本文關(guān)鍵詞： MDR1 GSTP1 單核苷酸多態(tài)性（SNP） 卵巢上皮性癌 鉑類化療 臨床預(yù)后　出處：《河北醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：卵巢惡性腫瘤是女性生殖系統(tǒng)常見的三大惡性腫瘤之一，其中最常見的婦科惡性腫瘤為上皮性卵巢癌（epithelial ovarian cancer，EOC)，它在導(dǎo)致女性死亡的病因中排名第七位，患者5年生存率約30%[1]。EOC患者的治療主要包括手術(shù)和術(shù)后的輔助化療。以鉑類為基礎(chǔ)的聯(lián)合化療是EOC患者術(shù)后的一線治療方案，但患者對鉑類的耐藥嚴(yán)重影響了化療療效，成為治療EOC的一大障礙。鉑類耐藥可能受累于多種機(jī)制，個體遺傳因素所導(dǎo)致的對藥物的反應(yīng)差異在治療中起著重要作用。 P-糖蛋白是一種膜轉(zhuǎn)運(yùn)蛋白，它位于細(xì)胞漿膜，可將細(xì)胞內(nèi)的化療藥物泵出細(xì)胞外，使細(xì)胞內(nèi)藥物濃度下降，增加了腫瘤細(xì)胞的耐藥性。P-糖蛋白表達(dá)水平的高低可能導(dǎo)致上皮性卵巢癌患者對鉑類藥物的應(yīng)答和預(yù)后的差異，而編碼P-糖蛋白的基因即為多藥耐藥基因1（multidrugresistance1，MDR1）。同樣，細(xì)胞對藥物代謝解毒功能的改變亦與鉑類耐藥密切相關(guān)。鉑類藥物進(jìn)入細(xì)胞后在谷胱甘肽-S-轉(zhuǎn)移酶P1（glutathione-S-transferases P1, GSTP1）的催化下可與谷胱甘肽（glutathione,GSH）形成復(fù)合物，該復(fù)合物毒性降低且易于被轉(zhuǎn)運(yùn)出細(xì)胞，有利于鉑類藥物從尿液或膽汁中排出，從而達(dá)到代謝解毒的功能[2]。位于MDR1、GSTP1基因的遺傳變異可能通過改變其所編碼蛋白質(zhì)的表達(dá)，進(jìn)而影響患者對鉑類藥物的應(yīng)答及臨床預(yù)后。 本研究擬探討MDR1和GSTP1基因遺傳變異與上皮性卵巢癌患者鉑類耐藥及臨床預(yù)后的關(guān)系。旨在尋找上皮性卵巢癌患者鉑類化療的分子標(biāo)志物，為臨床的“個體化”治療提供重要的實(shí)驗(yàn)數(shù)據(jù)。 方法：采用回顧性病例研究方法，病例為2001年12月至2010年6月期間在河北醫(yī)科大學(xué)第四醫(yī)院婦科進(jìn)行治療的上皮性卵巢癌患者（225例）。術(shù)前采集外周靜脈血5ml，以蛋白酶K消化-飽和氯化鈉鹽析法提取基因組DNA，采用連接酶檢測反應(yīng)（LDR）檢測上皮性卵巢癌患者的MDR1基因C3435T、G2677T/A和GSTP1基因A313G這3個SNP位點(diǎn)的基因型和等位基因頻率分布情況。所有患者均經(jīng)過3年隨訪。判定患者臨床預(yù)后的指標(biāo)包括患者對鉑類化療的應(yīng)答、總生存時間（overallsurvival，OS）和疾病無進(jìn)展生存期（progression-free survival，PFS）。 數(shù)據(jù)分析采用SPSS13.0統(tǒng)計軟件進(jìn)行處理。采用χ2檢驗(yàn)檢測基因型和等位基因與鉑類化療應(yīng)答的關(guān)系；以logistic regression方法計算相對風(fēng)險度的比值比(odds ratio, OR)及其95%可信區(qū)間(confidence interval, CI)；生存分析采用Kaplan-Meier法，各因素水平間比較用Log-rank及Breslow分析，Cox回歸模型進(jìn)一步分析預(yù)后相關(guān)因素（患者年齡、腫瘤分化程度、病理類型、臨床分期、殘余病灶）。