatitis B surface antigen status(P=0.370) and Edmonson grade (P=0.421).whereas high plasma MIF levels were significantly associated with large tumors＞5cm(P=0.039),multiple tumor(P＜0.001),absence or unintegrity of tumor capsule(P=0.044),presence of venous invasion(P=0.041) and an advanced disease stage(P＜0.001).The plasma MIF levels in patients with stageⅢorⅣdisease were significantly higher than those of patients with stageⅠorⅡdisease.The median overall survival of patients in low MIF group was significantly better than that of patients in high MIF group(median overall survival,26.9 months versus 20.3 months; P=0.0069).The median disease-free survival of patients in low MIF group was also significantly better than that patients in high MIF group(median disease-free survival, 24.5 months versus 12.3 months;P=0.0023).A high plasma MIF level was significantly associated with a poorer overall survival(P=0.0069) as well as disease-free survival(P=0.0023),and was one of the leading independent prognostic factors for both overall survival(hazard ratio=3.279;95%confidence interval 2.095-8.821;P=0.02) and disease-free survival(hazard ratio--2.256;95%confidence interval 2.100-7.652;P=0.004) in multivariate COX model analyses.Conclusions:The plasma MIF level of HCC positively correlated with MVD and plasma VEGF level,it is maybe enhancing VEGF level by which MIF stimulates angiogenesis of HCC.There are significant relationships between plasma MIF level and clinicopathological features,disease-free survival and overall survival,so the evaluation of preoperative plasma MIF level may be helpful to predict the recurrence and prognosis of HCC patients in advance.更多还原