Negative pressure induces dedifferentiation of hepatocytes via RhoA/ROCK pathway

Biomechanical forces are known to regulate the biological behaviors of cells. Although negative pressure has been used for wound healing, it is still unknown about its role in regulating cell plasticity. We investigated whether negative pressure could induce the dedifferentiation of hepatocytes. Using a commercial device, we found that the exposure of primary human hepatocytes to -50 mmHg quickly induced the formation of stress fibers and obviously changed cell morphology in 72 h. Moreover, the exposure of hepatocytes to -50 mmHg significantly upregulated RhoA, ROCK1, and ROCK2 in 1-6 h, and dramatically enhanced the expression of marker molecules on stemness, such as OCT4, SOX2, KLF4, MYC, NANOG, and CD133 in 6-72 h. However, all these changes in hepatocytes induced by -50 mmHg stimulation were almost abrogated by ROCK inhibitor Y27623. Our data suggest that an appropriate force of negative pressure stimulation can effectively induce the dedifferentiation of hepatocytes via RhoA/ ROCK pathway activation. (c) 2023 Elsevier Inc. All rights reserved.

Automated summary

Biomechanical forces have the ability to regulate cell behaviors. This study investigated the role of negative pressure in regulating the dedifferentiation of hepatocytes. It was found that exposure to -50 mmHg pressure quickly induced stress fiber formation and changes in cell morphology. Furthermore, the activation of RhoA/ROCK pathway and the upregulation of stemness markers were observed. Inhibition of ROCK abrogated these changes induced by -50 mmHg pressure.