Changes in cytoskeleton proteins and mechanical properties of the cells on soft substrates

doi:10.3969/j.issn.2095-4344.0882

Abstract

Abstract:

BACKGROUND: Preliminary studies have shown that substrate hardness is directly involved in cell regulation, but this effect can be hindered via suppression of myosin expression. Therefore, the effect of substrate hardness on myosin expression needs to be further quantified.

OBJECTIVE: To quantify the effect of substrate hardness on the distribution and movement rate of myosin II as well as on the distribution of actin fibers.

METHODS: Polyacrylamide gels were used to prepare substrates with hardness of 1, 10, 150 kPa. Cervical cancer were Hela cells stably transfected with fluorescent myosin II, and these transfected cells were incubated onto the substrates of different hardness. The fluorescence distribution of fluorescent myosin II along the long axis of Hela cells on the substrates of different hardness was measured. The movement rate of fluorescent myosin II was determined by fluorescence photobleaching recovery technique. F-actin distribution was observed using fluorescent staining. The elastic modulus of single cervical cancer cells spreading on different substrates was determined using micropipette aspiration technique.

RESULTS AND CONCLUSION: For the cells growing on the 150 kPa substrate, the highest intensity of fluorescent myosin II was located on the cell edge. For the cells on the substrates of 1 and 10 kPa, there was a reduced distribution of fluorescent myosin II on the cell edge. For the cells on the 150 kPa substrate, the recovery rate of fluorescent myosin II was significantly faster than that on the substrates of 1 and 10 kPa. Actin fibers in the cells growing on the substrates of 1, 10 and 150 kPa shared similarity in the overall fluorescence intensity. The elastic modulus of Hela cells on the substrate of 150 kPa was significantly higher than that on 1 kPa and 10 kPa substrates. Experimental results show that substrate hardness can significantly alter the regional effects of cytoskeletal proteins such as myosin and actin, and affect the cell status of cancer cells in different microenvironments.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Myosins, Actins, Elastic Modulus, Tissue Engineering

CLC Number:

R318

Hou Tian, An Mei-wen, Wang Li . Changes in cytoskeleton proteins and mechanical properties of the cells on soft substrates[J]. Chinese Journal of Tissue Engineering Research, doi: 10.3969/j.issn.2095-4344.0882.

Figures/Tables 5

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