Effect of circTHBS1 on Malignant Biological Behaviors of Breast Cancer Cells by Regulating miR-135a-5p/RAB1A Axis
This study aims to investigate the effect of circTHBS1(circular RNA thrombospondin-1)on the ma-lignant biological behaviors of BC(breast cancer)cells by regulating the miR-135a-5p(microRNA-135a-5p)/RAB1A(Rab family protein 1A)axis.qRT-PCR(real-time fluorescence polymerase chain reaction)was used to detect the expression of circTHBS1,miR-135a-5p,and RAB1A mRNA in BC cell lines,and the optimal cell line was screened for subsequent experiments.MCF-7 cells were grouped into Control group,sh-NC group,sh-THBS1 group,sh-THBS1+inhibitor NC group,and sh-THBS1+miR-135a-5p inhibitor group.qRT-PCR was used to detect transfection efficiency.Dual lucifer-ase reporter gene method was used to confirm the interaction between circTHBS1 and miR-135a-5p,miR-135a-5p and RAB1A.Western blot assay was used to detect the expression of RAB1A in MCF-7 cells.Plate cloning method was used to detect the proliferation of MCF-7 cells.Flow cytometry was used to detect the apoptosis of MCF-7 cells.Scratch as-say was used to detect the migration of MCF-7 cells.Transwell method was used to detect the invasion of MCF-7 cells.The nude mouse transplantation tumor experiment was used to detect the effect of circTHBS1 on the growth of BC cells in tumor tissue.The dual luciferase reporter gene assay found a targeted relationship between circTHBS1 and miR-135a-5p,RAB1A and miR-135a-5p.Compared with normal breast epithelial cells MCF-10A,the expression of circTHBS1 and RAB1A obviously increased in BC cell lines,while the expression of miR-135a-5p obviously decreased(P＜0.05).Knock-ing down the expression of circTHBS1 was able to inhibit the proliferation,migration,and invasion of MCF-7 cells,and induce MCF-7 cell apoptosis(P＜0.05).Inhibiting the expression of miR-135a-5p was able to reverse the inhibitory effect of knocking down circTHBS1 expression on the proliferation,migration,and invasion of MCF-7 cells,and inhibit MCF-7 cell apoptosis(P＜0.05).Through nude mouse transplantation experiments,it was found that knocking down the expression of circTHBS1 resulted in a decrease in the mass and volume of BC tumor tissue in nude mice,and a decrease in RAB1A expression and an increase in miR-135a-5p expression(P＜0.05).circTHBS1 is highly expressed in BC cells.Knocking down circTHBS1 can inhibit the malignant biological behaviors of BC cells by regulating the miR-135a-5p/RAB1A axis.
circular RNA thrombospondin-1microRNA-135a-5pRab family protein 1Abreast cancerproliferationapoptosismigrationinvasion
万方数据
维普
