Jingjing Hu1, Lu Wang2, Caihong Guan3. 1. Department of Respiratory Medicine, Hangzhou Dingqiao Hospital, Hangzhou, China. 2. Department of Emergency, Hangzhou Dingqiao Hospital, Hangzhou, China. 3. Department of Respiratory, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou, China.
Abstract
Background: Studies showed that miR-532-5p suppresses proliferation and induces apoptosis of lung cancer (LC) cells; its role in LC is not fully understood. Therefore, this research aimed to reveal the effect and mechanism of miR-532-5p on migration and invasion of LC cells. Materials and Methods: The transfection efficiencies of miR-532-5p mimic, inhibitor, and overexpressed CCR4 were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The relationships between miR-532-5p and CCR4 in A549 and SBC-5 cells were predicted by targetScan and verified by dual-luciferase reporter assay, Western blot, and qRT-PCR. Migration and invasion of cells transfected with miR-532-5p mimic, inhibitor, and CCR4 were determined by scratch test and transwell assay, respectively. The levels of epithelial-to-mesenchymal transition (EMT)-related proteins (E-cadherin (E-Cad)), N-catenin (N-Cad), and vimentin) in cells were measured by Western blot. Results: MiR-532-5p mimic suppressed migration and invasion, while miR-532-5p inhibitor promoted migration and invasion of cells. CCR4 was a downstream target of miR-532-5p and both its protein and mRNA expressions were inhibited by miR-532-5p mimic, but promoted by miR-532-5p inhibitor. CCR4 promoted migration, invasion, and EMT process, and such effects of CCR4 were reversed by miR-532-5p mimic. Conclusion: MiR-532-5p functioned as a cancer suppressor by negatively regulating CCR4 in LC cells, pointing to a potential protective mechanism of miR-532-5p to LC patients.
Background: Studies showed that miR-532-5p suppresses proliferation and induces apoptosis of lung cancer (LC) cells; its role in LC is not fully understood. Therefore, this research aimed to reveal the effect and mechanism of miR-532-5p on migration and invasion of LC cells. Materials and Methods: The transfection efficiencies of miR-532-5p mimic, inhibitor, and overexpressed CCR4 were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The relationships between miR-532-5p and CCR4 in A549 and SBC-5 cells were predicted by targetScan and verified by dual-luciferase reporter assay, Western blot, and qRT-PCR. Migration and invasion of cells transfected with miR-532-5p mimic, inhibitor, and CCR4 were determined by scratch test and transwell assay, respectively. The levels of epithelial-to-mesenchymal transition (EMT)-related proteins (E-cadherin (E-Cad)), N-catenin (N-Cad), and vimentin) in cells were measured by Western blot. Results:MiR-532-5p mimic suppressed migration and invasion, while miR-532-5p inhibitor promoted migration and invasion of cells. CCR4 was a downstream target of miR-532-5p and both its protein and mRNA expressions were inhibited by miR-532-5p mimic, but promoted by miR-532-5p inhibitor. CCR4 promoted migration, invasion, and EMT process, and such effects of CCR4 were reversed by miR-532-5p mimic. Conclusion:MiR-532-5p functioned as a cancer suppressor by negatively regulating CCR4 in LC cells, pointing to a potential protective mechanism of miR-532-5p to LCpatients.