Effect of CIP4 on human renal tubular epithelial to mesenchymal transition induced by transforming growth factor β1
10.3760/cma.j.issn.1001-7097.2011.04.013
- VernacularTitle:CIP4对转化生长因子β1诱导的人肾小管上皮细胞-间充质转分化的影响
- Author:
Shoujun BAI
;
Yamin ZHANG
;
Rui ZENG
;
Chuou XU
;
Lili LIU
;
Qiaodan ZHOU
;
Caixia LI
;
Guangchang PEI
;
Shuwang GE
;
Gang XU
;
Xiaocheng LIU
- Publication Type:Journal Article
- Keywords:
Transforming growth factor beta 1;
Kidney tubules;
Epithelial cells;
Epithelial to mesenchymal transition;
CIP4;
PI3K-Akt
- From:
Chinese Journal of Nephrology
2011;27(4):282-287
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the effect of CIP4(Cdc42 interacting protein 4)on human renal tubular epithelial to mesenchymal transition(EMT)induced by transforming growth factor β1(TGF-β1)and to study the associated mechanism. Methods Human proximal tubular epithelial cells (HK-2 cell line) were cultured with TGF-β1 (10μg/L) for 72 hours. The protein expressions of E-cadherin and α-SMA were measured by Western blotting. One set of siRNA oligos specific for CIP4 and CIP4 construction of the entire coding sequence were designed based on the full CIP4 sequence in GenBank. Then HK-2 cells were transfected with CIP4-siRNA or pcDNA3.1-hCIP4 via lipofactamine 2000. The protein expressions of CIP4, E-cadherin and α-SMA were evaluated respectively in control cells, TGF-β1 treated cells, siRNA transfected cells, pcDNA3.1-hCIP4-transfected cells by Western blotting. The distribution of E-cadherin and α-SMA was observed by confocal microscope. After TGF-β1-treated HK-2 cells were interferenced with specific inhibitor of PI3K-Akt (wortmannin) 1μmol/L for 48 hours, Western blotting was used to detect the CIP4 protein in control cells and interferenced cells. Results With TGF-β1 stimulation, the expression of E-cadherin protein was decreased markedly (P＜0.05), and in contract, the expression of α-SMA were increased notably (P＜0.05), which revealed that TGF-β1 could induce EMT. After transfected with CIP4-siRNA, the protein expression of E-cadherin was increased (P＜0.05), and the protein expression of α-SMA was decreased (P＜0.05). The EMT induced by TGF-β1 was effectively reversed. After transfected with pcDNA3.1-hCIP4, the expression of E-cadherin protein was down-regnlated (P＜0.05), and the expression of α-SMA protein was up-regulated compared with control group (P＜0.05), leading to EMT. After HK-2 cells were interferenced with wortmannin for 48 hours, the expression of CIP4 was decreased (P＜0.05). Conclusion TGF-β1 upregulates the expression of CIP4 via PI3K-Akt pathway, and CIP4 may participate in EMT induced by TGF-β1.
