Proliferative mechanism of microRNA-543 on non-small cell lung cancer cells by molecular biology
10.13699/j.cnki.1001-6821.2017.17.018
- VernacularTitle:用分子生物学方法检测微小RNA-543促进非小细胞肺癌细胞的增殖机制
- Author:
Yang CAI
1
;
Yong BAO
Author Information
1. 西南医科大学临床医学院
- Keywords:
microR-543;
proliferation;
apoptosis;
cell cycle;
progestin and adiponectin receptors 3
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(17):1671-1674
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the expression of microRNA-543 (miR-543) in non-small cell lung cancer (NSCLC) cell line A549,and further investigate the effects of niR-543 on cell proliferation,cell apoptosis,and cell cycle of A549 cells and underlying mechanism.Methods Quantitative real-time polymerase chain reaction was performed to detect the expression of microRNA-543 (miR-543) in NSCLC cell line A549 cells and normal lung epithehal ceils in BEAS-2B cells.The expression of miR-543 in A549 cells was silenced by miR-543 inhibitors.A549 cells were divided into miR-543 normal control (NC) group and miR-543 inhibitors group.Cell counting kit-8 was performed to detect the effects of miR-543 on cell proliferation.Flow cytometry was performed to detect the effects of miR-543 on cell apoptosis and cell cycle.Luciferase assays was performed to detect whether progestin and adiponectin receptors 3 (PAQR3) was the direct targets of miR-543 in A549 cells.Results The relative expression of miR-543 in normal human epithelial cells BEAS-2B was 1.00 ± 0.05,the relative expression of miR-543 in NSCLC cells A549 was 3.87 ± 0.04,the expression of miR-543 was significantly higher in the A549 cells than human normal lung epithehal ceils(P ＜ 0.05).A549 cells were transfected with miR-543 inhibitors,the relative expression of miR-543 in miR-543 normal control group was 1.00 ± 0.05,the relative expression of miR-543 in miR-543 inhibitors group was 0.43 ± 0.07,the expression of miR-543 inA549 cells was inhibited by miR-543 inhibitors (P ＜ 0.05).OD values at 1-5 d of miR-543 NC group were 0.25 ±0.05,0.48 ±0.04,0.73 ±0.06,0.98 ±0.05,1.35 ±0.04,OD values at 1-5 d of miR-543 inhibitors group were 0.25 ± 0.04,0.37 ± 0.04,0.48 ± 0.05,0.58 ± 0.05,0.66 ± 0.04,the cell proliferation was significantly inhibited by miR-543 inhibitors (P ＜ 0.05).The apoptosis rate of miR-543 NC group was (1.21 ±0.37)％,apoptosis of miR-543 inhibitors group was (5.78 ± 1.69)％,early apoptosis rate of miR-543 NC group was (0.84 ±0.21)％,early apoptosis rate of miR-543 inhibitors group was (2.18 ± 1.28) ％,cell apoptosis was significantly promoted by miR-543 inhibitors (P ＜ 0.05).G1 stage of miR-543 in NC group was (51.27 ± 3.19) ％,G1 stage of miR-543 inhibitors group was (59.30 ± 4.19) ％,cell cycle was arrested as G1 phase in A549 cells transfected with miR-543 inhibitors(P ＜0.05).The relative activity of luciferase reporter gene with wild type(WT) progestin and PAQR3 was 0.42 ±0.05,the relative activity of luciferase reporter gene with mutant PAQR3 was 0.97 ± 0.04,PAQR3 was the direct target of miR-543 (P ＜ 0.05) and miR-543 could direcdy regulate the expression of in A549 cells.Conclusion miR-543 could regulate the cell proliferation,cell apoptosis,and cell cycle of A549 by directly inhibiting the expression of PAQR3.
