3.2 The effect of uc.375 silencing on MLE 12 cells function and FoxA1 expression
To assess the effect of uc.375 silencing on MLE 12 cells function, MLE 12 cells were stably transfected with uc.375 silent lentiviral vector to knock down uc.375. CCK-8 assay was conducted and cell viability was diminished by sh-uc.375 (Figure 2A). In addition, the effect of sh-uc.375 on cell apoptosis was assessed by Hoechst/PI staining and flow cytometry. The cell apoptosis was remarkably promoted by sh-uc.375 (Figure 2B and 2D). Moreover, the protein levels of caspase3, FoxA1, SP-C and UCP2 were investigated by western blot in MLE12 cells. The results showed that SP-C and UCP2 were suppressed, whereas caspase3 and FoxA1 were markedly stimulated by sh-uc.375 (Figure 2C). In addition, the qRT-PCR results confirmed that uc.375 level was significantly repressed by sh-uc.375, the SP-C and UCP2 mRNA expression were suppressed, whereas FoxA1 was markedly stimulated by sh-uc.375(Figure 2E-H). Taken together, uc.375 silencing inhibited cell proliferation and promoted apoptosis.