Effects of microRNA-16 on pulmonary surfactant associated protein of human alveolar epithelial cell A549 / 中国新生儿科杂志

ABSTRACT

Objective To study the regulatory role of microRNA-16 (miR-16) on human pulmonary surfactant associated protein (SP).Method Human alveolar epithelial A549 cells were transfected by miR-16 analogue,analogue negative control,inhibitor and inhibitor negative control.Blank control group was also set up at the same time.The proliferative abilities of the cells in each group were measured using cell counting kit-8 (CCK8) test.The expressions of miR-16,SP-A,SP-B and SP-C mRNA were examined using reverse transcription polymerase chain reaction (RT-PCR).The protein levels of SP-A,SP-B and SP-C were examined using western blotting method.Result RT-PCR showed that the expression of miR-16 after transfected with miR-16 analogue (34.11± 1.79) was higher than the negative control group (1.65 ± 1.07) and the blank control group (1.07 ±0.50).The expression of miR-16 after transfected with miR-16 inhibitor (0.36±0.05) was lower than the negative control group (0.96±0.13) and the blank control group (1.05±0.20).The differences were significant (all P＜0.05),and indicated that miR-16 over-expression and suppression were successfully achieved.Compared with the blank control group,cell proliferation at different time points in the analogue negative control group and the inhibitor negative control group showed no significant differences (all P＞0.05).Compared with the blank control group (1.02±0.19,1.01±0.09,1.01± 0.12) and the analogue negative control group (1.08±0.24,1.00±0.14,1.00±0.05),miR-16 down-regulated the mRNA expressions of SP-A,SP-B and SP-C (0.58±0.16,0.67±0.05,0.61±0.12).On the other hand,compared with the blank control group (1.02±0.19,1.01±0.09,1.01±0.12) and the inhibitor negative control group (1.05±0.22,0.99±0.13,0.98±0.10),miR-16 up-regulated the mRNA expressions of SP-A,SP-B and SP-C (1.66±0.33,1.29±0.11,1.23±0.12)(all P＜0.05).The trends of protein level of SP-A,SP-B and SP-C were related to their mRNA expression.Conclusion This study indicates that miR-16 inhibits pulmonary surfactant associated protein in A549 cells.