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

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 PKM2 was analyzed by online database tabula muris. The proportion of apoptotic cells was detected by flow cytometry. The expression of PKM2 gene was detected by fluorescence quantitative PCR and Western blotting. PKM2 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 (P < 0.001). In addition, with the increase of amifostine concentration, PKM2 gene expression in HK-2 cells was increased in turn. The expression of PKM2 gene was increased to 8.76 times higher at 1 μmol/