Abstract
Lung cancer is a leading cause of death. Most previous studies have been based on traditional cell-culturing methods. However, lung cells are periodically subjected to mechanical forces during breathing. Understanding the mechanisms underlying the cyclic stretching induced in lung cells may be important for lung cancer therapy. Here, we applied cyclic stretching to stimulate the continual contraction that is present under physiological conditions in lung cells. We first uncovered the stretching-induced phosphoproteome in lung cancer cell line A549 and fibroblast cell line IMR-90. We identified 2048 and 2604 phosphosites corresponding to 837 and 1008 phosphoproteins in A549 and IMR-90, respectively. Furthermore, we combined our phosphoproteomics and public gene expression data to identify the biological functions in response to cyclic stretching. Interestingly, cytoskeletal and mitochondrial reorganization were enriched. We further used cell imaging analysis to validate the profiling results and found that this physical force changed cell alignment and mitochondrial length. This study not only reveals the molecular mechanism of cyclic stretching but also provides evidence that cell stretching causes cellular rearrangement and mitochondrial length change.
Highlights
Cyclic stretching is a process that can be investigated in adherent cells seeded on a flexible surface and stretched periodically
We found that gene expression, the negative regulation of biological processes, and intracellular signal transduction functions are unique in IMR-90 cells
We have provided a global view of the stretching-induced phosphoproteome by stretching lung cancer cell line A549 and fibroblast cell line IMR-90 for short periods (15, 30, or 60 min) and a longer period (24 h)
Summary
Cyclic stretching is a process that can be investigated in adherent cells seeded on a flexible surface and stretched periodically This procedure creates a dynamic environment that simulates that of continually contracting organs, such as the lungs, heart, and muscle tissue. Study, we applied different time scales of 2c.yRcelsiucltsstretching to gain insight into the stretching-induced phosphoproteome in lung cancer cell linTheeAex5p4e9rimaenndtalluwnorgkflfiobwroisbilllaussttralitnedeinIMFiRgu-r9e01. 10% CSA and a uniaxial stretching direction were commonly applied in cyclic stretched lung cells [22,23,24,25,26,27,28]. 10% CSA change and a uniaxial stretching direction are used for the cyclic stretching experiment
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