MDR1、GSTP1基因遗传变异与上皮性卵巢癌患者铂类化疗临床预后关系的研究

发布时间：2018-01-19 20:38

  本文关键词： MDR1 GSTP1 单核苷酸多态性（SNP） 卵巢上皮性癌 铂类化疗 临床预后　出处：《河北医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：卵巢恶性肿瘤是女性生殖系统常见的三大恶性肿瘤之一，其中最常见的妇科恶性肿瘤为上皮性卵巢癌（epithelial ovarian cancer，EOC)，它在导致女性死亡的病因中排名第七位，患者5年生存率约30%[1]。EOC患者的治疗主要包括手术和术后的辅助化疗。以铂类为基础的联合化疗是EOC患者术后的一线治疗方案，但患者对铂类的耐药严重影响了化疗疗效，成为治疗EOC的一大障碍。铂类耐药可能受累于多种机制，个体遗传因素所导致的对药物的反应差异在治疗中起着重要作用。 P-糖蛋白是一种膜转运蛋白，它位于细胞浆膜，可将细胞内的化疗药物泵出细胞外，使细胞内药物浓度下降，增加了肿瘤细胞的耐药性。P-糖蛋白表达水平的高低可能导致上皮性卵巢癌患者对铂类药物的应答和预后的差异，而编码P-糖蛋白的基因即为多药耐药基因1（multidrugresistance1，MDR1）。同样，细胞对药物代谢解毒功能的改变亦与铂类耐药密切相关。铂类药物进入细胞后在谷胱甘肽-S-转移酶P1（glutathione-S-transferases P1, GSTP1）的催化下可与谷胱甘肽（glutathione,GSH）形成复合物，该复合物毒性降低且易于被转运出细胞，有利于铂类药物从尿液或胆汁中排出，从而达到代谢解毒的功能[2]。位于MDR1、GSTP1基因的遗传变异可能通过改变其所编码蛋白质的表达，进而影响患者对铂类药物的应答及临床预后。 本研究拟探讨MDR1和GSTP1基因遗传变异与上皮性卵巢癌患者铂类耐药及临床预后的关系。旨在寻找上皮性卵巢癌患者铂类化疗的分子标志物，为临床的“个体化”治疗提供重要的实验数据。 方法：采用回顾性病例研究方法，病例为2001年12月至2010年6月期间在河北医科大学第四医院妇科进行治疗的上皮性卵巢癌患者（225例）。术前采集外周静脉血5ml，以蛋白酶K消化-饱和氯化钠盐析法提取基因组DNA，采用连接酶检测反应（LDR）检测上皮性卵巢癌患者的MDR1基因C3435T、G2677T/A和GSTP1基因A313G这3个SNP位点的基因型和等位基因频率分布情况。所有患者均经过3年随访。判定患者临床预后的指标包括患者对铂类化疗的应答、总生存时间（overallsurvival，OS）和疾病无进展生存期（progression-free survival，PFS）。 数据分析采用SPSS13.0统计软件进行处理。采用χ2检验检测基因型和等位基因与铂类化疗应答的关系；以logistic regression方法计算相对风险度的比值比(odds ratio, OR)及其95%可信区间(confidence interval, CI)；生存分析采用Kaplan-Meier法，各因素水平间比较用Log-rank及Breslow分析，Cox回归模型进一步分析预后相关因素（患者年龄、肿瘤分化程度、病理类型、临床分期、残余病灶）。P＜0.05作为差异有显著性的标准。 结果： 1MDR1基因C3435T、G2677T/A和GSTP1基因A313G三个多态位点的基因型和等位基因频率分布在铂类化疗应答组与不应答组之间均无统计学差异（P＞0.05）。 2MDR1基因C3435T与上皮性卵巢癌患者临床生存预后的关系：患者以复发组和未复发组分类进行生存分析，C/C、C/T、T/T三种基因型在复发组和未复发组的频率分布分别为35.8%、47.0%、17.2%和26.4%、57.1%、16.5%，经Kaplan-Meier单因素分析两组比较无统计学差异（P0.05），，进一步的Cox回归多因素分析亦无统计学差异（P0.05）；以生存组和死亡组分类，三种基因型频率分布分别为31.6%、52.9%、15.4%和32.6%、48.3%、19.1%， Kaplan-Meier单因素分析和Cox回归多因素分析显示均无统计学差异（P0.05）。 3MDR1基因G2677T/A与上皮性卵巢癌患者临床生存预后的关系：G/G、G/T、A/G、A/A、A/T、T/T六种基因型在复发组和未复发组的频率分布分别为：17.9%、29.1%、14.2%、2.2%、18.7%、17.9%和8.8%、35.2%、13.2%、2.2%、17.6%、23.1%。以T/T基因型和非T/T基因型进一步分类，在复发组和未复发组的频率分布分别为：17.9%、82.1%和23.1%、76.9%。Cox回归多因素分析，复发组和未复发组比较具有统计学意义（P=0.01，HR=3.27，95%CI=1.36-7.88）。以生存组和死亡组分类，频率分布分别为：14.0%、33.1%、14.0%、1.5%、17.6%、19.9%和14.6%、29.2%、13.5%、3.4%、19.1%、20.2%，经Kaplan-Meier单因素分析和Cox回归多因素分析，6种基因型的频率分布无统计学差异。 4GSTP1基因A313G与上皮性卵巢癌患者临床生存预后的关系：患者以复发组和未复发组分类进行生存分析， A/A、A/G、G/G三种基因型的频率分布分别为：56.7%、38.1%、5.2%和64.8%、31.9%、3.3%，以Kaplan-Meier单因素分析和Cox回归多因素分析均无统计学差异（P0.05）；三种基因型在生存组和死亡组的频率分布分别为60.3%、37.5%、2.2%和59.6%、32.6%、7.9%，以Kaplan-Meier单因素分析和Cox回归多因素分析比较亦无统计学差异（P0.05）。 结论： 1MDR1基因C3435T、G2677T/A和GSTP1基因A313G多态可能与上皮性卵巢癌患者铂类化疗应答无关。 2MDR1基因C3435T多态可能与上皮性卵巢癌患者铂类化疗后的复发和生存时间无关。 3MDR1基因G2677T/A多态性可能与中国北方妇女上皮性卵巢癌患者铂类化疗后的复发相关，但与其生存时间无关。 4GSTP1基因A313G多态可能与上皮性卵巢癌铂类化疗患者的复发和生存时间无关。
[Abstract]:Objective: ovarian cancer is one of the female reproductive system of three malignant tumor, one of the most common gynecological malignant tumor of epithelial ovarian cancer (epithelial ovarian, cancer, EOC), which causes death in women ranked seventh, 5 year survival rate of patients with treatment about 30%[1].EOC patients include surgery adjuvant chemotherapy and after the operation. Platinum based chemotherapy is the first-line treatment for EOC patients, but patients resistant to platinum has seriously affected the efficacy of chemotherapy, become a major obstacle in the treatment of EOC. Platinum resistance may be tired from a variety of mechanisms, the reaction to the drug because of the difference of individual genetic factors it plays an important role in the treatment.
