pLP/VSVG

Here, researchers explored the regulatory mechanism of miR-10b on epithelial-mesenchymal transition (EMT) and its impact on the proliferation and migration of nasopharyngeal carcinoma cells. RT-qPCR detected the expression of miR-10b in nasopharyngeal carcinoma cell CNE1. The NP69 nasopharyngeal mucosal cell line was used to determine the expression of miR-10b after lentivirus infection. Studies have shown that miR-10b is highly expressed in CNE1 cells. The stable expression of miR-10b promotes the proliferation and migration of NP69 cells, downregulates the expression of epithelial cell markers E-cadherin and β-catenin, and upregulates the expression of mesenchymal cell markers fibronectin, N-cadherin, vimentin and MMP-9 resulting in cell EMT. Therefore, miR-10b can promote the proliferation and migration of nasopharyngeal carcinoma cells and induce EMT in nasopharyngeal carcinoma cells, which may become a new target for the treatment of nasopharyngeal carcinoma.

In this study, lentivirus-miR-10b infected NP69 cells. Construction of a lentiviral system expressing miR-10b. The sequence of miR-10b was inserted into the lentiviral plasmid pLP/VSVG. pLP/VSVG-miR-10b, pLP1 and pLP2 were co-transfected into 293 T cells, and the supernatant containing lentivirus was collected after 48 hours. NP69 cells were seeded in a 6-well plate, cultured overnight into a monolayer, and then incubated with lentivirus supernatant for 24 hours to allow lentivirus to infect the cells. Lentivirus-infected cells were collected on the day of infection and 2 to 6 days after infection, and RT-qPCR was used to detect the expression level of miR-10b. NP69 cells infected with blank pLP/VSVG were used as control.

Figure 1. Expression level of miR-10b after lentivirus infection of NP69 cells. RT-qPCR was used to detect the expression level of miR-10b on days 1, 2, 3, 4, 5, and 6 after lentivirus infection of NP69 cells. Blank pLP/VSVG is a virus-free control. (Wang W. 2017)