【作者】 宋阳；

【导师】 王莉芬；

【作者基本信息】 大连医科大学 ， 病理学与病理生理学， 2008， 硕士

【摘要】 鼻咽癌是指发生于鼻咽顶部和侧壁的恶性肿瘤,在亚洲人口发生率较高,同时也是我国高发恶性肿瘤之一。发病以男性为主(男:女=2:1),好发年龄: 40-49岁,其次30-39岁和50-59岁,其发病年龄比其它常见的头颈部恶性肿瘤年轻,小儿科也较常见。目前认为鼻咽癌发生与遗传、病毒感染及环境因素等有关,尤其Epstein-Barr病毒(EBV)与鼻咽癌发生、发展密切相关。鼻咽癌的病理分型大多为分化型非角化性癌和未分化癌,临床治疗多首选放射治疗,在放疗期间配合化疗和/或中医中药治疗,放疗3个月仍有残余病灶可采用手术治疗。放疗虽然对早期鼻咽癌有效控制率达85%,但由于其可引起口腔黏膜干燥,听力丧失和中耳炎,牙关紧闭,放射线照射下视丘、颞叶、和脑下垂体的后遗症等并发症,影响了肿瘤患者的生存质量,因此限制了放疗在临床抗肿瘤治疗上的应用。随着肿瘤免疫学的发展,肿瘤的综合治疗逐渐成为主流,出现了EBV特异性过继免疫治疗及分子靶向治疗等新的治疗进展。其中过继性细胞免疫治疗是指向肿瘤患者转输具有抗肿瘤活性的免疫细胞,直接杀伤肿瘤或激发机体抗肿瘤的免疫效应,从而达到治疗肿瘤的目的。这与肿瘤的细胞免疫密切相关。随着免疫学的发展和对疾病机制认识的提高,人们越来越发现人类疾病的发生发展与其免疫状况密切相关,机体免疫状态低下则被认为是病毒致癌的重要因素,正是认识到这一点国内外多篇文献报道运用流式细胞技术检测恶性肿瘤患者的外周血中淋巴细胞亚群分布情况,发现恶性肿瘤患者外周血CD3~+细胞、CD4~+细胞明显减少, CD4~+ / CD8~+比值明显下降,提示恶性肿瘤患者细胞免疫功能减退,但有关鼻咽癌组织中淋巴细胞亚群分布研究较少。目前在鼻咽癌肿瘤内科治疗方面,环氧化酶-2(cyclooxygenase-2, COX-2)分子靶向治疗更是研究的热点。COX-2是花生四烯酸转化为前列腺素过程中的重要限速酶之一,它与细胞增殖与分化、肿瘤的浸润转移及血管生成密切相关。在正常生理状态下多数组织内检测不到COX-2,但在细胞因子和促肿瘤剂等因素的刺激下COX-2的表达可上调。近年来研究报道淋巴细胞分泌的细胞因子可上调COX-2的表达,但在鼻咽癌组织中COX-2的表达以及与淋巴细胞亚群的关系研究报道甚少。目的:了解鼻咽癌组织中淋巴亚群的特点、COX-2的表达情况以及淋巴细胞亚群与COX-2表达的相关性,探讨癌组织局部免疫和COX-2在鼻咽癌发生发展中的作用,为鼻咽癌的过继免疫治疗和COX-2分子靶向治疗提供的实验基础。材料与方法:收集大连医科大学附属二院2002年-2007年收集的存档鼻咽癌蜡块45例,28例为未分化型非角化性癌;17例为分化型非角化性癌。45例鼻咽癌中有32例有颈淋巴结转移。同时取33例鼻咽部慢性炎症病例用于对照。所以标本均重新切片、HE染色,光镜下观察,再次明确病理诊断。用免疫组化方法检测鼻咽癌与鼻咽部慢性炎症组织CD3~+,CD4~+, CD8~+, CD20~+及COX-2表达情况,并将两者加以对照,同时将COX-2检测结果与淋巴亚群表达情况进行相关性分析。最后应用SPSS13.0统计软件,用X2分析对实验结果进行统计学处理,同时运用Spearman判断其相关性,差异显著性标准为P<0.05。结果:1. CD3~+细胞表达率:鼻咽癌为53.34%,其中+ 13例(28.89%)、++7例(15.56%)、+++4例(8.89%)。慢性鼻咽炎阳性表达率为81.81%,其中+13(39.39%)、++5例(15.15%)、+++9例(27.27%)鼻咽癌CD3阳性表达率低于慢性鼻咽炎组,两组间差异具有统计学意义(χ2=8.96,P=0.030)。2. CD4~+细胞表达率:鼻咽癌为20. 00%,其中+ 9例(20. 00%),其他均未表达。慢性鼻咽炎阳性表达率为42.42%,其中+14(42.42%),其他均未表达。鼻咽癌CD4阳性表达率低于慢性鼻咽炎组,两组间差异具有统计学意义(χ~2=4.60,P=0.032)。3. CD8~+细胞表达率:鼻咽癌为77.77%,其中+ 24例(53.33%)、++ 9例(20.00%)、+++ 2例(4.44%)。慢性鼻咽炎阳性表达率为78.79%,其中+ 22(66.67%)、++ 4例(12.12%)、+++ 0例(0%)两组间阳性率差异无统计学意义(χ~2=3.51,P=0.319)。4. CD20~+细胞表达率:鼻咽癌为48.88%,其中+ 12例(24.44%)、++ 6例(13.33%)、+++ 5例(11.11%)。慢性鼻咽炎阳性表达率为63.63%,其中+ 10(30.30%)、++ 7例(21.21%)、+++ 4例(12.12%)。两组间阳性率差异无统计学意义(χ~2=1.89,P=0.595)。5. COX-2的表达率:鼻咽癌为86.67%,其中+ 20例(44.44%)、++12例(26.67%)、+++7例(15.56%)。慢性鼻咽炎阳性表达率为93.94%,其中+6(18.18%)、++11例(33.33%)、+++14例(42.42%)鼻咽癌组COX-2阳性表达率低于慢性鼻咽炎组(χ~2=10.31,P=0.016)。6.鼻咽癌组织中CD3、CD4、CD8、CD20分别与COX-2阳性表达率间进行Spearman相关性分析,其中CD4与COX-2的阳性表达率间有相关性(R=0.464,P=0.023), CD3、CD8、CD20与COX-2的阳性表达率间无相关性(R=0.245,P=0.249; R=-0.102,P=0.634; R=0.000,P=0.998)。结论:1.鼻咽癌组织中CD3+表达率较鼻咽炎组织减少,而两者的CD20+表达差异不明显,提示鼻咽癌患者鼻咽部组织确实存在免疫功能紊乱,主要是细胞免疫功能障碍。2.鼻咽癌组织中CD3+表达减少主要是CD4~+表达减少,而CD8~+表达变化不明显,提示CD4~+/ CD8~+比值降低使鼻咽部组织免疫功能受到抑制。3.鼻咽癌组织中CD4与COX-2阳性表达率正相关,提示CD4细胞可通过某种途径调节COX-2的表达。更多还原

