【作者】 林玲；

【导师】 周建英；

【作者基本信息】 浙江大学 ， 内科学， 2009， 硕士

【摘要】 背景与目的:肺癌是我国最为常见的恶性肿瘤,发病机制迄今未明。目前研究发现人β防御素-3(beta-defensin-3,HBD-3)是种内源性抗菌肽,具有趋化作用,在人肝癌细胞,人外阴肿瘤,人口腔鳞状上皮细胞癌组织中均有HBD-3的mRNA和蛋白水平的高表达。亦有研究发现HBD-3在肺癌中的mRNA表达量明显升高,但在蛋白水平的研究尚未见报道。HBD-3除具有强大的直接抗微生物效应外,还参与适应性免疫调节,并可能与肿瘤免疫有关,HBD-3在肺癌免疫中的作用有待于进一步研究。本研究探讨了β防御素-3表达与树突状细胞浸润(Dendritic cells,DC)和非小细胞肺癌免疫的关系。方法:收集2007年1月至2009年1月本院病理组织库采集的60例肺癌病例外科手术切除标本。所有病例术前均未接受任何抗肿瘤治疗(化疗/放疗),包括鳞癌31例,腺癌25例,支气管肺泡癌4例,其中Ⅰ期患者12例,Ⅱ期28例,Ⅲ期20例,每个病例均包括肺癌组织、正常组织两部分,(1)采用免疫组化方法(EnVision法)测定上述两部分组织HBD-3表达的阳性率,同时对其在不同病理类型及不同分期肺癌组织表达的阳性率进行比较分析。(2)以S100蛋白作为DC特异性标记蛋白,采用EnVision法测定S100阳性细胞在肺癌组织、正常组织浸润程度,同时分析HBD-3与DC在肺癌组织中表达的相关性。结果:1.在非小细胞肺癌组织及周边正常组织细胞胞质内均有不同程度的HBD-3阳性的表达。其中,肺癌组织内HBD-3表达阳性率最高为70.0%,正常组织为20.0%,比较肺癌组织与正常组织P值分为0.000,有统计学上显著差异。2.在不同病理类型中的肺癌组织内有不同程度HBD-3阳性的表达,以鳞癌组织内表达密度最高,鳞癌与腺癌分别为90.3%、48.0%,两者表达存在显著性差异(P＜0.001)。3.HBD-3在不同的临床阶段(Ⅰ期、Ⅱ期、Ⅲ期)的肺癌组织中表达相似,阳性率分别为50.0%、75.0%、75.0%,两两比较,Ⅰ期与Ⅱ期、Ⅰ期与Ⅲ期、Ⅱ期与Ⅲ期,P值分别为0.122、0.149、1.000,无显著差异。4.肺癌组织中S100阳性率明显高于正常组织,两者的阳性率分别为78.3%,16.7%,比较肺癌组织与正常组织有显著差异,P值分为0.000。5.采用spearman等级相关分析HBD-3与S100阳性的树突状细胞在肺癌组织及正常组织分布的关系,二者呈显著正相关(P＜0.05),提示HBD-3在肺癌组织的表达含量与DC的浸润有关。结论:在非小细胞肺癌组织内存在高水平的HBD-3蛋白表达,其表达程度与癌组织DC的浸润有关,提示HBD-3可能通过趋化未成熟的DC而在机体抗肿瘤免疫中发挥作用。更多还原

【Abstract】 Background and Objective:Lung cancer is one of the most common malignant tumors with poor prognosis.Although the mechanism remains unclear,increasing evidence show that human beta-defensin 3(HBD-3),regarded as one kind of endogenous antimicrobial peptide and chemokine,is high expressed on human liver cancer,human vulva tumor and human oral squamous cell carcinoma on mRNA level and protein level.In lung cancer the mRNA expression of HBD-3 is high,while the protein expression is not known yet.HBD-3 directly takes part in reactions of anti-microorganism and adjustability of immunity modulation,may also affects the immunologic reaction in tumors.Thus,further studies are required to determine the function of HBD-3 in lung cancer.In this study we will focus on the the relationship between HBD-3,dendritic cells infiltrating and non-small cell lung cancer immunity.Methods:60 lung cancer tissues samples were collected by pathologic tissues center of Taizhou hospital from Jan 2007 to Jan 2009.All patients did not receive any chemotherapy or radiotherapy before surgeries.These samples included squamous cell carcinoma(31 cases),adenocarcinoma(25cases) and bronchoalveolar carcinoma(4 cases) while by TNM staging there were 12 cases at stageⅠ,28 cases at stageⅡand 20cases at stageⅢEach sample contained tumor tissue and normal tissue.We respectively detected(1) the positive expression rate of HBD-3 in these two parts by immunohistochemistry(EnVision),and then analyzed the data according to pathologic type and TNM stage;(2)the density of dendritic cells infiltrating in each sample by Envision using protein S-100 as DC-specific label marker,and then analyzed the correlation between HBD-3 expression and DC infiltration in the lung cancer tissues.Results:(1) The expression of HBD-3 in non-small cell lung cancer was enhanced significantly(P=0.000),compared to that in normal tissues.The positive expression rates of HBD-3 in tumor is 70.0%,while in normal tissues it is only 20.0%.(2) The expression of HBD-3 in squamous cell carcinoma was enhanced significantly,compared to that in adenocarcinoma(P＜0.001).The positive rate of HBD-3 expression in squamous cell carcinoma and adenocarcinoma was 90.3%and 48.0%,respectively.(3) There were no significant differences of HBD-3 expression in lung cancer tissues amongⅠ,ⅡandⅢTNM stage.(4) The positive rate of S100-labeled DCs in tumor issues is significantly increased,compared to the rate in normal tissue(P＜0.05).(5) There is significant correlation between HBD-3 expression and DC infiltration in the tumor tissues by using spearman correlation analysis (P＜0.05).Conclusion:The data shows that HBD-3 is high expressed in the non-small cell lung cancer tissues,and the expression level is positively correlated with DC infiltration,suggesting that HBD-3 may play an important role in anti-tumor immunity through aggregating immature DCs in tumor tissues.更多还原