荧光原位杂交技术检测非小细胞肺癌患者现场细胞学制片表皮生长因子受体基因扩增状态的研究

发布时间：2018-06-28 07:46

本文选题：非小细胞肺癌 + 荧光原位杂交　；参考：《天津医科大学》2015年硕士论文

【摘要】：研究目的本研究创新性应用荧光原位杂交(FISH)技术检测非小细胞肺癌(NSCLC)患者现场细胞学(ROSE)制片的表皮生长因子受体(EGFR)基因扩增状态,旨在探讨以NSCLC患者气管镜取得细胞学或组织块标本的ROSE制片为标本,FISH检测其EGFR基因扩增状态的应用。以及探讨EGFR基因扩增状态在NSCLC患者中与其年龄、性别、吸烟状态、病理类型、分化程度、肿瘤TNM分期等临床特征的相关性。研究方法本研究在2013年9月至2014年12月期间对就诊于天津医科大学总医院呼吸内科气管镜室且胸部X线或CT检查怀疑肺部恶性肿瘤性病变而行诊断性治疗的患者在气管镜检查过程中予以ROSE制片指导其进行程度。然后应用FISH技术在原ROSE制片上检测71例最终取材经过病理科医师细胞学或组织学确诊为NSCLC患者的EGFR基因扩增状态,并将FISH结果与患者临床特征进行统计学比较。其数据结果应用SPSS16.0统计软件处理,P0.05视为差异具有统计学意义。结果1.在本研究中ROSE恶性细胞阳性并且考虑NSCLC的72例患者仅有3例因取材量小而未得到病理科确诊,病理报告1例为未找到肿瘤细胞,另外2例为可见可疑核异质细胞;另有2例现场ROSE恶性细胞阳性但观察形态考虑为小细胞肺癌(SCLC)的患者最终经病理科确诊病理类型为低分化鳞癌;本研究中ROSE对NSCLC患者制片中恶性细胞的现场诊断与最终病理确诊的符合率可达95.9%,且研究进行期间ROSE诊断未出现假阴性的结果;最终共搜集NSCLC患者ROSE制片71例行FISH检测,其中有68例(95.8%)患者顺利取得结果,另外3例患者ROSE制片分别因为1例细胞数目过少、1例细胞重叠过多影响信号读取与1例杂交后出现部分区域无杂交信号及部分区域杂交率较低而导致FISH检测的失败。2.其EGFR基因发生扩增,即FISH(+)的患者共23例,其中7例为簇状扩增。EGFR基因扩增率为33.8%。并且经过统计学分析,EGFR基因扩增状态与患者性别(χ2=0.176,P=0.675)、年龄(χ2=2.796,P=0.094)、吸烟状态(χ2=0.584,P=0.445)、病理类型(χ2=1.248,P=0.264)、肿瘤分化程度(χ2=1.234,P=0.267)、肿瘤TNM分期(P=0.474)的统计结果均无明显统计学差异(P均0.05)。结论本研究搜集NSCLC患者ROSE制片,创新性的应用FISH技术在ROSE制片的原片上进行EGFR基因扩增的检测,研究结果表明:1.NSCLC患者气管镜细胞学及组织块标本的ROSE阅片结果与最终诊断结果一致性良好,并且FISH技术可应用于ROSE制片作为标本来检测EGFR基因的扩增状态,此方法不仅使得细胞学的FISH研究更加科学、严谨;而且较传统方法更加快捷、经济。但应注意提高气管镜室ROSE制片的质量,因为其质量可直接影响气管镜检查的进度及FISH结果的取得。FISH结果的顺利取得还需在进行过程中注意杂交区域的正确选择、水浴与消化等步骤的适度等。2.本研究中NSCLC患者的EGFR基因扩增率为33.8%,且其在不同性别、不同年龄、不同吸烟状态、不同病理类型、不同肿瘤分化程度、不同TNM分期中不存在明显的差异,即未发现NSCLC患者的EGFR基因扩增状态与临床特征的显著相关性。
[Abstract]:The purpose of this study was to detect the amplification state of epidermal growth factor receptor (EGFR) gene in the field cytology (ROSE) of non small cell lung cancer (NSCLC) patients by innovative application of fluorescence in situ hybridization (FISH). The purpose of this study was to explore the ROSE filmmaking of cytology or tissue specimens obtained from the trachea of NSCLC patients as specimens. FISH was used to detect the expansion of the EGFR gene. The correlation of EGFR gene amplification status in NSCLC patients with age, sex, smoking status, pathological type, degree of differentiation, TNM staging, and other clinical features. The study was conducted in the trachea room of the respiratory department of General Hospital Affiliated to Tianjin Medical University during the period from September 2013 to December 2014. The X-ray or CT examination of the patients who suspected pulmonary malignant tumor and diagnostic treatment were guided by ROSE in the process of tracheal endoscopy. Then, the FISH technique was used to detect the EGFR gene amplification status of 71 cases in the original ROSE film, which were determined by the cytology or histology of the pathologist. The results of FISH were compared with the clinical features of the patients. The results of the data were treated with SPSS16.0 software, and P0.05 was statistically significant. Results 1. in this study, only 3 cases of ROSE malignant cells and 72 patients considering NSCLC were not confirmed by the pathology department, and 1 cases were not reported. The tumor cells were found, the other 2 cases were suspicious nuclear heterocells, and 2 cases of ROSE malignant cells were positive but the patients with small cell lung cancer (SCLC) were diagnosed as a low differentiated squamous cell carcinoma. In this study, the field diagnosis and final pathological diagnosis of the malignant cells in the NSCLC patients were in the field and in the final pathological diagnosis. The coincidence rate reached 95.9%, and the results of ROSE diagnosis during the study did not appear false negative; finally, a total of 71 cases of ROSE production in NSCLC patients were detected by FISH, of which 68 cases (95.8%) were successfully obtained, and the other 3 cases of ROSE produced 1 cases with small number of cells, 1 cells overlapped overoverlapping signal reading and 1 hybrids. There were no hybridization signals in some regions and low hybridization rates in some regions that resulted in the failure of FISH detection in.2.. The EGFR gene was amplified, that is, 23 cases of FISH (+), of which 7 cases were amplified by cluster amplification of.EGFR gene amplification rate of 33.8%. and statistically analyzed, EGFR gene expansion and patient sex (chi 2=0.176, P=0.675), age (x 2=2). .796, P=0.094), smoking status (x 2=0.584, P=0.445), pathological type (x 2=1.248, P=0.264), the degree of tumor differentiation (2=1.234, P=0.267), and the statistical results of the tumor TNM staging (P=0.474) were not statistically significant (P all 0.05). The results of gene amplification test showed that the results of ROSE scanning in trachea and tissue specimens of 1.NSCLC patients were in good agreement with the final diagnosis, and the FISH technique could be used as a specimen to detect the amplification state of EGFR gene. This method not only made the FISH research of cytology more scientific and rigorous. It is more rapid and economical than the traditional method. However, attention should be paid to improving the quality of ROSE filmmaking in the trachea room, because the quality of the trachea can directly affect the progress of the trachea and the results of the FISH result in the smooth acquisition of the.FISH results. It is necessary to pay attention to the correct selection of the hybridization area in the process, and in the moderate.2. study of the steps of water bath and digestion. The amplification rate of EGFR gene in NSCLC patients was 33.8%, and there was no significant difference in different sex, different age, different smoking status, different pathological types, different degree of differentiation of tumor and different TNM staging, that is, no significant correlation was found between the amplification state of EGFR gene and clinical characteristics in NSCLC patients.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R734.2

【共引文献】