【Abstract】 Nasopharyngeal carcinoma is a malignant tumor which derived from side and crest of nasopharyngeal; it has high mobility in Asia. It is also one of the highest mobility malignant tumors. Men is the main, the ordinary age is : 40-50; the second : 30-39 and 50-59. Now nasopharyngeal carcinoma is related to heredity、Epstein-Barr virus、environment, especially Epstein-Barr virus.In nasopharyngeal carcinoma, low differentiation squamous cell carcinomas can be seen usually. Radiotherapy is the first choice , during the radiotherapy,we can also chose chemotherapy and the Chinese medical science and traditional Chinese medical . We can adopt surgery if pathological changes are left after three month. Although the rate of effective controlling is 85%, but radiotherapy can lead the complication : the membrane of oval cavity dry, loss of hearing and secretors toasts media ; lingering effects of brain and the brainstem and vertebra. With the side effect of radiotherapy and chemotherapy,it affects living quality of patients. Radiotherapy is restricted in clinical. with the development of tumor immunitiology, Tumor composite treatment plays an important role. Here comes new progress for example adoptive cell therapy, another is target treatment. Adoptive cell therapy means, we can put activity immunity cell into the patient to kill the tumor. It is closed to cell immunity. With the development of immunology, people come to know that the disease is closed to immunity. Immunity plays an important role in cancer. Many articles have been reported with flow cell cytometry that CD3~+↓、CD4~+↓, CD4~+ / CD8~+↓obviously in malignant tumor. It points out that the function of cell immunity of malignant tumor goes down. This experiment We focus on the change of lymphocyte subpopulations in nasopharyngeal carcinoma, to learn the immunity answer to it. This is better to Adoptive cell therapy. It has positive clinical purpose.At present, the target treatment of COX-2 is a new hot focus. Cyclooxygenase-2 is one of important enzymes during cyclooxyge transform into prostate simple. It is associated with cell proliferation and differentiation, tumor infiltration and metastasis, angiogenesis. It can not be detected normally, but IL-2、TNF-αwhich CD4 secreted can up regulate the expression of COX-2.Our study detects the expression of T lymphocyte subpopulations and COX-2 in nasopharyngeal carcinoma and referent the results about COX-2 of group mates to analyze their relationship, which can provide us with available information for the target treatment of nasopharyngeal carcinoma.Objective: We adopt Immunohistochemistry method to check the expression of T lymphocyte subpopulations and COX-2 about nasopharyngeal carcinoma and the contrast. Explore the correlation of T lymphocyte subpopulations and COX-2 expression. It is important for further study the relationship between COX-2 and cell immunity, it provides available experiment for bases for adoptive cell therapy treatment and target treatment.Method: Nasopharyngeal carcinoma with clear pathological diagnosis 45 patients from The Second Affiliate of Dalian Medical University between 2002-2007.Barbarism cancer 27, scale 18, 32 lymph transfer, and 13 non-lymph transfer, nasopharyngeal with chronic nasopharyngitis 33 is on the contrast. Immunohistochemistry method was used to detect CD4~+, CD8~+, CD3~+, CD20~+ and COX-2 in canceration .Analyze the relationship between COX-2 and