Molecular Study Of The Expression Of MiR-194-5p In Lung Adenocarcinoma Tissues And Its Regulation Of Cell Proliferation And Invasion

ObjectiveThe incidence and mortality of lung cancer has been at the forefront in the world,and it is the largest cancer in China.It has great clinical significance for early treatment of lung cancer patients to improve the overall survival benefit of patients.However,due to its complex biological behavior and pathogenesis,Lack of specific molecular markers for early diagnosis,Most patients have progressed or transferred at the time of first diagnosis,Therefore,the deep investigation of the pathogenesis of lung cancer,Finding relevant diagnosis and treatment targets is important,In this study,mainly by comparing the expression of mi R-194-5p in lung cancer tissues and adjacent normal tissues,To analyze the clinical significance of the expression of mi R-194-5p for lung cancer,And then study the regulation of the biological functions of lung cancer cells from the level of cells and animals,respectively,Further improve the molecular mechanism of lung cancer,To provide a new reference and direction for finding new diagnosis and treatment targets of lung cancer.Methods(1)The expression of mi R-194-5p in lung cancer tissues and normal tissues was detected by quantitative fluorescence PCR technology,and the correlation between mi R-194-5p expression and clinicopathology was analyzed.(2)The expression of mi R-194-5p in common lung cancer tumor cell lines(A549 cells,PC-9 cells,H1299 cells and H1795)was detected by fluorescence quantitative PCR,and PC-9 cells with low expression of mi R-194-5p were selected for functional study of this gene.(3)Simulics of mi R-194-5p and blank controls were then transfected into PC-9 cells to construct a mi R-194-5p overexpressing cell line.(4)The effect of mi R-194-5p on the proliferation ability of lung cancer cells was tested by CCK 8,the effect of mi R-194-5p on the clone formation ability of lung cancer cells,and the ability of mi R-194-5p by Transwell and scratch-assay,and the effect of mi R-194-5p on protein expression related to invasion and migration of lung cancer cells by Western blot.(5)Bioinformatics was used to predict the downstream target genes of mi R-194-5p,and the targeting of mi R luciferase-194-5p was verified by using dual luciferase reporter gene,fluorescence quantitative PCR,Western blot,clone formation experiment,Transwell experiment and in vivo nude mice.Results(1)The relative expression level of mi R-194-5p in lung cancer tissues was 1.72 ± 0.64,Significantly lower than in the adjacent tissue(3.84 ± 0.95),Statistically significant difference(P <0.05),In the 30 lung cancer samples,With significant differences in adjacent and lung cancer tissues,In 19(63.3%)patients,the lowly expressed mi R-194-5p,Mi R-194-5p in 11(36.7%)patients was highly expressed,Expression of mi R-194-5p with the sex and age of lung cancer patients,The degree of differentiation and smoking history were not correlated,However,a significant association with TNM stage(P = 0.0024)and lymph node-related metastasis(P = 0.0086),Low expression of mi R-194-5p.(2)The quantitative PCR showed that the transfection of mi R-194-5p could significantly promote the expression level of mi R-194-5p in PC-9 cells compared with the blank control.(3)The results of CCK-8 showed that the transfection of mi R-194-5p could significantly inhibit the proliferation capacity of PC-9 cells compared with the blank control.(4)The results of the clone-formation assay showed that the transfection of mi R-194-5p could significantly inhibit the clone-formation ability of PC-9 cells.(5)The results of Transwell and scratch experiments showed that the transfection of mi R-194-5p could significantly inhibit the invasion and migration ability of PC-9 cells compared with the blank control.(6)Western blot results showed that transfection of mi R-194-5p could significantly promote E-cadherin expression in PC-9 cells compared with the blank control.(7)Bioinformatics and dual-luciferase reporter gene prediction found that mi R-194-5p could target binding and regulate the Ras-related protein Rad-38(RAB 38).(8)The Western blot assay showed that the transfection of mi R-194-5p could significantly inhibit the expression of RAB38 protein level in PC-9 cells compared with the blank control.(9)Clone formation experiments,Transwell experiments and in vivo nude mice experiments found that mi R-194-5p inhibited the proliferation and invasion ability of lung cancer cells by negatively regulating the expression of RAB 38.ConclusionAfter finding that the relative expression level of mi R-194-5p was significantly lower in lung adenocarcinoma tissues than in normal tissues,Of mi R-194-5p expression and the sex and age of patients with lung adenocarcinoma,The degree of differentiation and smoking history were not correlated,But showed a significant association with TNM stage and lymph node-related metastasis,The vast majority of late stage and patients with lymph node metastasis showed low expression of mi R-194-5p,Upregulation of mi R-194-5p expression significantly inhibited the proliferation,migration and invasion capacity of lung adenocarcinoma cells,The mechanism may be achieved through the negative regulation of RAB 38 protein expression,Of mi R-194-5p can act as a tumor suppressor gene,As a potential biomarker for the diagnosis and treatment of lung adenocarcinoma.