Functional activity of mitochondria associated with cytoskeleton stability in the HEK293

Abstract

Mitochondria are dynamic cellular organelles that provide the biological energy to the cell. The dynamic property of the mitochondria is associated with the cytoskeleton network in the cell. However, relationship is no clear identified between cytoskeleton instability and the functional activity of mitochondria in cell. This study is designed to assess the dynamic properties of the mitochondria, and specifically to observe whether they are associated with the cytoskeleton stability for the mitochondria’s functional activityWe prepared the cytoskeleton stability regulation model with the HEK293 cell line by vinorelbine which microtubule distributor and the cytoskeleton stabilizer as a taxol. We determined the optimal concentration of vinorelbine and taxol though cytotoxicity assay with the HEK293 cell line. Then, we analyzed the functional activity of the mitochondria depending on the microtubule instability. First, we undertook an observation of cell morphology under distributor or stabilizer. We analyzed the metabolic activity of the cell with the CCK and ATP productive assay. And we performed confocal live imaging for observation of mitochondria morphology in the cytoskeleton instable cells. We performed the reverse transcription polymer chain reaction for analysis of the mRNA expression in the cytoskeleton’s instable cells with Tau protein, Glut1 and GAPDH as the normalization marker.The vinorelbine treated cell shows dramatically changes with cell morphology compared with control. And the vinorelbine treated cell significantly increases the fission ratios of the mitochondria compared with the original form based on the confocal images. Microtubule instable cells significantly decreased the ATP’s productive ratios and the functional activity of the mitochondria. However, the cytoskeleton stabilizer as a taxol(0.01uM) combined with vinorelbine treated cells, shows recovery effect to cell morphology recovery effect like the control cells that were only treated with vinorebine. And taxol that was combined with vinorelbine treated cells significantly increased bioenergy product ratios and its functional activity compared with the vinorelbine.In conclusion, the dynamic mitochondrial motility might influence the microtubule stability in cells. Because, vinorelbine‐treated cells displayed destabilized microtubules, loss of motility and fission formation of mitochondria. And loss motility of the mitochondria shows poor ATP productive ratios, resulting in a phenotype like that observed in the cells. This data suggests that functional activity of the mitochondria is associated with the microtubule stability in the cells. And microtubule stabilizer has a possibility to recovery functional activity in the microtubule instable cell like aging process. This finding can be applying to understand the mechanism of aging.Support or Funding InformationThis research was supported by the University Innovation Support Project (UISP) through the National Research Foundation (NRF) funded by MEST(Project No. 20190104)CCK analysis(A) and cell morphology(B) of NaB and Taxol treated cellsFigure 1ATP productive ratios(A) and fission form of mitochondria(B) of cellsFigure 2