Detection Of ALK Gene In Lung Adenocarcinoma And Its Relationship With Clinical Features

Objective: to find a suitable for lung adenocarcinoma anaplastic lymphoma kinase(anaplastic lymphoma kinase,ALK)detection of fusion gene screening method,to investigate the lung adenocarcinoma positive rate of anaplastic lymphoma fusion gene and analyzed its relationship with clinical characteristics.Methods: We collected the yijishan hospital of Wannan Medical College Department of pathology during 2014.1.1~2016.3.1 days for pathological examination and pathological results for lung cancer underwent routine ALK and immunohistochemical examination(immunohistochemistry,IHC)(an anti reagent using 1A4 high affinity antibody)patients181 cases(including 54 cases of surgical specimens,guided by CT percutaneous lung puncture biopsy or bronchoscopy biopsy specimens of 122 cases,cell block specimens of5 cases).The experiment was divided into two parts,the first part of the 54 cases of surgical specimens made of tissue microarray.Fluorescence in situ hybridization(fluorescence in situ hybridization,FISH)examination,with FISH as the detection of ALK gene rearrangement,ALK analysis of surgical specimens of immune group(1A4)sensitivity and specificity,determine its suitability as lung cancer screening method for detection of ALK fusion gene.The second part of all patients with ALK(1A4)results for1+,2+,3+ patients with FISH examination,analysis of lung adenocarcinoma patients with ALK positive rate,and analysis of the positive factors related to the positive rate.The experimental results using SPSS16.0 statistical software in the ?2 test,analysis of ALK positive rate and age,metastasis,gender,correlation between smoking,tumor staging,histological differentiation,When n ?40,1?T<5,need to use the Yates' continuitycorrected chi-square test,when n<40 or T <1,analyzed using Fisher's exact test,with P <0.05 was considered statistically significant.Results: ALK(1A4)in lung adenocarcinoma specimens was detected in 54 patients,including 31 cases of male,female 23 cases,male to female ratio was 1.35:1;the patients up to 88 years of age,the youngest was 32 years old,the average age of 60.4 years;38cases of stage I patients,13 cases of stage II patients,2 cases for stage III patients,only 1cases of stage IV patients;21 cases of low differentiated,moderately differentiated in 32 cases,only 1 cases of high differentiation;19 cases had a history of smoking,35 patients without smoking history.Test results show that ALK(1A4)1+ patients in 6 cases,including 3 cases of FISH(+),3 cases of FISH(-);ALK(1A4)2+ patients in 3 cases,including 2 cases of FISH(+),1 cases of patients with FISH(-);54 cases were not detected in ALK(1A4)3+ cases;the other 45 cases of ALK(1A4)-patients,FISH results were negative.FISH detection showed that the positive rate of ALK in lung adenocarcinoma was 9.3%.The sensitivity of ALK(1A4)antibody was 100% and specificity was 91.8%.In this study,181 were collected(including surgical specimens of 54 cases,CT guided percutaneous lung biopsy or bronchoscopic biopsy specimens in 122 cases,and 5 cases of cell paraffin specimens).103 cases of male patients were detected in 9 cases of patients with positive ALK gene rearrangement,rearrangement rate was 8.7%,78 cases of female patients were detected in 11 cases of patients with positive ALK rearrangement rearrangement,the incidence rate of 14.1%,the comparison between the two results were not statistically significant(P=0.2543).The patients under 60 years of age is 65 cases,12 cases of ALK gene rearrangement,rearrangement rate is 18.5%;more than 60 years old were 116 cases,8 cases for the ALK rearrangement,rearrangement rate was 6.9%,the difference was statistically significant(P=0.0173).The patients in 128 cases with a history of smoking,9 cases of ALK gene rearrangement,rearrangement rate was 7%,53 cases of patients with no smoking history,11 cases with ALK rearrangement,rearrangement rate was 20.8%,the difference was statistically significant(P=0.0074).57 cases of patients with metastasis were detected in 4 cases of ALK gene rearrangement positive patients,rearrangement rate was 7%,124 cases of patients without metastasis were detected in 16 cases of patients with positive ALK rearrangement rearrangement,the incidence rate of 12.9%,compared with no significant difference(P=0.2407).63 cases of low differentiated adenocarcinoma patients,3 cases were found in patients with positiveALK gene rearrangement,rearrangement of the incidence of 4.8%;patients with differentiated adenocarcinoma in 115 cases,17 cases were detected with ALK rearrangement positive patients,rearrangement rate is 14.8%;3 cases of high differentiated adenocarcinoma patients with no detectable ALK rearrangement positive patients,three there was no statistical difference(P=0.1035).52 cases of patients,7 cases were found in patients with positive ALK gene rearrangement,rearrangement rate is13.5%;34 cases of patients,6 cases were detected with ALK rearrangement rearrangement positive patients,the incidence of 17.6%;38 cases of stage III patients,there were 3 cases of patients with positive ALK gene rearrangement,rearrangement of the incidence rate was 7.9%;57 cases of stage IV patients,there were 4 cases of patients with positive ALK gene rearrangement,rearrangement rate was 7%,there was no statistical difference among the four(P=0.3704).ALK(1A4)test results for 1+,2+ and3+ patients,a total of 36 cases,including 9 cases of surgical specimens,have been tested by FISH(5 cases of ALK positive),here do not repeat the FISH test.The next test in 27 cases of ALK(1A4)1+,2+,test results for 3+ patients were detected by FISH.The results showed that ALK(1A4)1 + patients in 17 cases,including 8 cases of FISH(+),9 cases of FISH(-);ALK(1A4)2 + patients in 10 cases,including 7 cases FISH(+),3 cases of patients with FISH(-);ALK was not detected(1A4)3 + case.In the end,20 patients with positive FISH were detected,with a positive rate of 11%.Conclusion: 1,54 patients with ALK(1A4)lung adenocarcinoma specimens were detected for FISH antibody detection results showed that ALK(1A4)antibody sensitivity was 100%,specificity was 91.8%,and the sensitivity,specificity and the 2013 edition of the ALK NSCLC positive diagnosis expert consensus recommendations 5A4 and D5F3 anti body similar therefore,ALK(1A4)antibody can be used as ALK fusion gene positive lung cancer screening method.2,181 cases of patients with FISH results ALK rearrangement positive patients in 20 cases,the positive rate was 11%,ALK gene rearrangement occurred in the no smoking,younger(< 60 years)patients,gender,tumor cell differentiation,metastasis,clinical stage and ALK gene rearrangement was no significant difference(P > 0.05).