卵巢癌化疗敏感性相关生物标记物的研究

发布时间：2018-02-17 00:17

本文关键词： 卵巢癌生物标记物耐药无进展生存期总生存期　出处：《华中科技大学》2014年博士论文　论文类型：学位论文

【摘要】：目的探索与上皮性卵巢癌化疗敏感性相关的生物标记物,并研究其与卵巢癌预后之间的相关性,找出能够预测化疗敏感性及预后的因子,为卵巢癌个体化治疗提供更多的证据。方法 1.通过生物信息学方法选取20个卵巢癌化疗耐药相关的生物标记物,其中包括药物代谢/转运相关：MDR1, CTR1, HIF-1α, MRP, PXR; DNA损伤修复相关：PARP1, ERCC1, BRCA1, BRCA2;细胞存活/凋亡通路相关：AKT1, EZH2, EGFR, VEGF, BCL-2, HDAC1, SURVIVIN, TNF-a, HSP27, RGS17, RASSF1A。 2.选取武汉协和医院2008—2013年手术病理证实为上皮性卵巢癌的病人42例,另有5例正常卵巢组织作为对照。病人术前未行放化疗,术后接受TC方案化疗(紫杉醇+卡铂,静脉化疗)。根据化疗反应性分为耐药组和敏感组。组织芯片技术免疫组化染色检测各目的因子在卵巢癌患者组织中的表达情况。 3.激光显微切割技术获取纯化的卵巢癌组织,qRT-PCR技术检测目的因子在mRNA水平的表达情况。结果 1.免疫组化结果显示,选取的20个标记物中,有14个因子(MDR1,AKT1,EZH2,PARP1, CTRL HIF1α, HDAC1, EGFR, VEGF, HSP27, ERCC1,BRCA1, BRCA2, PXR)的蛋白表达水平在耐药组和敏感组中有统计学差异。其中,CTR1在耐药组中低表达,敏感组中高表达；其余各因子在耐药组中均为高表达,敏感组中低表达。 2.ROC曲线分析显示,用于预测化疗敏感性时,上述14个因子的曲线下面积均达到0.7以上,其中PARP1和HDAC1的曲线下面积分别为0.904和0.894。 3.生存分析结果显示,MDR1,EZH2, PARP1,HDAC1,EGFR与患者PFS有关,蛋白表达水平越高,PFS越短；HDAC1, BRCAl与患者OS有关,蛋白水平越高,OS越短。 4.多因素分析结果提示,EZH2, PARP1,HDAC1,EGFR高表达是PFS的独立危险因素,HDAC1,BRCA1高表达是OS的独立危险因素。 5. qRT-PCR结果显示,MDR1,EZH2, PARP1,HDAC1的mRNA水平,耐药组均高于敏感组。利用ROC曲线分析其预测化疗反应性的效能,其曲线下面积均大于0.7,最高的HDAC1为0.932。 6.基于mRNA水平的生存分析结果显示,与免疫组化结果一致,MDR1,EZH2, PARP1,HDAC1,EGFR的mRNA水平同样与患者PFS相关,表达水平越高,PFS越短。多因素分析也提示,MDR1,EZH2, PARP1,HDAC1,EGFR是患者PFS的危险因素。结论 PARP1和HDAC1有希望作为预测卵巢癌化疗敏感性的生物标记物。此外,MDR1、EZH2、PARP1、HDAC1、EGFR是PFS的独立危险因子；HDAC1、BRCA1是OS的独立危险因子。
[Abstract]:Purpose. To explore the biomarkers related to the chemosensitivity of epithelial ovarian cancer and its correlation with the prognosis of ovarian cancer, to find out the factors that can predict the chemosensitivity and prognosis of ovarian cancer, and to provide more evidence for individualized treatment of ovarian cancer. Method. 1. Twenty biomarkers associated with chemoresistance of ovarian cancer were selected by bioinformatics. These include drug metabolism / transport correlation: MDR1, CTR1, HIF-1 伪, MRP, PXR; DNA damage repair correlation: PARP1, ERCC1, BRCA1, BRCA2; cell survival / apoptosis pathway associated with: AKT1, EZH2, EGFR, VEGF, BCL-2, HDAC1, SURVIVIN, TNF-a, HSP27, RGS17, RASSF1A. 2. Forty-two patients with epithelial ovarian cancer confirmed by operation and pathology from 2008 to 2013, and 5 normal ovarian tissues as control group, were selected from Wuhan Union Hospital in 2008.The patients did not undergo radiotherapy and chemotherapy before operation and received chemotherapy with TC regimen after operation (paclitaxel carboplatin), and the patients received paclitaxel carboplatin (paclitaxel carboplatin). According to the chemoreactivity, the patients were divided into two groups: drug resistant group and sensitive group. The expression of various target factors in ovarian cancer tissues was detected by immunohistochemical staining with tissue microarray technique. 3.The expression of target factor at mRNA level was detected by qRT-PCR in purified ovarian cancer tissue obtained by laser microdissection. Results. 1. Immunohistochemical results showed that the protein expression levels of MDR1AK T1AK-EZH2P1, CTRL HIF1 伪, HDAC1, EGFR, VEGF1, HSP27, ERCC1, BRCA1, BRCA2, PXR1 were significantly different between the resistant group and the sensitive group. The expression of other factors was high in the drug resistant group and low in the sensitive group. 2. ROC curve analysis showed that the area under the curve of the above 14 factors was more than 0.7 when used to predict chemosensitivity, and the areas under the curve of PARP1 and HDAC1 were 0.904 and 0.894, respectively. 3. Survival analysis showed that MDR1EZH2, PARP1HDAC1EGFR were related to PFS, the higher the protein expression level was, the shorter the HDAC1protein expression level was. The higher the protein level was, the shorter the HDAC1protein level was. The higher the protein level was, the shorter the HDAC1protein level was. 4. The results of multivariate analysis suggested that the high expression of EZH2, PARP1HDAC1EGFR was the independent risk factor of PFS. The high expression of HDAC1BRCA1 was an independent risk factor of OS. 5. The qRT-PCR results showed that the mRNA levels of EZH2, PARP1HDAC1 in MDR1 and PARP1HDAC1 were higher in the resistant group than in the sensitive group. The area under the curve was greater than 0.7 and the highest HDAC1 was 0.932 by ROC curve analysis. 6. The survival analysis based on mRNA level showed that the mRNA level of MDR1 / EZH2 and PARP1HDAC1HDAC1EGFR was also correlated with PFS, and the higher the expression level was, the shorter the mRNA level was. Multivariate analysis also suggested that MDR1HDAC1EGFR was the risk factor of PFS in patients with EZH2.The results of multivariate analysis also indicated that MDR1HDAC1EGFR was the risk factor of PFS. Conclusion. PARP1 and HDAC1 are promising biomarkers for predicting chemosensitivity of ovarian cancer. In addition, MDR1EZH2PARP1HDAC1EGFR is an independent risk factor for PFS. HDAC1BRCA1 is an independent risk factor for OS.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.31

【共引文献】