TSSC3通过调控PI3K/Akt-细胞失巢凋亡途径抑制肾小管上皮的纤维化表型

刘兴红; 陈雁; 肖菲; 戴欢子

doi:10.16016/j.2097-0927.202107152

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TSSC3通过调控PI3K/Akt-细胞失巢凋亡途径抑制肾小管上皮的纤维化表型

基础医学 | 更新时间：2026-01-09

- TSSC3通过调控PI3K/Akt-细胞失巢凋亡途径抑制肾小管上皮的纤维化表型
- TSSC3 suppresses fibrosis of renal tubular epithelium by regulating PI3K/Akt signaling pathway and anoikis resistance in myofibroblasts
- 陆军军医大学学报 2022年44卷第5期页码：421-431
- 作者机构：
  
  400042 重庆，陆军特色医学中心肾脏内科
- 作者简介：
  
  [ 戴欢子，E-mail：dhz084426@163.com DAI Huanzi, E-mail: dhz084426@163.com
- 基金信息：
  
  重庆市自然科学基金面上项目(cstc2020jcyj-msxmX0013)
- DOI：10.16016/j.2097-0927.202107152
  中图分类号： R163;R322.12;R329.25
- 收稿：2021-07-24，
  
  修回：2021-10-17，
  
  纸质出版：2022-03-15
- 稿件说明：
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刘兴红, 陈雁, 肖菲, 等. TSSC3通过调控PI3K/Akt-细胞失巢凋亡途径抑制肾小管上皮的纤维化表型[J]. 陆军军医大学学报, 2022,44(5):421-431.

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.
刘兴红, 陈雁, 肖菲, 等. TSSC3通过调控PI3K/Akt-细胞失巢凋亡途径抑制肾小管上皮的纤维化表型[J]. 陆军军医大学学报, 2022,44(5):421-431. DOI： 10.16016/j.2097-0927.202107152.

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

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

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

TGF-β1明显增加HK-2中α-SMA的表达、降低Villin的表达(

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(

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(

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

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

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

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

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 (

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 (

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

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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