RESUMO
【Objective】 To investigate the effects of miR-126-3p targeting chemokine receptor 1 (CCR1) in exosomes derived from bone marrow mesenchymal stem cells (BMSC) on the proliferation, migration, and invasion of lung cancer cells. 【Methods】 BMSC cells were cultured; exosomes were extracted and identified by the exosomal marker proteins CD63 and TSG101. After exosome culture of A549 cells for different durations (0, 24, 48, and 72 h), cell survival rate was detected by CCK-8, mRNA levels of miR-126-3p and CCR1 were detected by qRT-PCR, and cell migration and invasion abilities were detected by Transwell assay. The relative expressions of CCR1, epithelial cadherin (E-cad), neural cadherin (N-cadherin), and Vimentin were detected by Western blotting. 【Results】 Exosomes had round or oval cup-shaped structures with bright edges and dark middle, with a particle size distribution of about 152 nm, expressing CD63 and TSG101 proteins. The expression of miR-126-3p in exosomes was higher than that in A549 cells. The expression of miR-126-3p was low in A549 cells and that of CCR1 mRNA was high. However, after co-culture with exosomes, the expression of miR-126-3p in A549 cells was increased, while the expression of CCR1 was decreased. A549 cells were cocultured with exosomes for 0, 24, 48, and 72 h. The survival rate, migration and invasion abilities, CCR1 gene and protein expression levels, and N-cad and Vimentin protein expression levels of A549 cells decreased gradually with the extension of culture time. The level of miR-126-3p and the expression of E-cad protein increased gradually with the extension of culture time. 【Conclusion】 The co-culture of exosomes derived from bone marrow mesenchymal stem cells with A549 cells can increase the expression level of miR-126-3p, and miR-126-3p can reduce the proliferation, migration, and invasion of A549 cells by targeting the inhibition of CCR1 expression.
RESUMO
Objective:To investigate the expression of miR-126-3p in thyroid cancer and the biological function.Methods:The expression of miR-126-3p in thyroid carcinoma,adjacent tissues and three types of thyroid cancer cells(TPC-1,FTC-133,8505C) were detected by RT-PCR;thyroid cancer cells were divided into analogue group(mimic) and control group(NC),which were respectively with the transfection of miR-126-3p mimic and negative control plasmid.The proliferation and apoptosis in two groups were respectively detected by Brdu-ELISA and flow cytometry.The migration and invasion were detected by Transwell Chambers method.Results:The expression of miR-126-3p in thyroid carcinoma was significantly lower than adjacent tissues(0.384±0.028 vs 0.981±0.039,t=10.291,P<0.05);the expression of miR-126-3p in TPC-1 was the lowest among three types of thyroid cancer cells.Compared with NC group,the proliferation of TPC-1 in mimic group was significantly inhibited,the same with migration(26.68±4.48 vs 82.21±3.65,t=17.789,P<0.05)and invasion(12.28±1.03 vs 34.43±2.10,t=8.103,P<0.05),which the apoptosis was significantly increased[(15.32±3.20)% vs (8.12±1.17)%,t=4.623,P<0.05].Conclusion:The miR-126-3p expression is reduced in thyroid cancer tissue,overexpression of miR-126-3p significantly suppresses the proliferation,migration and invasion of thyroid cancer cells,and promotes the apoptosis,miR-126-3p can play an important biological function as a cancer suppressor gene in thyroid cancer.