【作者】 鲍慧铮；

【导师】 王丽；

【作者基本信息】 东北师范大学 ， 细胞生物学， 2008， 博士

【摘要】 乳腺癌是女性中最常见的恶性肿瘤之一,全球每年发病人数超过100万。资料证明,全世界乳腺癌发生率正以每年0.2%-3%的幅度上升,1992年美国乳癌发病达180000例,发病率达111.4/10万人。据统计,我国1994年乳腺癌新增病例110635例,发病率20.2/10万人,死亡27343人。目前每年因乳腺癌失去乳房或夺去生命的绝对数字都在不断上升。随着人们防癌知识的普及与提高,医学科学的进步、医疗设备的改善,乳腺的多种疾病,尤其是乳腺癌的防治工作已经取得了显著的成效。乳腺癌的预防有了新的苗头,早诊技术推陈出新,乳腺癌综合治疗效果已经有所提高。自20世纪70-80年代,大量的临床证明化疗能明显提高乳腺癌病人的生存率、改善病人的生存质量,化学药物治疗(简称化疗)是一种必要的全身性辅助治疗,它与手术治疗、放射治疗及内分泌(激素)治疗乳腺癌一样,都是不可缺少的,且具有同等重要的临床价值。化疗已成为浸润性乳腺癌的主要疗法之一。新辅助化疗目前被临床广泛应用于晚期乳腺癌的系统治疗。应用新辅助化疗,能够显著降低肿瘤分期,提高保乳手术几率。尽管乳腺癌的新辅助化疗已经在国内外临床上施行很长时间,然而对于新辅助化疗过程中一些肿瘤标志性分子的研究还有待深入,国内外学者对一些标志性分子的功能及表达规律还存在很大争议。p53抗体在肿瘤早期以及一些癌前病变即能检出,大量资料表明,p53抗体的出现在恶性肿瘤的发生过程中是一个非常早期的事件。在本实验中,我们利用基因重组技术,成功构建了带有6His纯化标签的人野生型全长p53蛋白的原核表达质粒;对工程菌表达蛋白的条件进行优化后,对p53蛋白进行了大量表达,经镍离子金属螯合亲和层析纯化处理,得到了全长p53蛋白的纯化产物;我们利用ELISA技术,对86名乳腺癌患者血清进行了检测,全长p53蛋白检测体系的特异度分别为95%。在本实验中,我们利用ELISA技术,分别以重组p53蛋白为固相包被抗原,棋盘滴定法确定了最佳包被浓度和最佳血清稀释倍数后,对400名健康人血清进行了检测,确定了各自体系的cut-off值;又对86名乳腺癌患者血清进行了检测,全长p53蛋白检测体系的特异度分别为95%。我们比较了血清p53抗体表达与临床用药之间的相关性,确定p53抗体与某些临床参数具有显著相关性。通过研究发现,血清p53抗体具有一定的预测价值,化疗前p53抗体阴性的患者对葸环类药物敏感,肿瘤应答率高,治疗效果明显。而化疗前血清p53抗体呈阳性的患者,对蒽环类药物具有抗性,治疗效果不理想。以上实验结果充分表明p53抗体在肿瘤预后方面具有显著意义。表明p53抗体在肿瘤预后方面具有显著意义。因此,开展p53抗体检测,探索肿瘤患者血清p53抗体表达与患者临床资料、肿瘤治疗方案选择以及其他肿瘤标志物表达的相关性研究,是一项非常有利于乳腺癌新辅助化疗临床治疗、用药选择和预后判断的工作。更多还原

【Abstract】 Breast carcinoma is one of the most common malignancies in women. Every year, more than 1,000,000 people are sufferring from this disease worldwide. Previous data indicated that the incidence of breast cancer was increasing by 0.2% to 3% every year. In 1992, there were 180,000 breast carcinoma patients in the US, and the incidence was as high as 1.11%. According to statistical data, in 1994, 110,635 new entrants suffered from breast carcinoma, and the incidence was 0.2%. Moreover, 27,343 people dead from this disease. The number of patients who lost breast and dead continues increasing year by year. Following popularization and enhancement knowledge of cancer prevention, improvement of medical science, amelioration of medical installation, several mastopathia could be controlled, especially breast carcinoma. Recently, as more and more technical innovations are used to prevention and earlier diagnosis, combined therapeutic efficacy on breast carcinoma has made significant achievement.In the 70^th and 80^th of 20 century, much clinical evidence indicted that chemotherapy could promote higher survival rate and quality. As a pantosomatous adjunctive therapeutic strategy, chemotherapy is as important as surgery, radiotherapy and endocrine therapy to treat breast cancer, they have equally clinical value. Chemotherapy has became one principal method to therapy invasive breast cancer. Neoadjuvant chemotherapy（NAC） is commonly used as a systemic treatment of locally advanced breast cancer. This treatment is usually given to downstage tumors and promotes higher conservative breast surgery rates. Although neoadjuvant chemotherapy for breast cancer has been used in clinic for a long time, it’s unclear that the function of tumor biomakers during the treatment process of NAC.Many investigations have suggested that the anti-p53 antibodies（Abs） would be detected in the earlier stage of neoplasia. In this experiment, used the gene recombination technology, the wild-type p53 protein was expressed and purified. Using the ELISA approach, we detected 86 patients with breast cancer, and the respectively was 95%. The relationship between the expression of serum p53 antibody and patients’ characteristics was investigated, and we found some correlation between the p53 Abs expression and these characteristics. Besides, the result showed that, before the NAC treatment, patients who were p53 Abs negative would obtain a better response to the anti-cancer drugs of anthracycline, while the positive would obtain a bad response. Therefore, the detection of anti-p53 Abs, studying of the relationships between anti-p53 Abs and patients’ characteristics, treatment schemes and other tumor biomakers will do a lot of good to the breast cancer’s clinical diagnosis, prediction and anti-cancer drug’s choice.更多还原