肺腺癌ALK基因检测及与患者临床特征关系

发布时间：2018-06-23 05:21

本文选题：肺腺癌 + 免疫组化　；参考：《皖南医学院》2017年硕士论文

【摘要】：目的:寻找一种适合于肺腺癌间变淋巴瘤激酶(anaplastic lymphoma kinase,ALK)融合基因检测的初筛方法,探讨肺腺癌间变淋巴瘤融合基因阳性率并分析其与临床特征的关系。方法:收集整理皖南医学院弋矶山医院病理科2014.1.1~2016.3.1日期间行病理检查且病理结果为肺腺癌并均已行ALK常规免疫组织化学(immunohistochemistry,IHC)检查(一抗试剂采用1A4高亲和力抗体)病例181例(包括手术标本54例、CT引导下经皮肺穿刺活检或支气管镜活检标本122例、细胞蜡块标本5例)。试验分两部分,第一部分对54例手术标本先制作组织芯片,而后行荧光原位杂交(fluorescence in situ hybridization,FISH)检查,以FISH作为ALK重排基因检测标准,分析手术标本免疫组化ALK(1A4)的敏感度和特异度,决定其是否适合作为肺腺癌ALK融合基因检测初筛方法。第二部分对所有患者ALK(1A4)结果为1+、2+、3+患者行FISH检查,分析肺腺癌患者ALK阳性率并分析影响其阳性率的相关因素。实验结果使用SPSS16.0统计软件中的χ2检验处理,分析ALK阳性率与年龄、是否转移、性别、是否抽烟、肿瘤临床分期、组织分化的相关性,当n≥40,1≤T5,需使用连续校正公式计算χ2值,当n40或T1时,使用Fisher确切概率法分析,均以P0.05认为有统计学意义。结果:已行ALK(1A4)检测的肺腺癌手术标本54例患者,其中男性31例,女性23例,男女比例为1.35:1;患者年龄最大88岁,最小32岁,平均年龄60.4岁;38例为Ⅰ期患者,13例为Ⅱ期患者,2例为Ⅲ期患者,仅1例为Ⅳ期患者;低分化21例,中分化32例,高分化仅1例;19例有吸烟史,35例无吸烟史。检测结果显示ALK(1A4)1+患者共6例,其中3例FISH(+),3例FISH(-);ALK(1A4)2+患者共3例,其中2例FISH(+),1例患者FISH(-);54例患者中未检测到ALK(1A4)3+病例;其余45例为ALK(1A4)-患者,FISH结果均为阴性。FISH检测结果显示手术病例肺腺癌ALK阳性率为9.3%。ALK(1A4)抗体敏感度为100%,特异度为91.8%。本实验所收集的181(包括手术标本54例、CT引导下经皮肺穿刺活检或支气管镜活检标本122例、细胞蜡块标本5例)例患者。103例男性患者共检出9例ALK基因重排阳性患者,重排发生率为8.7%,78例女性患者共检出11例ALK重排阳性患者,重排发生率为14.1%,两者比较结果并无统计学差异(P=0.2543)。入组患者年龄60岁的是65例,ALK基因重排阳性者12例,重排发生率为18.5%;≥60岁者为116例,结果8例为ALK重排阳性者,重排发生率为6.9%,两者比较有统计学差异(P=0.0173)。入组患者中有128例有吸烟史,ALK基因重排阳性者9例,重排发生率为7.0%,53例患者无吸烟史,11例为ALK重排阳性者,重排发生率为20.8%,两者比较有统计学差异(P=0.0074)。57例出现转移的患者共检出4例ALK基因重排阳性患者,重排发生率为7.0%,124例无转移的患者共检出16例ALK重排阳性患者,重排发生率为12.9%,两者比较并无统计学差异(P=0.2407)。63例低分化腺癌患者,共检出3例ALK基因重排阳性患者,重排发生率为4.8%;115例中分化腺癌患者,共检出17例ALK重排阳性患者,重排发生率为14.8%;3例高分化腺癌患者,没有检出ALK重排阳性患者,三者比较并无统计学差异(P=0.1035)。Ⅰ期患者52例,共检出7例ALK基因重排阳性患者,重排发生率为13.5%;Ⅱ期患者34例,共检出6例ALK重排阳性患者,重排发生率为17.6%;Ⅲ期患者38例,共检出3例ALK基因重排阳性患者,重排发生率为7.9%;Ⅳ期患者57例,共检出4例ALK基因重排阳性患者,重排发生率为7.0%,四者比较并无统计学差异(P=0.3704)。ALK(1A4)检测结果为1+、2+、3+患者共计36例,其中有9例是手术标本,已做过FISH检测(5例ALK阳性),在这里就不重复做FISH检测。接下来的试验需要对27例ALK(1A4)检测结果为1+、2+、3+患者进行FISH检测,结果显示ALK(1A4)1+患者共17例,其中8例FISH(+),9例FISH(-);ALK(1A4)2+患者共10例,其中7例FISH(+),3例患者FISH(-);未检测到ALK(1A4)3+病例。最终FISH检测结果阳性患者共20例,阳性率为11.0%。结论:1、54例已行ALK(1A4)抗体检测的肺腺癌手术标本进行FISH检测结果显示,ALK(1A4)抗体敏感度为100%,特异度为91.8%,同时其敏感度、特异度与2013版ALK阳性NSCLC诊断专家共识推荐的5A4、D5F3抗体相似,因此ALK(1A4)抗体可以作为ALK融合基因阳性肺腺癌的初筛方法。2、181例患者FISH检测结果ALK重排阳性患者共20例,阳性率为11.0%,ALK基因重排多发生于不吸烟、较年轻(60岁)的患者,而性别、肿瘤细胞分化、有无转移、临床分期与ALK基因重排无明显统计学差异(P0.05)。
[Abstract]:Objective: to find a preliminary screening method suitable for the detection of anaplastic lymphoma kinase (ALK) fusion gene fusion gene, explore the positive rate of the fusion gene of the pulmonary adenocarcinoma cell lymphoma and analyze its relationship with the clinical characteristics. Methods: collect and collate the 2014.1.1~2016.3.1 days of the pathology department of the Yi La hospital of Wangnan Medical College. Pathological examination and pathological results were pulmonary adenocarcinoma and 181 cases (including 54 cases of surgical specimens, 122 cases of CT guided percutaneous lung biopsy or bronchoscopic biopsy, 5 cases of cell wax block) had been examined by ALK routine