LncRNA DICER1-AS1 Targeting Inhibits DICER1 Promotes Lung Cancer Proliferation,Invasion And Metastasis

Objective:Lung cancer is a kind of malignant tumor derived from the malignant changes of bronchial epithelial cells.It has become the third most common malignant tumor in the world,and we are looking for a molecular target for early diagnosis and treatment.With the development of high-throughput sequencing technology,IncRNA DICER1-AS1 was found to be abnormally expressed in tumor tissues of lung cancer patients.The aim of this study was to reveal the influence of abnormal expression of LncRNA DICER1-AS1 in lung cancer on the diagnosis and treatment of lung cancer patients,and to explore the molecular mechanism of proliferation,invasion and metastasis,and provide reference for clinical diagnosis and treatment of lung cancer.Methods:1.The RNA was extracted from the cancer tissues and adjacent tissues of 66 lung cancer patients with pathological examination confirmed by the Third Affiliated Hospital of Kunming Medical University.The expression of IncRNA DICER1-AS1 mRNA in two tissues was detected by QPCR,and we analysis it combining with clinical data;mRNA expression of LncRNA DICER1-AS1 in three cells was detected by QPCR after RNA was extracted from lung cancer cell line A549,H1299 and human embryonic kidney diploid cell KMB17.2.Lentivirus infected lung cancer cell line A549 and H1299 to express shRNA knocking down IncRNA DICER1-AS1.That we use CCK8,transwell,scratch test and colony formation experiments to observe whether knocking down DICER1-AS1 would affect the related organisms of these tumor cells Phenotype;3.IncRNA DICER1-AS1 as DICER1 antisense transcript may target DICER1,we also use QPCR to detect the expression of DICER1 mRNA in the above 66 cases of lung cancer and adjacent tissues,we use RT-PCR to detect mRNA of DICER1 and western bolt to detected protein of DICER1 expression after knocking down lncRNA DICER1-AS1.Results:1.QPCR results showed that the mRNA expression of IncRNA DICER1-AS1 in lung cancer tissues 0.94(0.48,1.70)was higher than that in adjacent tissues 0.65(0.45,0.86),p<0.05,A549 mRNA expression(23±2.7),H1299 mRNA expression(42.3±0.48)were higher than that of human diploid normal cells(KMB17)mRNA(5.84±1.07),p<0.001.2.Clinical sample data showed that DICER1-AS1 in female lung cancer tissue’s expression level was higher than that of males(p<0.05),but there was no significant difference in TNM stages,ages and tumor sizes(p>0.05).3.The results of knockdown of DICER1-AS1 in lung cancer cells showed that the CCK8 was detected at 450 nm,and the absorbance of the DICER1-AS1 group was lower than that of the control group(p<0.05).The result of transwell was that the experimental group with the knockdown of DICER1-AS1 the invasive ability was weaker compared with the control group,(p<0.05);the cell clone formation experiment showed that the experimental group of the DICER1-AS1 knockdown decreased the cell colonies compared with the control group(p<0.05);the scratch test showed that the experimental group showed a comparison with the control group.The cell migration ability was weaker.4.The expression of DICER1 in lung cancer tissues7.5(5.29,10.23)was lower than that in adjacent tissues 10.11(5.78,15.12),p<0.05.RT-PCR and western bolt results showed that DICER1-AS1 was knocked down.The level of DICER1 protein was up-regulated after knocked down DICER1-AS1(p<0.001),and there was no significant change in mRNA level(p>0.05).Conclusion:1.LncRNA DICER1-AS1 is highly expressed in clinical lung cancer tissues and may play an important role in cancer-promoting genes in lung cancer,and may be related to gender.2.lncRNA DICER1-AS1 can promote the invasion,proliferation,Metastasis of lung cancer cells;3.IncRNA DICER1-AS1 may target the inhibition of DICER1 on the invasion,proliferation and metastasis of lung cancer cells;4.IncRNA DICER1-AS1 promotes the occurrence and development of lung cancer,and is expected to become a potential target for clinical diagnosis and treatment.