Expression of microRNA-646 in A549 cells treated by lipopolysaccharide and its function
Bin Wang, Dongliang Cheng, Linli Wang, Jiang Du
Abstract
Bin Wang, Dongliang Cheng, Linli Wang, Jiang Du
Abstract
Objective To measure the expression of microRNA-646( miR-646 ) in A549 cells under different concentrations of lipopolysaccharide(LPS) treatment, and to explore the possible mechanism of miR-646 in A549 cells under LPS treatment. Methods A549 cells were divided into control group and experimental groups.A549 cells from the control group were treated with RPMI-1640 and A549 cells from the experimental groups were treated with LPS(5 mg/L, 10 mg/L, 15 mg/L) in a duration of 24 hours.Immunocytochemical method and Western blot were used to detect the changes in surfactant protein A (SP-A) and surfactant protein C (SP-C), and quantitative real-time polyme-rase chain reaction was used to detect the changes in miR-646 in all groups. Results Compared with control group, the expression of SP-A in cytoplasm of A549 cells were decreased in experimental groups (all P<0.05), and LPS in diffe-rent concentrations induced the expression of SP-A in A549 cells in a dose-dependent manner.Compared with control group, the expression of SP-C in cytoplasm of A549 cells was decreased in experimental groups (all P<0.05). The expression of SP-C was the lowest in 10 mg/L LPS treatment group.The relative expression level of the control group of miR-646 was 0.9597±0.0200, in 5 mg/L LPS treatment group it was 1.6319±0.1325, in 10 mg/L LPS treatment group it was 2.4762±0.1380, and in 15 mg/L LPS treatment group it was 1.6642±0.0938.There were statistically significant differences between the experimental groups and control group (all P<0.01). Conclusion miR-646 may have a biological function in acute respiratory distress syndrome through inhibiting the transcription of SP-C. Key words: microRNA-646; Acute respiratory distress syndrome; Surfactant protein C; A549 cell
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Objective To measure the expression of microRNA-646( miR-646 ) in A549 cells under different concentrations of lipopolysaccharide(LPS) treatment, and to explore the possible mechanism of miR-646 in A549 cells under LPS treatment. Methods A549 cells were divided into control group and experimental groups.A549 cells from the control group were treated with RPMI-1640 and A549 cells from the experimental groups were treated with LPS(5 mg/L, 10 mg/L, 15 mg/L) in a duration of 24 hours.Immunocytochemical method and Western blot were used to detect the changes in surfactant protein A (SP-A) and surfactant protein C (SP-C), and quantitative real-time polyme-rase chain reaction was used to detect the changes in miR-646 in all groups. Results Compared with control group, the expression of SP-A in cytoplasm of A549 cells were decreased in experimental groups (all P<0.05), and LPS in diffe-rent concentrations induced the expression of SP-A in A549 cells in a dose-dependent manner.Compared with control group, the expression of SP-C in cytoplasm of A549 cells was decreased in experimental groups (all P<0.05). The expression of SP-C was the lowest in 10 mg/L LPS treatment group.The relative expression level of the control group of miR-646 was 0.9597±0.0200, in 5 mg/L LPS treatment group it was 1.6319±0.1325, in 10 mg/L LPS treatment group it was 2.4762±0.1380, and in 15 mg/L LPS treatment group it was 1.6642±0.0938.There were statistically significant differences between the experimental groups and control group (all P<0.01). Conclusion miR-646 may have a biological function in acute respiratory distress syndrome through inhibiting the transcription of SP-C. Key words: microRNA-646; Acute respiratory distress syndrome; Surfactant protein C; A549 cell
Key concepts: A549 cell, Lipopolysaccharide, Western blot, Molecular biology, Cytoplasm, Chemistry, Blot, Real-time polymerase chain reaction