ABSTRACT
This research aimed to explore the molecular mechanism of microRNA (miR)-106b in cell apoptosis of atherosclerosis (AS). Human aortic endothelial cells (HAECs) were divided into control group, oxidized-low-density lipoproteins (ox-LDL) group, miR-106b NC+ox-LDL group, miR-106b mimics+ox-LDL group, miR-106b mimics+PTEN+ox-LDL group, and miR-106b mimics+empty+ox-LDL group. Real-time fluorescence quantitative polymerase chain reaction, cholecystokinin, TdT-mediated biotinylated nick end-labeling assay, luciferase reporter gene assay, and flow cytometry analysis were performed to determine the morphology, proliferation, and apoptosis in HSECs. Moreover, the levels of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), Bcl-2, p-P13K, and p-AKT in HAECs were detected by western blot. MiR-106b was down-regulated in ox-LDL-induced HAECs. PTEN was the target gene of miR-106b-5p. Overexpression of PTEN inhibited the anti-apoptotic effect of miR-106b. Compared with the control group, the proportion and number of HAECs apoptosis and Bax, caspase-3, and caspase-9 expression in ox-LDL and miR-106b mimics+PTEN+ox-LDL groups were significantly increased (all P<0.05). Moreover, the activity of HAECs and Bcl-2 were decreased significantly (all P<0.05). Overexpression of miR-106b in ox-LDL-induced AS inhibited endothelial cell apoptosis. Furthermore, miR-106b might activate the PI3K/AKT pathway by down-regulating the expression of PTEN in ox-LDL-induced HAECs.
Subject(s)
Humans , Apoptosis , MicroRNAs/genetics , Endothelial Cells/metabolism , Atherosclerosis/metabolism , Lipoproteins, LDL/genetics , Signal Transduction , Up-Regulation , Cell Proliferation , Real-Time Polymerase Chain Reaction , Fluorescence , Lipoproteins, LDL/metabolismABSTRACT
Aim To investigate the effect of cinobufagin on the migration and invasion of esophageal cancer Kyse-520 cells, and to explore the underlying molecular mechanism. Methods The rates of inhibition after treated with different concentrations of cinobufagin 12, 24, 48 h were detected by CCK-8 method, and the changes of cell migration and invasion were observed with wound healing and transwell assay. The mRNA expressions of FAK, Akt, PTEN, VEGF-A, MMP-2 and MMP-9 were detected by quantitative real-time polymerase chain reaction ( RT-qPCR ). The protein expressions of FAK, p-FAK ( Tyr397 ), Akt, p-Akt (Sei473 ), VEGF-A, PTEN, MMP-2 and MMP-9 were measured by Western blot. Results The results of CCK-8 showed that cinobufagin could inhibit the proliferation of Kyse-520 cells in a time- and concentration-dependent manner, and cinobufagin significantly inhibited the cell invasion and migration. Meanwhile, data from RT-qPCR and Western blot suggested that cinobufagin had no significant effect on mRNA and total protein of FAK and Akt, but it reduced the expression of p-FAK(Tyr397), p-Akt(Ser473), VEGF- A, MMP-2 and MMP-9, and increased the PTEN expression. Conclusions Cinobufagin significantly inhibits the invasion and migration of esophageal cancer Kyse-520 cells through inducing PTEN expression and down-regulating FAK/PI3K/AKT pathway.