基于上皮间质转化学说的乳腺癌循环肿瘤细胞异质性的探索性研究

发布时间：2018-05-19 21:26

  本文选题：乳腺癌 + 循环肿瘤细胞　； 参考：《北京协和医学院》2017年硕士论文

【摘要】：目的:既往大量研究表明循环肿瘤细胞(circulating tumor cells,CTCs)计数是转移性乳腺癌预后因子,但循环肿瘤细胞的异质性,如上皮间质转化可能会影响其临床应用价值。乳腺癌治疗方案的制定依赖于分子分型的指导,由于在侵袭和转移的过程中,肿瘤分子分型可能发生了一定程度的改变,而有时往往无法直接获得转移灶的病理组织,应用液态活检技术进行CTCs的雌/孕激素受体(ER/PR)检测可以实现对ER/PR状态的实时动态监测。本研究旨在探究基于上皮间质转化学说的CTCs分型与乳腺癌ER/PR状态的关系,探索从上皮间质转化和CTCs的ER/PR状态两方面兼顾CTCs异质性的简便易行的检测方法。方法:该研究纳入28名转移性乳腺癌患者,应用CanpatrolTM二代CTC检测技术进行外周血CTCs检测,采用多重RNA原位杂交的方法对富集的CTCs进行特异性基因核酸定位,将CTCs分为上皮型、间质型、混合型三种亚型。同时,通过与内参基因的荧光信号强度对比将每个CTC的ER/PR状态分为高表达、中表达、低表达、无表达四种状态。结果:原发灶ER/PR状态不同,患者外周血中检测到CTCs亚型由上皮型至间质型的分布差异有统计学意义(Z=-3.569,P0.001);原发灶ER/PR阳性的患者,其上皮型CTCs所占的比例(41.7%)高于混合型(27.4%)和间质型CTCs(30.9%);原发灶ER/PR阴性患者,间质型CTCs所占的比例(43.1%)高于上皮型(30.5%)和混合型CTCs(26.4%)。此外,原发灶ER/PR状态不同,转移后外周血CTCs的ER/PR状态由高表达至低表达的变化趋势也有明显差异(Z=-3.524,P0.001);原发灶ER/PR阳性的患者,其CTCs的ER/PR状态高表达所占的比例(5.8%)高于原发灶ER/PR阴性的患者(3.5%);原发灶ER/PR阴性的患者,其CTCs的ER/PR状态无表达所占的比例(35.0%)高于原发灶ER/PR阳性的患者(23.7%)。同时,不同CTCs亚型的ER/PR状态变化趋势不同(Z=6.405,P=0.041);上皮型CTCs中ER/PR状态高表达所占的比例(8.3%)高于混合型(2.1%)和间质型CTCs(3.2%)。结论:外周血CTCs亚型由上皮型至间质型的分布差异与原发灶ER/PR状态有关,CTCs的ER/PR状态由高表达至低表达的变化趋势也随着原发灶ER/PR状态不同而变化。通过实时监测不同亚型CTCs的ER/PR状态,可以为临床提供简便易行的CTCs检测方法用于指导晚期乳腺癌患者的内分泌治疗方案的选择。目的:既往研究表明晚期乳腺癌的侵袭和转移与上皮间质转化的过程相关。然而对于早期乳腺癌而言,基于上皮间质转化学说的循环肿瘤细胞(circulating tumor cells,CTCs)分型与其临床病理特征的关系仍不清楚。本研究旨在探究基于上皮间质转化学说的CTCs分型与早期乳腺癌的临床病理特征间的关系,以期为基于上皮间质转化学说的循环肿瘤细胞分型用于早期乳腺癌的预后判断提供参考。方法:本研究共纳入150名早期乳腺癌患者,应用CanpatrolTM二代CTC检测技术进行外周血CTCs检测,采用多重RNA原位杂交的方法对富集的CTCs进行特异性基因核酸定位,将CTCs分为上皮型、间质型、混合型三种亚型。结果:通过与患者的临床病理特征进行对比发现,患者的TNM分期不同,其外周血中检测到CTC亚型由上皮型至间质型的分布差异有统计学意义(Z= 39.723,P0.001),Ⅱ期(41.5%)和Ⅲ期(43.4%)的患者中检测出的间质型CTCs明显高于Ⅰ期患者(29.0%)。此外,在不同复发风险分组之间,其CTC亚型由上皮型至间质型的分布差异也有统计学意义(Z=-7.101,P0.001),高危组间质型CTCs所占的比例明显高于中危组(46.2%vs 32.6%)。而原发灶组织病理Ki 67水平的不同,CTCs各个亚型分布的变化趋势也存在差异(Z=-5.687,P0.001)。结论:对于早期乳腺癌患者,不同CTC亚型从上皮型到间质型的分布差异与其预后相关的临床病理特征有关,间质型CTCs所占比例较高时可能与一些预后不良的临床病理因素具有相似的提示意义,基于上皮间质转化学说的循环肿瘤细胞分型可以为早期乳腺癌的预后判断提供参考。
[Abstract]:Objective: a large number of previous studies have shown that the circulating tumor cells (CTCs) count is a prognostic factor for metastatic breast cancer, but the heterogeneity of circulating tumor cells, such as epithelial mesenchymal transition, may affect its clinical application. The formulation of breast cancer treatment options depends on the guidance of molecular typing and the invasion and metastasis In the process, the tumor molecular typing may change to a certain extent, and sometimes the pathological tissue of the metastases can not be obtained directly. The detection of the female / progesterone receptor (ER/PR) of CTCs by the liquid biopsy technique can realize the real-time dynamic monitoring of the state of the ER/PR. This study aims to explore the CTCs based on the theory of epithelial mesenchymal transition. The relationship between typing and ER/PR status of breast cancer, and exploring a simple and easy method for detecting CTCs heterogeneity from two aspects of epithelial mesenchymal transition and CTCs's ER/PR status. Methods: This study included 28 metastatic breast cancer patients, using the CanpatrolTM two generation CTC detection technique for peripheral blood CTCs detection and multiple RNA in situ hybridization. The specific gene nucleic acid location of the enriched CTCs was divided into three subtypes of epithelial, interstitial and mixed type. At the same time, the ER/PR state of each CTC was divided into high expression, medium expression, low expression, and no expression of four forms by contrast with the fluorescence intensity of the internal reference gene. Results: the ER/PR state of the primary foci was different and the peripheral blood was detected in the peripheral blood of the patients. The distribution of CTCs subtypes from epithelial to stromal type was statistically significant (Z=-3.569, P0.001), and the proportion of epithelial CTCs in primary ER/PR positive patients (41.7%) was higher than that of mixed type (27.4%) and interstitial CTCs (30.9%); the proportion of interstitial CTCs (43.1%) was higher