Screening And Verification Of Susceptibility Genes For Lung Adenocarcinoma And Exploration Of Its Underlying Mechanism

Objective:To screen the specific susceptibility genes for lung adenocarcinoma and explored its biological functions based on literature research and data analysis.Methods:This study,human lung bronchial epithelial cells Beas-2b,Xuanwei lung cancer cells XWLC-05,human non-small cell lung cancer cells NCI-H1299,SPC-A1,A549,and the clinical tissues and serum of lung cancer patients from Yunnan Cancer Hospital were used as models:1.The qRT-PCR technique was used to verify the reverse transcription of candidate genes on lung adenocarcinoma cells,and the lung adenocarcinoma cDNA microarray chips with clinical follow-up were used to perform tissue reverse transcription verification on the candidate genes to further screen the differential genes.2.Western Blot was preformed to verify the protein expression of differential genes on lung adenocarcinoma cells.3.The qRT-PCR technique and immunohistochemical staining methods were used to analyze the reverse transcription of differential genes and protein exprssion of the postoperative tissues of adenocarcinoma patients collected from Yunnan Cancer Hospital.4.Combining RNA purification technology and qRT-PCR to analyze the expression of differential genes in the serum of clinical lung adenocarcinoma patients.5.Analyze the correlation between differential genes and lung adenocarcinoma and possible potential regulatory mechanisms through bioinformatics..Results:1.Compared with the Beas-2b cell line,LAD1,COL10A1 and IGSF-9 were different in the reverse transcription level of the other four lung cancer cells,and they were statistically significant(P<0.05).LAD1,FAM111B,and COL10A1 were significantly overexpressed in XWLC-05 lung cancer cells compared with Beas-2b cell line(P<0.05).LAD1,FAM111B,IGSF9,and COL10A1 all have significant gene overexpression differences in lung adenocarcinoma tissues,and FAM111B showed 11.4 times the relative expression level compared to the negative group,and the difference was statistically significant(P<0.05).2.In terms of protein expression,FAM111B was almost not expressed in the negative control Beas-2b cells,but showed high expression in the four lung cancer cell lines,and it was more significant in XWLC-05 lung cancer cells(P<0.05).3.FAM111B showed an 8.5-fold difference in reverse transcription expression in lung cancer tissues compared to the negative control in 51 selected lung cancer patients(P<0.05),who were from the lung cancer patients of Yunnan Provincial Tumor Hospital from December 2018 to February 2020,Where:LUAD,n=25;SCLC,n=26.The expression of FAM111B in lung squamous cell carcinoma tissue was higher than that in lung adenocarcinoma(P<0.05).The results of immunohistochemistry showed that the IRS score of FAM111B in lung adenocarcinoma tissues(Ca Group)was significantly higher than that of the control group(Para-Ca group)(P<0.05).4.After RNA purification of the paired sera,we found the presence of free RNA in the sera of lung adenocarcinoma patients and non-cancer patients.We did not find the reverse transcription expression of FAM111B in the serum of lung adenocarcinoma patients and non-cancer patients by using P53 as an external reference control.5.Combining the above experimental results,clinical lung adenocarcinoma patients’medical records and bioinformatics databases to conduct bioinformatics analysis of the relationship between FAM111B and lung adenocarcinoma,we found that the expression of lung adenocarcinoma patients in stage Ⅱ and Ⅲ is higher than that in stage Ⅰ and Ⅳ,but there is no significant correlation.Analysis of follow-up cases showed that the survival time and survival rate of patients with lung adenocarcinoma with high FAM111B expression decreased significantly(HR=1.49(1.26-1.75),log-rank P=2×10-6).Conclusions:1.The reverse transcription and protein levels of FAM111B is significantly higher in lung cancer cells compared to the normal lung epithelial cells,and more prominent in lung adenocarcinoma cells.2.In lung adenocarcinoma tissues,the reverse transcription and protein levels of FAM111B were significantly up-regulated,but the corresponding free RAN was not detected in the serum.3.According to bio-informatics analysis,FAM111B is a postoperative survival risk factor for lung adenocarcinoma,and it is expected to be used as a clinical postoperative survival index for lung adenocarcinoma combined with the above results.