【作者】 张丝媛；

【导师】 唐利立；

【作者基本信息】 中南大学 ， 外科学， 2012， 博士

【摘要】 目的抑癌基因p53与乳腺癌细胞生长、凋亡和耐药有密切联系。本实验将比较PCR-SSCP (聚合酶链反应-单链构象多态性)法与临床上常用的免疫组化法检测p53基因突变的结果是否一致,观察p53基因突变与乳腺癌常用临床病理指标的关系,并评估其与蒽环类及紫杉类为基础的新辅助化疗短期疗效的关系,初步探索其作为新辅助化疗疗效预测因子的可能性。方法本实验选取中南大学湘雅医院乳腺科2011年9月至2011年12月间初治的确诊为ⅡA、ⅡB及ⅢA期乳腺癌患者共65例为研究对象,随机行EC(表柔比星+环磷酰胺)、TC(多西他赛+环磷酰胺)或TAC(多西他赛+吡柔比星+环磷酰胺)方案新辅助化疗三周期,经外周血PCR-SSCP法和免疫组化法检测p53基因突变,并于第三周期化疗结束后使用触诊及B超评估治疗疗效。结果1.p53基因突变多见于乳腺癌临床分期ⅢA期、腋窝淋巴结转移阳性和ER表达阴性患者中(p值分别为0.047、0.032和0.036),而与患者年龄、绝经状况、肿瘤组织学分级、PgR、C-erbB2及Ki67表达无关。2.分子亚型中,p53基因突变在三阴性亚型乳腺癌中较多见(9%),基因未突变在luminal B亚型中较多见(34%)(p=0.032)。3. PCR-SSCP基因检测与免疫组化方法检测所得p53突变结果基本吻合(p<0.001),且一致性中等偏高(kappa=0.670)。4.p53基因突变患者对EC、TC及TAC方案新辅助化疗均不敏感(p=0.62),未突变患者对TAC方案化疗较其他两种敏感(p=0.025)。5.回归方程显示,乳腺癌新辅助化疗疗效与腋窝淋巴结转移、C-erbB2表达和p53基因突变有关,与年龄、月经状态、临床分期、PgR和Ki67表达无关(p<0.05)。其中ER和p53为保护因素(b<0)。结论1.p53基因突变多见于乳腺癌临床分期ⅢA期、腋窝淋巴结转移阳性和ER表达阴性患者中。分子亚型中,p53基因突变在三阴性亚型乳腺癌患者中较多见,而未突变在luminalB亚型患者中较多见。2.p53基因突变患者对EC、TC及TAC方案新辅助化疗不敏感,未突变患者则对TAC方案新辅助化疗有较高的敏感性,可能为指导实际临床用药提供一定的理论依据。3. PCR-SSCP方法与免疫组化法检测p53基因突变结果的一致性较高,且有一定的推广价值。更多还原

【Abstract】 ObjectiveTumor suppressor gene p53has been implicated in growth and apoptosis of breast tumor cells and resistance to chemotherapy. This study was performed to further access the differences between Immunohis-tochemistry(IHC), the most common way to detect p53gene mutation in clinical research, and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis in the inspection of p53mutation in breast cancer patients. In addition, we tried to figure out the relationship between the mutation and some clinical histologic indexes. Then, we had conducted the study of the prognostic value of p53mutation to chemotherapy response of anthracycline and taxanes, in order to evaluate the significance of the gene mutation for short term outcomes.MethodSixty-five patients with phase ⅡA, ⅡB and ⅢA breast cancer received neoadjuvant chemotherapy treatment of EC or TC or TAC scheme for three cycles. IHC and plasma PCR-SSCP were used to detect the mutation of p53gene while ultrasound was used to evaluate the effectiveness of neoadjuvant chemotherapy.Result1. Mutation of p53gene was directly relative to IIIA stage of TNM classification, metastasis of axillary nodes and negative of ER expression (p=0.047,0.032, and0.036), rather than age, menstrual condition, tumor histological grade, and expression of PgR, C-erbB2and Ki67.2. Among molecular subclasses of breast cancer, p53gene mutation was more common in thiple-negative subtype (9%) while non-mutation in luminal B subtype (34%).3. The result of PCR-SSCP was familiar with IHC in detecting p53gene mutation, and the similarity was high.4. None of the tumors with p53mutation were response to EC, TC or TAC scheme. In the contract, tumors without p53mutation showed a good response to TAC scheme (p=0.025).5. A significant correlation with neoadjuvant chemotherapy efficiency and axillary node metastasis, expression of ER, C-erbB2and p53mutation was found. Meantime, age, menstrual condition, TNM classification, and expression of PgR and Ki67were not correlation with the response to treatment (p<0.05). And ER and p53were regarded as protective factors of breast cancer (b<0).Conclusion1. Mutation of p53gene was directly relative to IIIA stage of TNM classification, metastasis of axillary nodes and negative of ER expression. Also, p53gene mutation was more common in thiple-negative subtype breast cancer, while non-mutation was in luminal B subtype.2. None of the tumors with p53mutation were response to EC or TC or TAC scheme. In the contract, tumors without p53mutation showed a good response to TAC scheme, which might provide a guide for future clinical therapy.3. PCR-SSCP analysis was just as the effective as IHC in detecting p53gene mutation.更多还原