| BackgroundGlobally,lung cancer,as the second most morbidity and highest mortality malignant tumor,seriously endangers human life and health,and has always been the focus of research.The pathogenesis of lung cancer is relatively complicated.At present,it is believed that the occurrence of lung cancer is closely related to factors such as genetic factors,environmental pollution exposure,food,smoking,and lifestyle.In terms of histopathology,lung cancer is divided into small cell lung cancer(SCLC)and non-small cell lung cancer(NSCLC).Among them,non-small cell lung cancer accounts for about 85%.Among them,lung adenocarcinoma(Lung Adenocarcinoma,LUAD)is the most common histological subtype of lung cancer.At present,the treatment of lung adenocarcinoma is mainly surgery,radiotherapy,chemotherapy and targeted therapy.Surgical treatment is the early and mid-stage lung glands.However,some patients are already at an advanced stage when they are diagnosed with lung adenocarcinoma,and radiotherapy and chemotherapy will produce gastrointestinal reactions,bone marrow suppression and other adverse reactions.Therefore,the survival time of lung adenocarcinoma is still very low.Therefore,finding new therapeutic targets is of great significance for the prognosis of lung adenocarcinoma.LncRNA is a type of RNA that cannot be encoded and is greater than 200 nucleotides in length.In recent years,lncRNA has been confirmed to play a regulatory role in gene modification,chromosome silencing,and genome imprinting.LncRNA has been a research hotspot in recent years.It has been found to be abnormally expressed in a variety of tumor tissues,which affects the biological effects of cells.In our research,we also found that lncRNA DDX11-AS1 plays an important role in lung adenocarcinoma,and is involved in regulating the proliferation and metastasis of lung adenocarcinoma.In this study,we are divided into two parts.The first part detects the expression level and clinical significance of lncRNA DDX11-AS1 in lung adenocarcinoma,and the second part explores the molecular mechanism of lncRNA DDX11-AS1 regulating the proliferation and metastasis of lung adenocarcinoma cells.Part Ⅰ The expression and clinical significance of LncRNA DDX11-AS1 in lung adenocarcinoma tissues and cellsObjectTo study the expression of lncRNA DDX11-AS1 in lung adenocarcinoma,and further analyze its clinical significance.Method1 From the tumor gene databases of TCGA(https://portal.gdc.cancer.gov/)and GEO(https://www.ncbi.nlm.nih.gov/geo),screen for differentially expressed genes.2 Using qRT-PCR technology to detect the expression of lncRNA DDX11-AS1 in lung adenocarcinoma cells A549 and H1299.3 Using the tumor clinical data from the TCGA database,analyze the relationship between the expression level of lncRNA DDX11-AS1 and the clinical characteristics and prognosis of lung adenocarcinoma.Result1 Compared with adjacent tissues and cells,the expression level of lncRNA DDX11AS1 in lung adenocarcinoma tissues and cells was up-regulated,and the difference was statistically significant(P<0.05).2 The expression level of LncRNA DDX11-AS1 is related to the age of lung adenocarcinoma(P<0.05),clinical stage(P=0.006),and lymph node metastasis(P=0.04).3 The 5-year survival rate of lung adenocarcinoma with high expression of lncRNA DDX11-AS1 is lower than that of the low expression group(P<0.05),and lncRNA DDX11-AS1 is an independent prognostic factor.Part Ⅱ Explore the molecular mechanism of LncRNA DDX11-AS1 regulating the proliferation and migration of lung adenocarcinoma cellsObjectTo explore the effect of lncRNA DDX11-AS1 on the proliferation and migration of lung adenocarcinoma cells,and to explore the potential molecular mechanism of lncRNA DDX11-AS1 in the occurrence and development of lung adenocarcinoma.Method1 Using molecular biology technology to design si-lncRNA DDX11-AS1 to transfect lung adenocarcinoma cells A549 and H1299,and use qRT-PCR to detect the knockdown effect of lncRNA DDX11-AS 1.2 CCK-8 and clone formation experiments were used to detect the effect of lncRNA DDX 11-AS 1 on the proliferation of lung adenocarcinoma cells A549 and H1299.3 The scratch test and Transwell test were used to detect the change of lncRNA DDX11AS1 on the migration ability of lung adenocarcinoma cells A549 and H1299.4 Predict miRNAs that may be specifically adsorbed to lncRNA DDX11-AS1 in the Starbase database,and use qRT-PCR to detect miRNA expressionResult1 The qRT-PCR test results showed that compared with the si-NC group,the expression levels of lncRNA DDX11-AS1 in the si-lncRNA DDX11-AS1#1 and#2 groups were significantly down-regulated,and the difference was statistically significant(P<0.05).2 CCK-8 and clone formation experiments showed that compared with the si-NC group,the proliferation ability of lung adenocarcinoma cells A549 and H1299 in the silncRNADDX11-AS1＃1 and#2 groups was significantly weakened,and the difference was statistically significant(P<0.05).3 Scratch test and Transwell test showed that compared with the si-NC group,the migration ability of lung adenocarcinoma cells A549 and H1299 in the si-lncRNA DDX11-AS1#1 and#2 groups was significantly weakened,and the difference was statistically significant(P<0.05).4 The qRT-PCR test results showed that compared with the si-NC group,the expression of miR-766-5p in lung adenocarcinoma cells A549 and H1299 in the si-lncRNA DDX11-AS1#1 and#2 groups was significantly up-regulated,and the difference was statistically significant(P<0.05).Conclusion1 LncRNA DDX11-AS1 is highly expressed in lung adenocarcinoma tissues and cells,and the expression level is related to the age,clinical stage,and lymph node metastasis of patients with lung adenocarcinoma.In addition,patients with lung adenocarcinoma with high expression of lncRNA DDX11-AS1 have a poor survival prognosis,and 1ncRNADDX11-AS1 is an independent prognostic factor.2 Knockdown of lncRNA DDX11-AS1 in vitro inhibits the proliferation and migration of lung adenocarcinoma cells A549 and H1299.3 LncRNA DDX11-AS1 can regulate the expression level of miR-766-5p. |
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