Lactate Dehydrogenase A Inhibits Mitophagy in Human Glioma Cells via AMPK Signaling Pathway
LIU Fei, HUANG Xueyang, YANG Mengting, ZHANG Junyao, FANG Wenjing, GONG Aihua*
The aim of the study was to investigate the effect of LDHA on mitophagy in human glioma cells. The sh-LDHA or sh-EGFP plasmids were transfected into the glioma cells U87MG, respectively. qRT-PCR and Western blot were used to detect the efficiency of plasmid knockdown. The expression levels of mitophagy-related proteins and AMPK signaling pathway-related proteins were detected by Western blot. Then mitochondrial ROS and mitochondrial membrane potential levels were detected by fluorescence staining. Compared with sh-EGFP group, the mRNA and protein levels of LDHA were significantly decreased, and mitochondrial ROS level raised, mitochondrial membrane potential declined in the sh-LDHA group. Also, the expression of mitophagy-related proteins PINK1, Parkin, BNIP3, BNIP3L and AMPK signaling pathway-related proteins p-AMPK were significantly increased, while p-mTOR was decreased under these processing conditions in the sh-LDHA group. In summary, LDHA inhibits AMPK signaling pathway, reducing mitochondrial ROS levels, enhancing mitochondrial membrane potential, and inhibits mitophagy.
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