【作者】 张军；

【导师】 曹月敏；

【作者基本信息】 河北医科大学 ， 外科学， 2003， 硕士

【摘要】 目的：乳腺癌是最常见的恶性肿瘤之一, 在医疗科技日益发展的今天其发病率和死亡率仍较高。 1894年Halsted创立了经典的乳腺癌根治术，随后又出现了改良根治术，近年来更着力发展了化学药物治疗、放射治疗、免疫治疗和生物学治疗等新方法，但疗效始终不甚理想。其原因主要是因为绝大多数患者就诊时已属临床中、晚期乳腺癌，已经失去了手术根治的最佳时机，即使采用现代的综合治疗措施其生存率和生活质量也不能让人满意。因此，早期诊断，即尽可能在肿瘤局限于乳腺局部尚未发生转移之前就做作出明确诊断便成了治疗的关键。鉴于目前常用的检查方法如乳腺B超、X线、热像图及针吸活检等对早期诊断效果不佳，人们开始探索新的乳腺癌的诊断途径，希望对临床有所帮助。选择素是细胞粘附分子的一大类成员，包括P-选择素、E-选择素和L-选择素，在体内的主要功能是在炎症反应中介导白细胞血小板与内皮细胞粘附，从而促进白细胞向炎症区域集中。近年来的研究发现，选择素及其受体S-lewisx，S-lewisα，PSGLE-1等在某些恶性肿瘤细胞表面也有分布，因此推测选择素的异常表达可能与恶性肿瘤的存在有一定的关系。为探讨乳腺癌的发生发展和转移与可溶性P-和E-选择素的关系，我们设计了本研究：检测血浆可溶性P-和E-选择素在正常<WP=4>人、乳腺癌患者（包括已发生腋窝淋巴结转移和未发生腋窝淋巴结转移的两组病人）中的含量并对比其差异，借以阐明二者的联系，希望能为乳腺癌的早期诊断和病程分期提供新的有益的临床参考指标。方法：1 病例选择：乳腺癌患者52例，均为女性，来自河北省人民医院腺体外科2001年6月至2003年1月的住院病人，年龄42-65岁，平均（48±9.7）岁，患者均经术后病理检查证实，病理类型包括腺癌、导管内癌、乳头状癌、鳞癌、浸润性小叶癌和髓样癌，其中腋窝淋巴结病理检查证实已发生转移的32例，未转移的20例。患者均无心脑血管疾病，各重要脏器功能正常。同时设置正常对照组30例，均为女性，年龄22-53岁，平均（45±8.6）岁，来自河北省人民医院健康体检人员，无心脑血管疾病，各重要脏器功能在正常范围，无急、慢性感染性疾病，其它化验指标符合健康标准。2标本采集：乳腺癌患者及正常对照组均清晨空腹采静脉血3ml，EDTA抗凝，3000转/分离心10分钟，分离血浆后置-70℃冰箱储存。3检测方法：可溶性P-和E-选择素用酶联免疫吸附试验（ELISA法）检测，所需试剂由北京晶美公司提供，由实验人员严格按操作程序检测，所需设备由河北省人民医院临床科研中心提供。4统计学方法：计量资料数据以均数±标准差（(x ( s）表示，各组数据均进行正态性检验，组间进行方差齐性检验，组间比较用方差分析法，以P<0.05作为有显著性差异，所有数据均由SPSS10.0统计学软件处理。结果： 1乳腺癌患者血浆可溶性P-选择素水平（301.2<WP=5>±113.8ng/ml）较正常人（93.13±35.05ng/ml）明显升高，差异非常显著（P<0.01），已发生腋窝淋巴结转移的患者其表达水平（331.6±110.9ng/ml）较未发生转移的患者（249.5±98.32ng/ml）升高且有统计学意义（P<0.05）。2乳腺癌患者血浆可溶性E-选择素水平（32.53±12.66ng/ml）较正常人（14.68±5.12ng/ml）明显升高且有统计学意义（P<0.01），已发生腋窝淋巴结转移的患者可溶性E-选择素表达水平（37.11±12.41ng/ml）较未发生转移者的表达水平（25.18±9.23ng/ml）升高，结果有统计学差异（P<0.05）。结论： 通过对乳腺癌患者血浆可溶性P-和E-选择素的含量与正常人的含量的对比研究，以及对已发生腋窝淋巴结转移患者和未发生转移患者的表达量的比较，可得出以下结论: 1.乳腺癌患者因大量恶性肿瘤细胞的异常代谢产物以及机体对肿瘤的免疫学反应激活大量的内皮细胞和血小板，致使膜型P-和E-选择素表达增加，此时外周血用ELISA法可测出可溶性P-和E-选择素的明显改变。2.已发生腋窝淋巴结转移的乳腺癌患者体内可溶性P-和E-选择素表达量较未发生转移的患者明显升高，说明P－和E－选择素在乳腺癌的腋窝淋巴结转移中起着重要作用，二者具有明显的相关性。3.可溶性P-和E-选择素具有检测容易、灵敏度高、成本低等特点，如果对其进行大样本的深入研究，可望成为早期诊断乳腺癌和判断病程分期的一个有益的参考指标。更多还原

