本文選題：肺腺癌 + 免疫組化　； 參考：《皖南醫(yī)學院》2017年碩士論文

【摘要】：目的:尋找一種適合于肺腺癌間變淋巴瘤激酶(anaplastic lymphoma kinase,ALK)融合基因檢測的初篩方法,探討肺腺癌間變淋巴瘤融合基因陽性率并分析其與臨床特征的關(guān)系。方法:收集整理皖南醫(yī)學院弋磯山醫(yī)院病理科2014.1.1~2016.3.1日期間行病理檢查且病理結(jié)果為肺腺癌并均已行ALK常規(guī)免疫組織化學(immunohistochemistry,IHC)檢查(一抗試劑采用1A4高親和力抗體)病例181例(包括手術(shù)標本54例、CT引導下經(jīng)皮肺穿刺活檢或支氣管鏡活檢標本122例、細胞蠟塊標本5例)。試驗分兩部分,第一部分對54例手術(shù)標本先制作組織芯片,而后行熒光原位雜交(fluorescence in situ hybridization,FISH)檢查,以FISH作為ALK重排基因檢測標準,分析手術(shù)標本免疫組化ALK(1A4)的敏感度和特異度,決定其是否適合作為肺腺癌ALK融合基因檢測初篩方法。第二部分對所有患者ALK(1A4)結(jié)果為1+、2+、3+患者行FISH檢查,分析肺腺癌患者ALK陽性率并分析影響其陽性率的相關(guān)因素。實驗結(jié)果使用SPSS16.0統(tǒng)計軟件中的χ2檢驗處理,分析ALK陽性率與年齡、是否轉(zhuǎn)移、性別、是否抽煙、腫瘤臨床分期、組織分化的相關(guān)性,當n≥40,1≤T5,需使用連續(xù)校正公式計算χ2值,當n40或T1時,使用Fisher確切概率法分析,均以P0.05認為有統(tǒng)計學意義。結(jié)果:已行ALK(1A4)檢測的肺腺癌手術(shù)標本54例患者,其中男性31例,女性23例,男女比例為1.35:1;患者年齡最大88歲,最小32歲,平均年齡60.4歲;38例為Ⅰ期患者,13例為Ⅱ期患者,2例為Ⅲ期患者,僅1例為Ⅳ期患者;低分化21例,中分化32例,高分化僅1例;19例有吸煙史,35例無吸煙史。檢測結(jié)果顯示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例患者中未檢測到ALK(1A4)3+病例;其余45例為ALK(1A4)-患者,FISH結(jié)果均為陰性。FISH檢測結(jié)果顯示手術(shù)病例肺腺癌ALK陽性率為9.3%。ALK(1A4)抗體敏感度為100%,特異度為91.8%。本實驗所收集的181(包括手術(shù)標本54例、CT引導下經(jīng)皮肺穿刺活檢或支氣管鏡活檢標本122例、細胞蠟塊標本5例)例患者。103例男性患者共檢出9例ALK基因重排陽性患者,重排發(fā)生率為8.7%,78例女性患者共檢出11例ALK重排陽性患者,重排發(fā)生率為14.1%,兩者比較結(jié)果并無統(tǒng)計學差異(P=0.2543)。入組患者年齡60歲的是65例,ALK基因重排陽性者12例,重排發(fā)生率為18.5%;≥60歲者為116例,結(jié)果8例為ALK重排陽性者,重排發(fā)生率為6.9%,兩者比較有統(tǒng)計學差異(P=0.0173)。入組患者中有128例有吸煙史,ALK基因重排陽性者9例,重排發(fā)生率為7.0%,53例患者無吸煙史,11例為ALK重排陽性者,重排發(fā)生率為20.8%,兩者比較有統(tǒng)計學差異(P=0.0074)。57例出現(xiàn)轉(zhuǎn)移的患者共檢出4例ALK基因重排陽性患者,重排發(fā)生率為7.0%,124例無轉(zhuǎn)移的患者共檢出16例ALK重排陽性患者,重排發(fā)生率為12.9%,兩者比較并無統(tǒng)計學差異(P=0.2407)。63例低分化腺癌患者,共檢出3例ALK基因重排陽性患者,重排發(fā)生率為4.8%;115例中分化腺癌患者,共檢出17例ALK重排陽性患者,重排發(fā)生率為14.8%;3例高分化腺癌患者,沒有檢出ALK重排陽性患者,三者比較并無統(tǒng)計學差異(P=0.1035)。Ⅰ期患者52例,共檢出7例ALK基因重排陽性患者,重排發(fā)生率為13.5%;Ⅱ期患者34例,共檢出6例ALK重排陽性患者,重排發(fā)生率為17.6%;Ⅲ期患者38例,共檢出3例ALK基因重排陽性患者,重排發(fā)生率為7.9%;Ⅳ期患者57例,共檢出4例ALK基因重排陽性患者,重排發(fā)生率為7.0%,四者比較并無統(tǒng)計學差異(P=0.3704)。ALK(1A4)檢測結(jié)果為1+、2+、3+患者共計36例,其中有9例是手術(shù)標本,已做過FISH檢測(5例ALK陽性),在這里就不重復做FISH檢測。接下來的試驗需要對27例ALK(1A4)檢測結(jié)果為1+、2+、3+患者進行FISH檢測,結(jié)果顯示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(-);未檢測到ALK(1A4)3+病例。最終FISH檢測結(jié)果陽性患者共20例,陽性率為11.0%。結(jié)論:1、54例已行ALK(1A4)抗體檢測的肺腺癌手術(shù)標本進行FISH檢測結(jié)果顯示,ALK(1A4)抗體敏感度為100%,特異度為91.8%,同時其敏感度、特異度與2013版ALK陽性NSCLC診斷專家共識推薦的5A4、D5F3抗體相似,因此ALK(1A4)抗體可以作為ALK融合基因陽性肺腺癌的初篩方法。2、181例患者FISH檢測結(jié)果ALK重排陽性患者共20例,陽性率為11.0%,ALK基因重排多發(fā)生于不吸煙、較年輕(60歲)的患者,而性別、腫瘤細胞分化、有無轉(zhuǎn)移、臨床分期與ALK基因重排無明顯統(tǒng)計學差異(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).
【學位授予單位】：皖南醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R734.2

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