than that of epithelial type (30.5%) and mixture of primary ER/PR negative patients (30.5%) and mixture. Type CTCs (26.4%). In addition, the ER/PR status of primary foci was different, and the ER/PR state of CTCs in peripheral blood was also significantly different from high expression to low expression (Z=-3.524, P0.001). The proportion of ER/PR state high expression of CTCs in the primary focal ER/PR positive patients (5.8%) was higher than that of the primary ER/PR negative (3.5%); the primary focal ER/PR was ER/PR. Negative patients, the proportion of CTCs ER/PR state non expression (35%) was higher than that of primary ER/PR positive patients (23.7%). Meanwhile, the change trend of ER/PR status in different CTCs subtypes was different (Z=6.405, P=0.041); the proportion of high expression of ER/PR in epithelial CTCs (8.3%) was higher than that of mixed type (2.1%) and interstitial CTCs (3.2%). Conclusion: peripheral blood (3.2%). The distribution of the blood CTCs subtypes from the epithelial type to the interstitial type is related to the ER/PR state of the primary focus. The trend of the ER/PR state from high expression to low expression of CTCs varies with the ER/PR state of the primary focus. By real-time monitoring of the ER/PR state of the different subtypes of CTCs, the simple and convenient CTCs detection method can be used for guidance in clinical practice. Endocrine therapy options for advanced breast cancer. Objective: Previous studies have shown that the invasion and metastasis of advanced breast cancer are associated with the process of epithelial mesenchymal transition. However, for early breast cancer, the circulating tumor cells (CTCs) classification based on the theory of epithelial mesenchymal transformation and its clinicopathological features The relationship is still unclear. The purpose of this study was to explore the relationship between CTCs typing based on epithelial mesenchymal transition and the clinicopathological features of early breast cancer in order to provide reference for predicting the prognosis of early breast cancer based on the theory of epithelial mesenchymal transition. Square method: This study included 150 early breast cancer patients. The CanpatrolTM two generation CTC detection technique was used to detect the peripheral blood CTCs, and the specific gene nucleic acid location of the enriched CTCs was carried out by multiple RNA in situ hybridization. The CTCs was divided into epithelial, interstitial and mixed subtypes. The results were compared with the clinical characteristics of the patients, and the patients' TNM stages were different. In peripheral blood, the distribution of CTC subtypes from epithelial to stromal type was statistically significant (Z= 39.723, P0.001). The interstitial CTCs detected in patients with stage II (41.5%) and stage III (43.4%) was significantly higher than that in phase I patients (29%). In addition, the distribution of CTC subtypes from epithelial to stromal types was also different between different recurrence risk groups. There were statistical significance (Z=-7.101, P0.001), and the proportion of interstitial CTCs in high-risk group was significantly higher than that in middle risk group (46.2%vs 32.6%). The variation trend of CTCs subtype distribution was also different (Z=-5.687, P0.001) in primary pathological Ki 67 levels (Z=-5.687, P0.001). Conclusion: for early breast cancer patients, different CTC subtypes from epithelial to interstitial type The distribution difference is related to the clinicopathological features related to the prognosis. The high proportion of interstitial CTCs may be of similar suggestive significance to some clinicopathological factors that have poor prognosis. The circulating tumor cell classification based on the theory of epithelial mesenchymal transformation can provide reference for the prognosis of early breast cancer.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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