Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2

Rong XUE; Jingang MA; Yingping LI; Jian ZHANG; Peijuan GAO; Wenhui HUANG; Xiaojun YANG; Xiaoli LI

doi:10.16016/j.2097-0927.202207159

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Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2

Basic Medicine | 更新时间：2026-01-09

- Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2
- Journal of Army Medical University Vol. 45, Issue 5, Pages: 433-440(2023)
- 作者机构：
  
  1.730000 兰州，甘肃省人民医院肾内科
  2.730000 兰州，甘肃省人民医院普外科
  3.730050 兰州，西宁联勤保障中心药品仪器监督检验站
- 作者简介：
- 基金信息：
  
  the Regional Science Fund Program of National Natural Science Foundation of China(81660398);the Natural Science Foundation of Gansu Province(21JR7RA625);the Natural Science Foundation of Gansu Province(22JR11RA256)
- DOI：10.16016/j.2097-0927.202207159
  CLC： R322.61;R966;R979.19
- Received：25 July 2022，
  
  Revised：2022-09-16，
  
  Published：15 March 2023
- 稿件说明：
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Rong XUE, Jingang MA, Yingping LI, et al. Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2[J]. Journal of Army Medical University, 2023, 45(5): 433-440.
DOI：

Rong XUE, Jingang MA, Yingping LI, et al. Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2[J]. Journal of Army Medical University, 2023, 45(5): 433-440. DOI： 10.16016/j.2097-0927.202207159.

摘要

目的

探究氨磷汀缓解顺铂肾毒性的分子机制。

方法

在肾近端小管上皮细胞系HK-2及原代细胞中，采用CCK-8细胞活性实验检测氨磷汀对顺铂细胞毒性的抑制作用。通过在线数据库tabula-muris分析肾脏组织各细胞亚群

PKM

2的表达。通过流式细胞术检测细胞凋亡比例。通过荧光定量PCR和蛋白免疫印迹实验检测细胞中

PKM

2基因表达。通过小RNA干扰技术敲低细胞中

PKM

2基因表达。

结果

氨磷汀显著增强HK-2细胞和原代肾近端小管上皮细胞抵抗顺铂的毒性。在顺铂处理的细胞中，随着氨磷汀浓度的增加，细胞的活力从0.37增加到0.45至0.77(

0.001)。另外，随着氨磷汀浓度的增加，HK-2细胞中

PKM

2基因表达依次增加，在1 μmol/L浓度时增加到8.76倍(

0.001)。在HK-2细胞中干扰

PKM

2基因表达，氨磷汀对顺铂的细胞毒性的保护作用显著受到抑制。顺铂诱导的细胞凋亡比例、标志蛋白Cleaved-PARP和Cleaved-caspase3的表达被氨磷汀显著抑制。单细胞数据分析的结果表明，相对于肾脏组织其他细胞亚群，

PKM

2在肾近端小管上皮细胞中表达最低。

结论

氨磷汀通过上调肾近端小管上皮细胞

PKM

2的表达抑制顺铂诱导的细胞凋亡，从而缓解顺铂的细胞毒性。

Abstract

Objective

To explore the molecular mechanism of amifostine in relieving cisplatin nephrotoxicity.

Methods

CCK-8 assay was used to detect the inhibitory effect of amifostine on cisplatin cytotoxicity in HK-2 and primary renal proximal tubular epithelial cells. The expression of

PKM

2 was analyzed by online database tabula muris. The proportion of apoptotic cells was detected by flow cytometry. The expression of

PKM

2 gene was detected by fluorescence quantitative PCR and Western blotting.

PKM

gene expression was knocked down by small RNA interference.

Results

Amifostine significantly enhanced the resistance of renal proximal tubular epithelial cells to cisplatin toxicity. In cisplatin treated cells

with the increase of the concentration of amifostine

the cell viability was increased from 0.37 to 0.45 to 0.77 (

0.001). In addition

with the increase of amifostine concentration

PKM

2 gene expression in HK-2 cells was increased in turn. The expression of

PKM

2 gene was increased to 8.76 times higher at 1 μmol/L of amifostine (

0.001).

PKM

2 gene expression in HK-2 cells was increased with the increase of amifostine concentration. When

PKM

2 gene expression was depleted in HK-2 cells

and the protective effect of amifostine on cisplatin cytotoxicity was significantly inhibited. The proportion of apoptotic cells was reduced and the levels of apoptotic markers Cleaved-PARP and Cleaved-Caspase3 induced by cisplatin were significantly inhibited by amifostine. Single-cell data analysis indicated that the expression of

PKM

2 in renal proximal tubular epithelial cells was the lowest compared with other cell subsets in renal tissue.

Conclusion

Amifostine inhibits cell apoptosis induced by cisplatin by up-regulating

PKM

2 in renal proximal tubular epithelial cells

and thus alleviates the cytotoxicity of cisplatin.

关键词

Keywords

references

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