乙醛脱氢酶1在乳腺癌中的表达及临床意义.docx

上海交通大学硕士学位论文乙醛脱氢酶1在乳腺癌中的表达及临床意义姓名：周凌申请学位级别：硕士专业：外科学（普通外科）指导教师：李克20090501SCCHNHSP9017-DMAGsiRNADEAB头颈部鳞状上皮细胞癌热休克蛋白90热休克蛋白90功能特异性抑制剂小干扰RNA二氨基苯卬醛squamous cell carcinoma of the head and neck heat shock protein 9017-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochloride small interfering RNAdiethylaminobenzaldehyde乙醛脱氢酶1在乳腺癌中的表达及临床意义摘要目的 探讨干细胞标记物:乙醛脱氢酶1 （aldehyde dehydrogenas1,ALDH1）在乳腺癌中的表达与临床意义方法 收集上海交通大学附属第一人民医院2003年1月至2008年3月乳腺癌病例92例采用免疫组织化学染色法检测所有病例乳腺癌组织中 ALDH1的表达，并结合临床病理参数进行无病生存期分析所有实验数据 用%2检验或Fisher精确检验，生存分析采用Kaplan-Meier法，采用 log-rank检验比较生存曲线的差异。

采fflCox比例风险模型分析影响预后 的因素采用SPSS 13. 0统计软件包进行统计学处理，以PO.05为差异有 统计学意义结果 ALDH1的阳性表达率为18.5% o ALDH1在乳腺癌组织中的阳性表达与PR、c-erbB-2有关（P<0.05）,与患者年龄、肿瘤大小、临床分期、 是否有淋巴结转移无关（P>0.05）ALDH邛印性组患者2年无病生存率显 著低于阴性组（PV0.05）,接受CEF或TE方案化疗和内分泌治疗的ALDH1 阳性组患者2年无病生存率显著低于阴性组（PV0.05）,转移病例中， ALDH1阳性组2年无病生存率显著低于阴性组（PV0.05）结论 ALDH1表达阳性的乳腺癌细胞具有乳腺癌干细胞的部分特性：ALDH1阳性细胞占肿瘤细胞极少部分（<10%）；子代（转移灶）与母代肿 瘤的病理特征相似；ALDH1表达阳性患者对化疗，内分泌治疗等常规辅助 治疗耐药，提示ALDH1阳性细胞是-•种“类干细胞”或具有干细胞潜能的细胞群落乳腺癌ALDH1表达阳性患者2年无病生存率显著低于乳腺癌ALDH1表达阴性患者，证实乳腺癌ALDH1表达阳性患者预后较差使用ALDH1免疫组化方法来鉴别乳腺癌组织内“类干细胞”群落，促进了肿瘤 干细胞概念在临床的应用。

