3.5 Effects of uc.375 overexpression and FoxA1 overexpression on the function of MLE 12 cells
To further evaluate the effect of uc.375 overexpression and FoxA1 overexpression on MLE 12 cells function, MLE 12 cells were stably co-transfected with uc.375 overexpression /FoxA1 overexpression to overexpress uc.375 and FoxA1. CCK-8 assay was conducted and cell viability was decreased by FoxA1 overexpression (Figure 5A). In addition, the cell apoptosis was assessed by Hoechst/PI staining and flow cytometry. The cell apoptosis was notably increased by FoxA1 overexpression (Figure 5B and 5D). 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 reduced, whereas caspase3 and FoxA1 were markedly increased by FoxA1 overexpression (Figure 5C). In addition, the qRT-PCR results confirmed that uc.375 level was significantly elevated by uc.375 overexpression, the SP-C and UCP2 mRNA expression were inhibited, whereas FoxA1 was markedly promoted by FoxA1 overexpression (Figure 5E-H). Taken together, FoxA1 overexpression suppressed cell proliferation and stimulated apoptosis.