immunohistochemical (immunohistochemistry, IHC) examination (an anti reagents with 1A4 high affinity antibody). The test was divided into two parts. In the first part, the tissue chips were made in 54 cases, and the fluorescence in situ hybridization (FISH) was examined by fluorescence in situ hybridization. FISH was used as the ALK rearrangement gene detection standard, and the sensitivity and specificity of ALK (1A4) were analyzed for the immune histochemical ALK (1A4) of the surgical specimens, and it was decided whether it was suitable as a screening prescription for the detection of ALK fusion gene for lung adenocarcinoma. Method. The second part of all patients ALK (1A4) results for 1+, 2+, 3+ patients with FISH examination, analysis of ALK positive rate in patients with lung adenocarcinoma and analysis of the correlation factors affecting the positive rate. The experimental results were treated with the x 2 test in SPSS16.0 statistics software to analyze the positive rate and age of ALK, whether metastasis, sex, smoking, tumor clinical staging, tissue The correlation of differentiation, when n > 40,1 < < T5 >, we need to use the continuous correction formula to calculate the x 2 value. When N40 or T1, the exact probability method of Fisher is used to analyze it with P0.05. Results: there are 54 patients with ALK (1A4) detection of lung adenocarcinoma, including 31 males and 23 females, and the male and female ratio is 1.35:1; the age of the patient is 88 years old. The minimum 32 years, the average age of 60.4 years, 38 cases of stage I patients, 13 cases of stage II patients, 2 cases of stage III patients, only 1 cases of stage IV patients, 21 cases of low differentiation, 32 cases of differentiation, 1 cases of high differentiation, 19 cases of smoking history and 35 non smoking history. The results showed that ALK (1A4) 1+ patients were 6 cases, 3 cases FISH (+), FISH (-); ALK (1A4) 2+ patients, There were 2 cases of FISH (+) and 1 patients with FISH (-); 54 cases were not detected ALK (1A4) 3+ cases; the other 45 cases were ALK (1A4) - patients and FISH results were negative.FISH detection results showed that the ALK positive rate of lung adenocarcinoma was 100%, and the specificity was 181 (including 54 cases of surgical specimens. Guided by percutaneous lung biopsy or bronchoscopic biopsy specimens of 122 cases, 5 cases of cell wax block specimens, 9 cases of.103 male patients were detected by ALK gene rearrangement positive, the incidence of rearrangement was 8.7%. 11 cases of ALK rearrangement positive patients were detected in 78 cases, and the incidence of rearrangement was 14.1%, and there was no statistical difference between the two cases (P=0.25 43). There were 65 patients aged 60 years old, 12 cases of ALK gene rearrangement, 18.5% for rearrangement, 116 for 60 years old, and 8 with positive rearrangement, and the rate of rearrangement was 6.9% (P=0.0173). 