Effect of targeted silencing of DNMT3A on collagen deposition, proliferation and migration activity of mouse lung fibroblasts
10.19405/j.cnki.issn1000-1492.2025.01.009
- Author:
Xianchen Wang
1
;
Junbo You
1
;
Hui Ling
1
;
Jiahao Fan
1
;
Qi Chen
2
;
Hui Tao
2
;
Jiming Sha
1
Author Information
1. Dept of Thoracic Surgery，The Second Affiliated Hospital of Anhui Medical University，Hefei 230601
2. Dept of Anesthesiology and Perioperative Medicine， The Second Affiliated Hospital of Anhui Medical University，Hefei 230601
- Publication Type:Journal Article
- Keywords:
DNMT3A;
DNA methylation;
lung fibroblasts;
proliferation;
migration;
pulmonary fibrosis
- From:
Acta Universitatis Medicinalis Anhui
2025;60(1):66-72
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs).
Methods:In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test.
Results:Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group.
Conclusion:Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.
- Full text:2025030416410566147靶向沉默DNMT3A对小鼠...沉积、增殖及迁移活性的影响_汪先晨.pdf