同时为今后乳腺癌靶向治疗提供了新的方向关键词：乳腺肿瘤，干细胞，乙醛脱氢酶1,预后，免疫组织化学EXPRESSIONS OF ALDEHYDE DEHYDROGENAS 1 INBREAST CANCER AND THEIR CLINICAL SIGNIFICANCEABSTRACTObjective The purpose of this study is to investigate the expression and clinical significance of aldehyde dehydrogenas 1 (ALDH1) in breast cancer・Methods The expressions of ALDH1 protein was examined by immunohistochemical staining in 92 breast cancer tissues・ The disease-free survival analysis of patients was evaluated based on the clinical follow-up data. The personal chi-square test was employed to examine the correlation between each variable and to investigate the statistically significant differences between molecular makers and clinicopathological parameters. The disease- free survival (DFS) was estimated based on the Kaplan-Meier survival analysis, and the association between variables and survival was analyzed statistically by the log-rank test. For multivariate analysis, the Cox regression model was used with 95% confidence intervals. All statistical analyses were carried out with SPSS(Statistical Package for the Social Sciences) for Windows version 13.0 software. The results were considered to be statistically significant at P<0.05.Results Of all these 92 cases, 17 cases (18.5%) were observed as positive for ALDH1 expression in primary tumors. Expressions of ALDH1 protein had significant correlation with PR, c-erbB-2(P<0.05) 9but no significant correlation were found between the expression of ALDH1 protein and age,tumor size.TNM stagejymph node status (P>0.05) .Patients with A1DH1 positive tumors had lower 2・year disease-free survival rate than that of those with ALDH1 negative tumors(p<0.05). Patients, who underwent chemotherapy and hormone therapy, with A1DH1 positive tumors had lower 2-year disease-free survival rate than that of those with ALDH1 negative tumors(p<0.05). In the cases with lymph node metastasis, the 2-year disease-free survival rate of patients with ALDH1 positive tumors was also lower than that of patients with ALDH1 negative (p < 0.05).Conclusion ALDH1 positive cells form tumors from very few cells, recapitulate the heterogeneity of the parental tumor, and are selectively resistant to chemotherapy and hormone therapy, all of which are hallmarks of cancer stem cells・ ALDH1 positive cells may be a small population of breast cancer stem-like cells ・ Patients with A1DH1 positive tumors had lower 2-year disease-free survival rate than that of those with ALDH1 negative tumors, and ALDH1 may be as a prognostic marker. ALDH1 immunostaining is a simple method for identifying similar breast cancer stem cell-like populations・ This marker may facilitate the application of cancer stem cell biology to clinical practice. The development of ALDH1 as a potential target for vaccination therapy will offer new hope to women with breast cancer・KEY WORDS： breast neoplasms, stem cells, aldehyde dehydrogenas 1, prognosis, immunohistochemistry乙醛脱氢酶1在乳腺癌中的表达及临床意义亠八 —冃IJ 51997年由Dick团队发现人类白血病起源于极小部分白血病干细胞，并且这种细 胞亚群能在NOD/SCID (nonobese diabetic /severe combined immunodeficient, 非肥胖糖尿病/重症联合免疫缺陷)小鼠中形成白血病，首次用实验证据证实肿瘤干 细胞的假设［1］。

2001年Taipale J提出“肿瘤十细胞”理论［2］2003年A1-Hajj［3］进一 步拓展了这个概念，他的团队发现人类乳腺癌包含了一小部分表面分子标记为 CD44+/CD24-/lin-的细胞群拥有十•细胞特性，第一次证实了乳腺癌干细胞的存在随 后肿瘤干细胞的理论一直被不断的论证和修定如今大量种类的的肿瘤干细胞被鉴 定及分离出来，包插脑癌，前列腺癌，恶性黑色素瘤，结肠癌，胰腺癌，头颈部鳞 癌等［4-1 l］o 一些研究支持这样一种假设，即肿瘤干细胞来源于正常的细胞起源(干细 胞或起始细胞)，由于致癌因素导致了正常细胞的恶性转化［12, 13］于是人们开始志 力于寻找一种建立于细胞功能的肿瘤干细胞的标记物，这种标记物在起源细胞内存 在，并能遗传到同样起源的致变的恶性干细胞内乙醛脱氢酶1 (aldehyde dehydrogenas 1,ALDH1)就是这样一个候选指标乙醛脱氢酶(aldehyde dehydrogenas ,ALDH) ［14］是•组能将乙醛氧化成乙酸的 同工酶，能保护细胞避免醛过氧化物的损害John P Chute］ 15］等研究证明了ALDH1在 干细胞的早期分化中起了重要的作用。

有报道显示ALDH高度聚焦在人类造血干细胞 (hematopoietic stem cells , HSCs) ［16］和神经系统十•细胞［4, 5］,在多发性骨髓瘤 和急性粒细胞白血病中也发现肿瘤细胞的ALDH活性有所增高［17, 18］o于是ALDH活性成 为了正常和恶性肿瘤干细胞的共同。