Study On The Mechanism Of TRF-Val-CAC And Its Binding Protein RAC1 In Lung Adenocarcinoma Cells

Objective(s):More and more studies have found that it plays important roles in many aspects.In order to further explore the mechanism of tRF-Val-CAC regulation in lung adenocarcinoma,the binding protein of tRF-Val-CAC was screened out,the molecular silencing lentiviral vector was designed and synthesized and co-transfected with the previously designed tRF-Val-CAC inhibitor to verify the regulatory relationship between tRF-Val-CAC and its binding protein,and to verify the synergistic regulation of tumor cell proliferation,invasion and migration.Methods:First,tRF-Val-CAC binding proteins were screened byRNA pull-down,liquid chromatography-mass spectrometry,and bioinformatics analysis.Its expression was then verified by qRT-PCR and Western Blot.Then,the RAC1 silencing vector was synthesized and a stable cell line was constructed,which was co-transfected with tRF-Val-CAC inhibitor to verify the regulatory relationship between tRF,silencing binding proteins and co-silencing the two.Through the detection of cell proliferation by CCK-8,Transwell invasion and cell scratch experiment,cell invasion and migration were verified,to verify the regulatory effect of the two on the phenotypic function of tumor cells.Results:The tRF-Val-CAC binding protein RAC1,which is associated with invasion and migration,was screened for high expression in lung adenocarcinoma cells.The construction of RAC1 knockdown stable transfected cells inhibits the expression of RAC1RNA and inhibits RAC1 phosphorylation without affecting protein expression.Experimental data from qRT-PCR and Western Blot showed that the expression of tRF-Val-CAC increased when the expression of RAC1 was inhibited,and the expression of tRF-Val-CAC did not affect the expression of RAC1,and the expression of tRF-Val-CAC may be related to the phosphorylation level of RAC1.CCK8 cell proliferation experiments showed that inhibition of RAC1 expression inhibited the proliferation of cells.Transwell invasion experiments and scratch experiments showed that the low expression of tRF-Val-CAC and the knockdown of RAC1 inhibited cell invasion and migration,with the former having a higher rate of cell invasion inhibition and the latter having a higher rate of cell migration inhibition.Conclusion(s):When the phosphorylation of RAC1 decreases,the expression of tRF-Val-CAC increases instead of RAC1,which further promotes the invasion and migration of tumor cells,thereby promoting tumorigenesis and development.