TSSC3 suppresses fibrosis of renal tubular epithelium by regulating PI3K/Akt signaling pathway and anoikis resistance in myofibroblasts

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TSSC3 suppresses fibrosis of renal tubular epithelium by regulating PI3K/Akt signaling pathway and anoikis resistance in myofibroblasts

Basic Medicine | 更新时间：2026-01-09

- TSSC3 suppresses fibrosis of renal tubular epithelium by regulating PI3K/Akt signaling pathway and anoikis resistance in myofibroblasts
- Journal of Army Medical University Vol. 44, Issue 5, Pages: 421-431(2022)
- 作者机构：
  
  400042 重庆，陆军特色医学中心肾脏内科
- 作者简介：
- 基金信息：
  
  the General Program of Natural Science Foundation of Chongqing(cstc2020jcyj-msxmX0013)
- DOI：10.16016/j.2097-0927.202107152
  CLC： R163;R322.12;R329.25
- Received：24 July 2021，
  
  Revised：2021-10-17，
  
  Published：15 March 2022
- 稿件说明：
移动端阅览
Xinghong LIU, Yan CHEN, Fei XIAO, et al. TSSC3 suppresses fibrosis of renal tubular epithelium by regulating PI3K/Akt signaling pathway and anoikis resistance in myofibroblasts[J]. Journal of Army Medical University, 2022, 44(5): 421-431.
DOI：

Xinghong LIU, Yan CHEN, Fei XIAO, et al. TSSC3 suppresses fibrosis of renal tubular epithelium by regulating PI3K/Akt signaling pathway and anoikis resistance in myofibroblasts[J]. Journal of Army Medical University, 2022, 44(5): 421-431. DOI： 10.16016/j.2097-0927.202107152.

摘要

目的

2

研究印记基因STF cDNA 3(TSSC3)对肾脏来源的肌成纤维细胞(myofibroblast，MyoFb)抗失巢凋亡和促纤维化能力的影响及其相关机制。

方法

2

使用TGF-β1处理人肾近曲小管上皮细胞系HK-2诱导其成为MyoFb，并采用超低粘附板悬浮培养法以及外源性RGD多肽培养法建立失巢凋亡模型；使用含或不含TGF-β1的细胞培养基处理稳定转染过表达TSSC3慢病毒的HK-2细胞，采用流式细胞术及CCK-8检测TSSC3对MyoFb及HK-2细胞失巢凋亡、增殖的影响；qRT-PCR及Western blot检测PI3K/AKT介导的凋亡途径以及纤维化相关分子mRNA及蛋白表达情况。

结果

2

TGF-β1明显增加HK-2中α-SMA的表达、降低Villin的表达(

P

<

0.05)，且具有时间依赖性，表明TGF-β1成功诱导HK-2细胞成为MyoFb，且MyoFb的失巢凋亡率明显低于HK-2细胞(

P

<

0.01)，表明TGF-β1诱导的MyoFb具有失巢凋亡抵抗的能力。相对于HK-2细胞，MyoFb增殖率明显增加、失巢凋亡率明显降低(

P

<

0.01)，PI3K/AKT通路抑制剂LY204002则部分逆转了上述改变。过表达TSSC3明显抑制失巢凋亡模型中MyoFb中p-PI3K、p-AKT的表达，增加cleaved-caspase 3的表达。过表达TSSC3明显抑制MyoFb和HK-2细胞中Ⅰ型胶原、Ⅲ型胶原、纤连蛋白、PAI-1、MMP-2、MMP-9的mRNA及蛋白的表达。

结论

2

TSSC3通过下调PI3K/Akt途径抑制肾脏来源MyoFb的失巢凋亡抵抗及促纤维化的能力。

Abstract

Objective

2

To investigate the effects of tumor suppressing STF cDNA 3 (TSSC3) on the anoikis resistance and profibrogenic ability of renal myofibroblasts (MyoFb) and its related mechanisms.

Methods

2

Renal tubular epithelial HK-2 cells were stimulated with transforming growth factor beta 1 (TGF-β1) to induce the formation of MyoFb. The model of cell anoikis was established by culturing cells in ultra-low adhesion plate suspension or by exogenous RGD-containing peptides culture. In addition

HK-2 cells transfected with TSSC3-overexpressing lentiviral vector were treated with or without TGF-β1

and the effects of TSSC3 on the anoikis and proliferation of cells were observed using flow cytometry and cell counting kit-8 (CCK-8) assay. Finally

qRT-PCR and Western blotting were performed to detect the mRNA and protein expression of the PI3K/AKT pathway and fibrosis related molecules.

Results

2

The results demonstrated that α-SMA expression was significantly increased while that of Villin was decreased in HK-2 cells after stimulation of TGF-β1 in a time-dependent manner (

P

<

0.05)

indicating that HK-2 cells were successfully induced to MyoFb

which showed remarkably lower anoikis rate and higher proliferation than HK-2 cells (

P

<

0.01)

and had anoikis resistance. However

the above changes were partially reversed by PI3K/Akt pathway inhibitor LY294002. Further study suggested that overexpression of TSSC3 obviously inhibited the mRNA and protein expression of fibrosis related genes

such as typeⅠand type Ⅲ collagen

fibronectin

PAI-1

MMP-2 and MMP-9 in MyoFb and HK-2 cells. TSSC3 overexpression also up-regulated the level of cleaved-caspase 3 and attenuated the expression of p-PI3K and p-Akt in MyoFb.

Conclusion

2

TSSC3 suppresses the anoikis resistance and profibrogenic ability of renal myofibroblasts by down-regulating the PI3K/Akt pathway.

关键词

Keywords

references

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