CD4~+, CD8~+, CD3~+,CD20~+.The experiment results were analyzed with X~2 test and Spearman by SPSS 13.0 statistic software, and P<0.05 was used as the criteria of marked differentia.Result:1.The positive rates of CD3+: the positive rate in nasopharyngeal carcinoma is 53.34%, including 13cases of +(28.89%),7 cases ++(15.56%) and 4 cases of +++(8.89%); the positive rate in nasopharyngitis is 81.82%, including 13cases of +(39.39%), 5 cases of ++(15.15%) and 9 cases of +++(27.27%).The positive rate in nasopharyngeal carcinoma significantly lower than that in nasopharyngitis(χ~2=8.96, P=0.030).2. The positive rates of CD4~+: the positive rate in nasopharyngeal carcinoma is 20%, including 9 cases of +(20%); the positive rate in nasopharyngitis is 42.42%, including 14cases of +(42.42%). The positive rate in nasopharyngeal carcinoma significantly lower than that in nasopharyngitis(χ~2=4.60, P=0.032).3. The positive rates of CD8~+: the positive rate in nasopharyngeal carcinoma is 77.77%, including 24cases of +(53.33%),9 cases ++(20.00%) and 2 cases of +++(4.44%); the positive rate in nasopharyngitis is 78.79%, including 22 cases of +(66.67%),4 cases of ++(12.12%) and 0 cases of +++(0%). There is no significant difference between these two groups(χ~2=3.51, P=0.319).4. The positive rates of CD20+: the positive rate in nasopharyngeal carcinoma is 48.88%, including 12cases of +(24.44%),6 cases ++(13.33%) and 5 cases of +++(11.11%); the positive rate in nasopharyngitis is 63.63%, including 10cases of +(30.30%), 7 cases of ++(21.21%) and 4 cases of +++(12.12%). There is no significant difference between these two groups(χ~2=1.89, P=0.595).5. The positive rates of COX-2: the positive rate in nasopharyngeal carcinoma is 86.67%, including 20 cases of +( 44.44%) ,12 cases ++( 26.67% ) and 7 cases of +++ ( 15.56% ) ; the positive rate in nasopharyngitis is 93.94%, including 6 cases of +(18.18%),11 cases of ++( 33.33% ) and 14 cases of +++ ( 42.42% ) .The positive rate in nasopharyngeal carcinoma significantly lower than that in nasopharyngitis(χ~2=10.31, P=0.016).6. Spearman correlation analysis was used to explore the correlation of COX-2 to CD3, CD4, CD8, and CD20 in nasopharyngeal carcinoma. The positive expression of COX-2 was correlated to that of CD4(R=0.464,P=0.023) ,but was not correlated to CD3, CD8 and CD20 ( R=0.245, P=0.249; R=-0.102, P=0.634; R=0.000, P=0.998)Conclusion:1. The number of CD3+ cell is more in nasopharyngeal carcinoma than in nasopharyngitis, with no significant difference about CD20~+ cells, which indicates that there is immunity dysfunction in nasopharyngeal carcinoma, especially cell immunity dysfunction.2. The number of CD3, CD4 in NPC is less than that of in CN, but that of CD8 between these two groups shows no difference. It makes clear that declining of the ratio of CD4~+/ CD8~+ maybe correlated with the local immunity dysfunction in pharynx nasalize.3. The positive rates of COX-2 expression are correlated to CD4, which indicates that CD4 can regulate the expression of COX-2 by some way.更多还原