【Abstract】 Objective: Breast cancer is a disease characterized by unrestrained and excessive multiplication of breast cell. It is one of the most conventional malignant tumours in human body. Surgeons will often in face of the diagnosis and therapy of this disease. Today. with the developed technology of medical, surgeons still think that they can not succeed in healing breast cancer. In 1894, the American surgeon, Halsted set up the classical extirpative surgery of breast cancer to cure this disease, after that, people develop all kinds of extirpative or palliative surgery to cure the early and late stage patients, In recent years, doctors also use chemotherapy and radiation therapy to treat this cancer, but the result is not satisfactory, because most breast cancer patients can not find out that she or he has already got this deadly disease in early stage. When they fell not very well and go to hospital, they have already lost the best chance to save themselves. So early diagnose of breast cancer is very important. But today’s method of diagnosis (including X-ray scan, excisional biopsy , incisional biopsy, exfoliative cytology, et al)can not diagnose the early stage breast cancer. In this condition. Doctors have to seek for<WP=8>new method of diagnosis. So they think of selectins. Selectin is a family of the cell adhesion molecule, including platelet-selectin, endothelium-selectin and leukocyte-selectin. Ihe function is to introduce the adhesion of platelet and leukocyte with the endotheliocyte. In recent years, doctors learned that selectins and their ligands: S-Lewisx S-Lewisа and PLGSE-1 also can be found in the surface of some kinds of malignant tumour cells .they think that maybe selectin will have some kinds of relationship with malignant tumour. In order to study the relationship between soluble P-selectin、 soluble E-selectin and breast cancer， we design this experiment: |determing the concentration of soluble P-selectin and soluble E-selectin in breast cancer and healthy people. trying to find out the relationship between them and hope to find a new method of diagnosIs in early stage breast cancer.Mothod : 1 A total of 52 patients with breast cancer were recruited in this study. They were all come from He Bei Province Hospital. The ages ranged from 42 to 65. They were further divided into late stage cancer group (32patients) and early stage cancer group (20 patients). At the same time 30 healthy examinations were chosed . the ages ranged from 32 to 57，also come from He Bei Province Hospital . 2 Sample collection: 3 ml elbow vein blood were taken from both the breast cancer and healthy people. It was soon put into anticoagulative test tube， rotating with 3000r/m for 10 minutes， separate plasma and stored into-70℃freezer. 3<WP=9>Soluble P-selectin and soluble E-selectin were determined by enlyme-linked immunosorbent assay, the reagent needed were came from Jing Mei Company (Bei jing). The assay was carried in the Clinical Scientific Research Center of He Bei Province Hospital. 4 All the data were showed in (x ( s and the calculation of this test was carried by computer with SPSS10.0 statistical programme.Result: 1 The concentration of soluble P-selectin in breast cancer(301.2±113.8ng/ml) is higher than that in healthy people(93.13±35.05ng/ml), the soluble P-selectm level in late stage cancer(331.6±110.89ng/ml)is higher than the level in early stage breast cancer(249.5±98.32ng/ml) 2 the concentration of soluble E-selectin in breast cancer(32.53±12.66ng/ml)is higher than that in healthy people (14.68±5.12ng/ml), the soluble E-selectin level in late stage cancer (37.11±12.41ng/ml)is higher than the level in early stage breast cancer(25.18±9.23ng/ml).Conclusion: According to the study of the soluble P-and E-selectin in breast cancer, it has been reached the conclusion as follows 1.There are many malignant tumour cells in breast cancer patient. The metabolism of malignant tumour cells activate so many leukocytes and platelets, So the soluble P-selectin and E-select更多还原