3.4 Effects of uc.375 silencing and FoxA1 silencing on the function of MLE 12 cells
To further evaluate the effect of uc.375 silencing and FoxA1silencing on MLE 12 cells function, MLE 12 cells were stably co-transfected with uc.375 silencing/FoxA1 silencing to knock down uc.375 and FoxA1. CCK-8 assay was conducted and cell viability was increased by FoxA1 silencing (Figure 4A). In addition, the cell apoptosis was assessed by Hoechst/PI staining and flow cytometry. The cell apoptosis was notably reduced by FoxA1 silencing (Figure 4B and 4D). Moreover, the protein levels of caspase3, FoxA1, SP-C and UCP2 were detected by western blot in MLE12 cells. The results showed that SP-C and UCP2 were increased, whereas caspase3 and FoxA1 were markedly inhibited by FoxA1 silencing (Figure 4C). In addition, the qRT-PCR results confirmed that uc.375 level was significantly reduced by uc.375 silencing, the SP-C and UCP2 mRNA expression were stimulated, whereas FoxA1 was markedly suppressed by FoxA1 silencing (Figure 4E-H). Taken together, FoxA1 silencing stimulated cell proliferation and suppressed apoptosis.