LOC285194 As Prognostic Factor Of Non-small Cell Lung Cancer And Its Mechanism

Lung cancer is one of the most tedious malignant diseasein the world.According to inadequate statistics from the World Medical Cancer Prevention and Control Agency,the overall survival rate in the last 5 years is still below 15%.Non-small cell lung cancer(NSCLC)is the major subtypes,including lung adenocarcinoma and squamous cell carcinoma.Precise Medicine and immunotherapy based on molecular marker detection can significantly reduce patients'mortality.Some small molecular markers can serve as the prognostic predictors.The study of small molecule markers has become one of the hot-spots of lung cancer research in recent years.Recent studies have shown that noncoding RNA plays an important regulatory role in the tumorgenesis and tumor development.It might be a potential biomarker for early detection and prognosis prediction in patients with NSCLC.This study has investigated the microarray data from the GEO database and re-annotated the microarray probe array using bioinformatics algorithm.The statistical data of the IncRNA expression level was obtained corresponding to the probe array by algorithm.Our analysis discover the 29 differentially expressed IncRNAs,significantly between lung adenocarcinoma and paracancerous tissues from five GSE datasets were present for the distribution of.Several different statistical analysis methods were performed.We found that the IncRNAs with maximum difference between tumor tissus and non-tumor tissue in patients with NSCLC.Furthermore we compare and analyze the GSE data sets from two categories of lung adenocarcinoma and scales carcinoma.through the intersection of different statistical analysis methods,we find some significantly different expressed lncRNAs between non-small cell lung cancer and adjacent tissues.The IncRNA LOC285194 was selected for this study.LOC285194 is also called antisense RNA LSAMP3,which is located in 3q13.31 sites,long non-coding strand RNA of approximately 2kb.LOC285194 was first reported to be an intra-tumoral suppressor in osteosarcoma.In this preliminary study,we selected 10 pairs of non-small cell lung cancer(NSCLC)samples and found that the expression of LOC285194 was significantly decreased in lung cancer tissues relative to paracarcinoma tissues.So,LOC285194 was lowly expressed in non-small cell lung cancer tissues(P<0.05).Next,we investigate the expression of LOC285194 in NSCLC cell lines(A549,NCI-H1299,NCI-H460,PC-9,Calu-3)and normal bronchial epithelial cell lines(16HBE).Also we found non-small cell lung cancer cell lines with low expression level of LOC285194.It is sure that the expression of LOC285194 was significantly different in non-small cell lung cancer cell lines(P*<0.001).We identified the functional silencing sequence of siLOC285194 by siRNA technique and constructed the LOC285194-shRNA-A549 cell line(Lentivirus-infected "hairpin RNA" interferes with knockdown of LOC285194-A549 cell line)following functional knockdown sequences,and studied the cellular mechanisms.Functional and mouse in vitro experimental models.We detected LOC285194 expression levels in lung cancer cell line LOC285194-shRNA-A549 treated with gene knockdown technology and found that they were significantly downregulated compared to the transfected empty plasmid control group.Cell function tests showed that the capacity of proliferation and migration was significantly enhanced in knockdown cell line LOC285194-shRNA-A549.It was confirmed that down-regulation of LOC285194 expression increased tumor cell proliferation and migration.Nude mouse tumorigenicity experiment was also performed to test the function of LOC285194.We found that,tumor with LOC285194-shRNA presented higher growth rate than those with blank control group.(P<0.05).It was suggested that the lung cancer cell line treated with the gene silencing/knockdown technique for LOC285194,with the down-regulation of LOC285194 and the ability to control the proliferation or growth of the tumor.It was found that LOC285194 was a tumor suppressor in animals,which would inhibit metastasis in vivo.In order to further understand the prognostic significance of LOC285194 in the actual disease of non-small cell lung cancer.In this study,tissue samples from 56 non-small cell lung cancer cases and matched paired paraneoplastic lung tissues were detected.All specimens were detected by qPCR to detect the expression level of RNA and found:in all 56 pairs of non-small cell lung cancer tissues.Relative to adjacent tissues,the low expression cases were 100%.Based on the median expression level of LOC285194,56 pairs of samples were divided into low expression group and high expression group.Kaplan-Meier analysis,Cox analysis,and X test was performed.The results showed that in the real clinical disease case,the PFS and OS were better in the high expression group than in the low expression group by the survival analysis and the relationship with the clinicopathological features.There was a significant correlation between the expression level of LOC285194 and the T stage of the tumor,but there was no correlation with "age","gender","clinical stage",and "type of tissue".The survival rate of patients with low expression of LOC285194 in patients with non-small cell lung cancer was significantly lower than that in the high expression group.Low expression of LOC285194 was one of the independent indicators of poor prognosis of NSCLC.This indicates that LOC285194 may be associated with lung cancer cell function and prognosis.It is not yet clear that LOC285194 may cause any unique characteristics and molecular mechanisms associated with human cancer.More in-depth research meirt.This article through the previous period of the GEO database chip analysis combined with literature analysis found that:LOC285194 and tumor suppressor gene p53 pathway,we have LOC285194 as a tumor cell function mechanism research found that by LOC285194 RNA Pull-down analysis shows that the captured lncRNA-Protein Complex-has a band with a molecular weight of P53.WB experiments detected the presence of P53 protein.Further detection of LOC285194 by qPCR after RNA immunoprecipitation captures the lncRNA-protein complexby antibody against P53.The P53 protein binds to the 3'-end domain of LOC285194 and studies confirm that LncRNA-LOC285194 and P53 are present in non-small cell cells.Lung cancer in combination with each other,suggesting that LOC285194 is associated with P53 pathway in non-small cell lung cancer,its high expression can activate the P53-related pathway,and exercise its biological characteristics of inhibiting the proliferation and migration of lung cancer cells.LOC285194 is a tumor suppressor unit and an independent prognostic factor in non-small cell lung cancer.In translational medicine,how to develop RNA-targeted drugs and find a translational medical method for overexpression of LOC285194 should be very important.