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[The influence of miR-21 on HK-2 EMT cells induced by TGF-beta 1]
Yue-Qin Cai1, Yan-Qing Chu1, Ke-Yan Zhu1
1Zhejiang Chinese Medical University, Laboratory Animal Research Center/Comparative Medicine Institude, Hangzhou 310053, China.
Objective:
To investigate the effect of the miR-21 and its target mRNA in renal tubular epithelial mesenchymal transformation (EMT) model induced by transformation growth factor-β1(TGF-β1) in human renal tubular epithelial (HK-2) cells.
Methods:
HK-2 cells were divided into 6 groups:normal control group, TGF-β1 group, miR-21 mimic negative group, miR-21 mimic group, miR-21 inhibitor negative group and miR-21 inhibitor group. EMT model was established in HK-2 cells induced by 4 ng/ml TGF-β1. The level of miR-21, the mRNA and protein expression of EMT related factors were detected. MiR-21 mimic plasmid and miR-21 inhibitor plasmid were transfected into HK-2 cells that treated with TGF-β1 respectively using liposome transfection technique. Observe the impact of overexpression or inhibition expression of miR-21 on the mRNA and protein expression of EMT related factors and PTEN.
Results:
①Compared with the normal group, the level of miR-21 was significantly increased in model group (P<0.05), the mRNA and protein expression levels of epithelial cells marker E-cadherin was significantly decreased (P<0.01), while the mRNA and protein levels of mesenchymal cells marker α-SMA was significantly increased (P<0.05,P<0.01). ②Compared with the miR-21 mimic negative group, the level of miR-21 in miR-21 mimic group increased significantly (P<0.01), the mRNA and protein expression levels of PTEN and E-cadherin decreased significantly (P<0.05,P<0.01), the mRNA and protein levels of α-SMA increased significantly (P<0.05,P<0.01). Compared with the miR-21 inhibitor negative control group, the level of miR-21 in miR-21 inhibitor group decreased significantly (P<0.01), the mRNA and protein expression levels of PTEN and E-cadherin increased significantly (P<0.05,P<0.01), the mRNA and protein levels of α-SMA decreased significantly (P<0.05,P<0.01).
Conclusions:
MiR-21 may play an important role in EMT induced by TGF-β1 in HK-2 cells and regulate the expression of EMT related factors its target gene PTEN.
Insights
MicroRNA-21 (miR-21) significantly impacts renal tubular epithelial mesenchymal transformation (EMT) induced by TGF-β1. Modulating miR-21 levels affects EMT markers and PTEN expression in HK-2 cells.
Area of Science:
- Molecular Biology
- Cell Biology
- Renal Pathophysiology
Background:
- Renal tubular epithelial mesenchymal transformation (EMT) is a key process in kidney fibrosis.
- MicroRNAs (miRNAs) are emerging as critical regulators in various cellular processes, including EMT.
- Transforming growth factor-β1 (TGF-β1) is a potent inducer of EMT.
Purpose of the Study:
- To investigate the role of miR-21 in TGF-β1-induced EMT in human renal tubular epithelial (HK-2) cells.
- To determine the effect of miR-21 on the expression of EMT-related factors and its target gene PTEN.
Main Methods:
- HK-2 cells were treated with TGF-β1 to induce an EMT model.
- Cells were transfected with miR-21 mimic or inhibitor plasmids.
- Expression levels of miR-21, EMT markers (E-cadherin, α-SMA), and PTEN were analyzed at mRNA and protein levels.
Main Results:
- TGF-β1 treatment significantly increased miR-21 levels and induced EMT markers (decreased E-cadherin, increased α-SMA).
- Overexpression of miR-21 further promoted EMT and decreased PTEN and E-cadherin expression.
- Inhibition of miR-21 reversed EMT, increased E-cadherin and PTEN expression, and decreased α-SMA.
Conclusions:
- MiR-21 plays a significant role in TGF-β1-induced EMT in HK-2 cells.
- MiR-21 regulates the expression of EMT-related factors and its target gene PTEN during this process.
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