128 cases had history of smoking, 9 cases of ALK gene rearrangement positive, and recurrence rate of 7%, in the group of patients. No smoking history in 53 cases, 11 cases of ALK rearrangement positive, the incidence of rearrangement was 20.8%. There were statistically significant differences (P=0.0074) in the patients with.57 cases, 4 cases of ALK gene rearrangement positive patients were detected, the incidence of rearrangement was 7%, and 16 cases of ALK rearrangement positive patients were detected in 124 cases without metastasis. The incidence of rearrangement was 12.9%, both ratio was compared. There was no statistical difference (P=0.2407) in.63 patients with low differentiated adenocarcinoma, 3 cases of ALK gene rearrangement positive patients were detected, the incidence of rearrangement was 4.8%. In 115 cases of differentiated adenocarcinoma, 17 cases of ALK rearrangement positive patients were detected, the incidence of rearrangement was 14.8%; 3 cases of highly differentiated adenocarcinoma were not detected by ALK rearrangement positive patients, and the three had no statistics. Study difference (P=0.1035). 52 cases of stage I patients, 7 cases of ALK gene rearrangement positive patients were detected, the incidence of rearrangement was 13.5%, 34 cases in stage II patients, 6 cases of ALK rearrangement positive, 17.6% of rearrangement, 38 cases in stage III patients, 3 cases of ALK gene rearrangement positive, the incidence of rearrangement was 7.9%, 57 cases in stage IV patients, a total of 4 cases of ALK base were detected. The recurrence rate of rearrangement positive patients was 7%, and there was no statistical difference between the four (P=0.3704) and 36 cases of 1+, 2+, 3+ patients, of which 9 were surgical specimens, and FISH detection (5 ALK positive) had been done (5 cases of ALK positive), and there was no repetition of FISH detection here. The next test needed to test the results of 27 ALK (1A4) for 1+, 3, 3. A total of 17 patients with ALK (1A4) 1+ were detected in 17 cases, including 8 cases of FISH (+), 9 cases of FISH (-) and 10 cases of ALK (1A4) 2+, of which 7 cases were FISH (+), 3 patients were FISH (-); 20 cases were not detected. The positive rate was the conclusion of the lung adenocarcinoma hand. The results of FISH detection showed that the sensitivity of ALK (1A4) antibody was 100% and the specificity was 91.8%, and its sensitivity, specificity and D5F3 antibody were similar to 5A4, which was recommended by 2013 ALK positive NSCLC diagnostic experts. Therefore, ALK (1A4) antibody could be used as a screening method for ALK fusion gene positive lung adenocarcinoma. The positive rate of rearrangement of positive patients was 20, the positive rate was 11%. The ALK gene rearrangement occurred mostly in non smoking and younger patients (60 years old), while sex, tumor cell differentiation, metastasis, clinical stage and ALK gene rearrangement had no significant difference (P0.05).
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2

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