P＜0.05作為差異有顯著性的標(biāo)準(zhǔn)。 結(jié)果： 1MDR1基因C3435T、G2677T/A和GSTP1基因A313G三個多態(tài)位點(diǎn)的基因型和等位基因頻率分布在鉑類化療應(yīng)答組與不應(yīng)答組之間均無統(tǒng)計學(xué)差異（P＞0.05）。 2MDR1基因C3435T與上皮性卵巢癌患者臨床生存預(yù)后的關(guān)系：患者以復(fù)發(fā)組和未復(fù)發(fā)組分類進(jìn)行生存分析，C/C、C/T、T/T三種基因型在復(fù)發(fā)組和未復(fù)發(fā)組的頻率分布分別為35.8%、47.0%、17.2%和26.4%、57.1%、16.5%，經(jīng)Kaplan-Meier單因素分析兩組比較無統(tǒng)計學(xué)差異（P0.05），，進(jìn)一步的Cox回歸多因素分析亦無統(tǒng)計學(xué)差異（P0.05）；以生存組和死亡組分類，三種基因型頻率分布分別為31.6%、52.9%、15.4%和32.6%、48.3%、19.1%， Kaplan-Meier單因素分析和Cox回歸多因素分析顯示均無統(tǒng)計學(xué)差異（P0.05）。 3MDR1基因G2677T/A與上皮性卵巢癌患者臨床生存預(yù)后的關(guān)系：G/G、G/T、A/G、A/A、A/T、T/T六種基因型在復(fù)發(fā)組和未復(fù)發(fā)組的頻率分布分別為：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基因型進(jìn)一步分類，在復(fù)發(fā)組和未復(fù)發(fā)組的頻率分布分別為：17.9%、82.1%和23.1%、76.9%。Cox回歸多因素分析，復(fù)發(fā)組和未復(fù)發(fā)組比較具有統(tǒng)計學(xué)意義（P=0.01，HR=3.27，95%CI=1.36-7.88）。以生存組和死亡組分類，頻率分布分別為：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%，經(jīng)Kaplan-Meier單因素分析和Cox回歸多因素分析，6種基因型的頻率分布無統(tǒng)計學(xué)差異。 4GSTP1基因A313G與上皮性卵巢癌患者臨床生存預(yù)后的關(guān)系：患者以復(fù)發(fā)組和未復(fù)發(fā)組分類進(jìn)行生存分析， A/A、A/G、G/G三種基因型的頻率分布分別為：56.7%、38.1%、5.2%和64.8%、31.9%、3.3%，以Kaplan-Meier單因素分析和Cox回歸多因素分析均無統(tǒng)計學(xué)差異（P0.05）；三種基因型在生存組和死亡組的頻率分布分別為60.3%、37.5%、2.2%和59.6%、32.6%、7.9%，以Kaplan-Meier單因素分析和Cox回歸多因素分析比較亦無統(tǒng)計學(xué)差異（P0.05）。 結(jié)論： 1MDR1基因C3435T、G2677T/A和GSTP1基因A313G多態(tài)可能與上皮性卵巢癌患者鉑類化療應(yīng)答無關(guān)。 2MDR1基因C3435T多態(tài)可能與上皮性卵巢癌患者鉑類化療后的復(fù)發(fā)和生存時間無關(guān)。 3MDR1基因G2677T/A多態(tài)性可能與中國北方婦女上皮性卵巢癌患者鉑類化療后的復(fù)發(fā)相關(guān)，但與其生存時間無關(guān)。 4GSTP1基因A313G多態(tài)可能與上皮性卵巢癌鉑類化療患者的復(fù)發(fā)和生存時間無關(guān)。
[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.
Result錛

本文編號：1445466