| ObjectiveDetection of venous blood tumor markers,sputum cytology,bronchoalveolar lavage fluid(BALF)cytology and bronchoalveolar lavage fluid of short stature homobox 2(SHOX2)and Ras-association domain family 1A(RASSF1A)genes the sensitivity of various indicators such as methylation in peripheral lung cancer and benign lung lesions,analysis of the diagnostic efficacy of single and multiple indicators combined detection for peripheral lung cancer and benign lung diseases,evaluation of short stature homobox 2(SHOX2)and Ras-association domain family 1A(RASSF1A)dual gene methylation combined detection in the clinical application value of peripheral lung cancer diagnosis.MethodsThe 102 patients with peripheral lung cancer who were treated in the First Affiliated Hospital of Jinzhou Medical University and Chifeng University Affiliated Hospital from July 2019 to December 2020 were selected as the observation group;The 66 patients with benign lung lesions in the two hospitals during the same period were selected as the control group.After all subjects are enrolled in the group,collect general demographic data and compare the differences in general data between the two groups;collect bronchoalveolar lavage fluid(BALF)of all patients,using real time polymerase chain reaction(RT-PCR)and Sanger sequencing at the same time method two methods for SHOX2 and RASSFlA gene methylation detection,statistical analysis of the consistency of the test results of the two detection methods,comparison of the positive rate of double gene methylation detection in the two groups of patients;collection of venous blood of the observation group and the control group,sputum and bronchoalveolar lavage fluid(BALF),compare venous carcino embryonic antigen(CEA),cytokeratin 19 fragment(CYFRA21-1),neuron-specificenolase(NSE),sputum cytology,bronchoalveolar perfusion the sensitivity and specificity of BALF cytology and its SHOX2 and RASSF1A gene methylation and other indicators,and then use receiver operating characteristic curve(ROC)to explore the combined detection of single or multiple indicators.The diagnostic efficacy and clinical application value of type lung cancer,and through multivariate regression analysis of lung cancer-related risk factors,a lung cancer prediction model was established.Results1.There was no statistically significant difference between the observation group and the control group in terms of gender,age,marital status,and place of residence(P>0.05);the observation group and the control group had statistical differences in smoking history and tumor family history significance(P<0.05).2.For all subjects,RT-PCR and Sanger sequencing were used to detect the double gene methylation of SHOX2 and RASSF1A in bronchoalveolar lavage fluid(BALF).Among them,the SHOX2 methylation test,Kappa=0.868(P<0.001);RASSF1A methylation detection,Kappa=0.819(P<0.001).3.The positive rate of SHOX2 observation group was 74.5%,the positive rate of control group was 7.6%;the positive rate of RASSF1A observation group was 59.8%,and the positive rate of control group was 3.0%.The observation group and the control group had differences in SHOX2 and RASSF1A.Statistical significance(P<0.001);SHOX2 and RASSF1A methylation combined detection,the positive rate of observation group was 85.3%.,The control group was 10.6%,and the difference between the observation group and the control group in the combined detection of SHOX2 and RASSF1A methylation was statistically significant(P<0.001).4.The observation group was positive in venous blood CEA,CYFRA21-1,NSE,sputum cytology,bronchoalveolar lavage fluid cytology,SHOX2 gene methylation,RASSF1A gene methylation,SHOX2,RASSF1A gene methylation combined detection the rates were 38.2%,30.4%,14.7%,19.6%,42.2%,74.5%,59.8%,85.3%,and the control group were 3.0%,3.0%,1.5%,1.5%,3.0%,7.6%,3.0 %,10.6%,the difference between the observation group and the control group in the above detection methods is statistically significant(P <0.001).5.Among different pathological types,the positive rates of SHOX2 gene methylation in adenocarcinoma,squamous cell carcinoma,small cell carcinoma,and other types of cancer are 75.0%,76.5%,77.8%,and 50.0% respectively;the positive rate of RASSF1A gene methylation is respectively 59.7%,58.8%,66.7%,50.0%;SHOX2,RASSF1A gene methylation combined detection positive rates were 84.7%,88.2%,88.9%,75.0%,the positive rate was significantly higher than any single gene a baseline test results;patients with stage I-II and patients with stage III-IV,in venous blood CEA,bronchoalveolar lavage fluid cytology,SHOX2 gene methylation,RASSF1A gene methylation,combined detection of SHOX2 and RASSF1A genes the difference in positive rate was statistically significant(P<0.05).6.Using ROC curve for comparative analysis,venous blood CEA,bronchoalveolar lavage fluid cytology,SHOX2 gene methylation,RASSF1A gene methylation,combined detection of SHOX2 and RASSF1A genes,and bronchoalveolar lavage fluid cytology,SHOX2 combined detection of gene methylation and RASSF1A gene methylation,the AUC values were 0.676,0.696,0.835,0.784,0.873,0.893.7.Through multivariate logistic regression analysis,bronchoalveolar lavage fluid cytology,SHOX2 gene methylation,RASSF1A gene methylation,tumor family history,smoking history entered the regression equation(P<0.05).Conclusions1.The detection of SHOX2 and RASSF1A gene methylation in bronchoalveolar lavage fluid is more sensitive than venous blood tumor markers,sputum and bronchoalveolar lavage fluid cytology in the diagnosis of peripheral lung cancer;2.The combined detection of SHOX2 and RASSF1A double gene methylation in bronchoalveolar lavage fluid can significantly increase the positive diagnosis rate of peripheral lung cancer.It is an effective supplementary tool for the pathological diagnosis of peripheral lung cancer and is expected to become a new marker of peripheral lung cancer.3.Bronchoalveolar lavage fluid cytology,SHOX2 gene methylation,RASSF1A gene methylation,tumor family history,smoking history can predict whether an individual has lung cancer to a certain extent. |
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