P- glycoprotein is a transmembrane protein, which is located in the cell membrane, can be used for cell drug efflux pump in cells, the intracellular drug concentration decreased, increased tumor cell drug resistance.P- glycoprotein expression level may lead to differences of platinum based chemotherapy response and prognosis of patients with epithelial ovarian cancer, and encoding P- glycoprotein gene is multidrug resistance gene 1 (multidrugresistance1, MDR1). Also, the change of cell on drug detoxification function and platinum resistance are closely related. The platinum drugs enter cells in glutathione -S- transferase P1 (glutathione-S-transferases P1 GSTP1) catalyzed with glutathione (glutathione. GSH) to form complexes, the complexes reduced toxicity and easy to be transported out of the cell, to platinum drugs excreted in urine or bile, so as to achieve the function of [2]. detoxification is located in MDR1, G The genetic variation of the STP1 gene may be altered by changing the expression of the encoded protein, thereby affecting the patient's response to platinum drugs and the clinical prognosis.
The aim of this study is to explore the relationship between MDR1 and GSTP1 genetic variation and platinum resistance and clinical prognosis in patients with epithelial ovarian cancer. It is to find molecular markers of platinum based chemotherapy in epithelial ovarian cancer, and provide important experimental data for clinical individualized treatment.
Methods: a retrospective case study method, case for the period from December 2001 to June 2010 in patients with epithelial ovarian cancer treated in the fourth hospital of Hebei Medical University Department of gynaecology (225 cases). Preoperative collection of peripheral venous blood 5ml by proteinase K digestion saturated sodium chloride salting out method to extract genomic DNA, using the ligase detection reaction (LDR) detection the MDR1 gene of C3435T in patients with epithelial ovarian cancer, G2677T/A gene and GSTP1 gene A313G 3 SNP loci genotype and allele frequency distribution. All patients were followed over 3 years. To determine the prognosis of the patients were on platinum based chemotherapy response, overall survival time (overallsurvival, OS) and disease progression free survival (progression-free, survival, PFS).
Data were analyzed by SPSS13.0 software. The relationship between 2 test was used in detection of genotype and allele and response to platinum based chemotherapy; logistic regression method to calculate the ratio of the relative risk ratio (odds ratio, OR) and 95% confidence interval (confidence, interval, CI); survival analysis using the Kaplan-Meier method, Log-rank analysis and Breslow comparison between the level of various factors, Cox regression model to analyze the prognostic factors (age, tumor differentiation, pathological type, clinical stage, residual lesions).P < 0.05 as there was a significant difference between the standard.
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