Screening And Validation Of Lung Adenocarcinoma Related Long Non-coding RNA Based On Profiling Microarray

Objective:Lung cancer is the highest incidence and mortality of malignant tumors in our country,80% to 90% of patients diagnosed with lung cancer in our country has been advanced,can not accept radical surgery.Although lung cancer radiotherapy,chemotherapy and molecular targeted therapy has made considerable progress,the 5 year survival rate is still less than 20%.The most important reason is that the early diagnosis of lung cancer is lack of effective means,and have the distant metastasis for lung adenocarcinoma.As other types of lung cancer,the treatment effect is not satisfactory.Therefore,it is very important to find an early diagnosis and effective treatment of lung adenocarcinoma.Long non-coding RNA is a RNA molecule with a length of more than200 nt but not able to encode proteins.Long non coding RNA polymerase II transcription RNA,but no open reading frame encoding genes with similar genetic characteristics.It is reported that the gencodev 19 human genome contains up to 13870 long chain noncoding RNA that produce more than 238981 ncrna.Long non-coding RNA is involved in the regulation of multiple diseases,and its abnormal expression and function can lead to a variety of diseases,including the occurrence of malignant tumors.Long noncoding RNA may play a role in tumor gene or tumor suppressor gene,and may be involved in the regulation of tumor proliferation,metastasis,autophagy,apoptosis and so on.It has been reported that long non-coding RNA is closely related to the progression of lung adenocarcinoma.Although some long chain non expression and function of RNA encoding in some tumors is similar,part of a long chain of non encoding has its specificity in lung cancer have specific expression profiles,finding and screening of non RNA encoding new lung adenocarcinoma,may provide a new target for the diagnosis and treatment of lung cancer.Methods: From July 1,2012 to June 31,2015,93 patients were diagnosed with lung adenocarcinoma,lung cancer tissues and the corresponding collection of the surgical resection of the adjacent normal tissues,and 3 of tissue microarray in detection of gene.Among the differentially expressed 1ncRNA,3 differentially expressed(up / down regulation of each of the 6 1ncRNA)were selected as differentially expressed1ncRNA(the expression of the difference between the two pairs of 1ncRNA was greater than 2).The construction of 1ncRNA network and mRNA expression with biological information technology at present,in order to select 1ncRNA molecules associated with lung adenocarcinoma,can learn the database through GO and KEGG function of biological information related 1ncRNA prediction,screening out key 1ncRNA is closely related with the pathogenesis of lung adenocarcinoma from.According to the screening result,methods need to be screened in the pathogenesis of lung adenocarcinoma associated 1ncRNA molecules were detected by quantitative RT-PCR,so the other 90 pairs of lung adenocarcinoma tissues and their corresponding adjacent tissues as detected by qRT-PCR,and correlated with clinicopathological parameters,such as tumor size,location,the number of lymph node metastasis,clinical stage and EGFR gene mutation status and correlation analysis.Results:In this study,we collected 3 cases with pathological analysis confirmed lung adenocarcinoma tissue samples,and select the corresponding adjacent normal tissues as control,were completed in RNA extraction,gene chip,the original data analysis,verify the chip test results,the detection of differences the results of 1ncRNA were screened and analyzed,a total of 1237 differentially expressed mRNA,up-regulated the expression of the 466,711 downregulated,594 differentially expressed 1ncRNA,134 up-regulated and 460 down regulated expression.The research results show that in lung adenocarcinoma tissues and their paired samples lung cancer tissue samples,there are significant differences on the expression of 1ncRNA.The results of differential expression of 1ncRNA were analyzed by using bioinformatics techniques and mathematical models to predict the possible biological pathways involved in differential 1ncRNA.The results showed the differentially 1ncRNA in various biological signal process of lung cancer occurrence and development,and through HLF,NF-kappaB,PAX4,COMP1,HNF-3beta and other transcription factors to regulate their target genes.Real time quantitative PCR was used to verify the differential expression of 1ncRNA,and the results were consistent with the results of microarray detection.Conclusion:1 By1ncRNA-mRNA gene chip technology was expressed in normal lung tissue and lung cancer tissue 1ncRNA and mRNA spectral data,and the scatter diagram and cluster analysis of microarray data obtained by preliminary assessment,evaluation results show the expression pattern of 1ncRNA and mRNA in the sample of lung adenocarcinoma tissues and the expression level of some specific 1ncRNA and mRNA were significantly different from normal lung tissue.2 By setting the difference expression of 1ncRNA and mRNA standard spectrum data are analyzed to determine the abnormal expression of1 ncRNA in lung adenocarcinoma tissues and mRNA spectrum data,establish the difference of 1ncRNA mRNA co expression of 1ncRNA and mRNA,through KEGG pathway analysis and GO analysis of biological information technology differential expression of functional annotation and prediction.3.It was found that LINC00152 may be the key 1ncRNA of lung adenocarcinoma,and it was confirmed by PCR that it had a significant effect on the progression of lung adenocarcinoma.Can be used as the focus of future research.