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Fluid shear stress applied by orbital shaking induces MG-63 osteosarcoma cells to activate ERK in two phases through distinct signaling pathways.

ABSTRACT: Fluid shear stress (FSS) induces a series of biochemical responses in osteoblasts, and this "mechanoresponse" regulates their survival, proliferation and differentiation. However, the events in cells immediately after FSS application are unclear, and how biochemical signals from soluble factors modify the mechanoresponses is largely unknown. We used the orbital shaking method, instead of the frequently used parallel plate method, to examine activation of ERK and AKT by FSS for detailed tracking of its temporal transition. We found that ERK activation by orbital shaking was biphasic. The early phase was independent of Ca²⁺, PI3-kinase, and Rho kinase but required RAF activity. The late phase was dependent on Ca²⁺ but not RAF. These results suggest that the superior time-resolving capability of the orbital shaking method to separate the previously unrecognized Ca²⁺-independent early phase of ERK activation from the late phase. We also found that a certain combination of serum starvation and medium renewal affected ERK activation by FSS, suggesting that a soluble factor(s) may be secreted during serum starvation, which modified the phosphorylation level of ERK. These findings revealed novel aspects of the osteoblastic mechanoresponses and indicated that the orbital shaking method would be a useful, complementary alternative to the parallel plate method for certain types of study on cellular mechanoresponses.

SUBMITTER: Fukada T

PROVIDER: S-EPMC5614596 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Fluid shear stress applied by orbital shaking induces MG-63 osteosarcoma cells to activate ERK in two phases through distinct signaling pathways.

Fukada Takashi T Sakajiri Hiroki H Kuroda Mito M Kioka Noriyuki N Sugimoto Kenji K

Biochemistry and biophysics reports 20170111

Fluid shear stress (FSS) induces a series of biochemical responses in osteoblasts, and this "mechanoresponse" regulates their survival, proliferation and differentiation. However, the events in cells immediately after FSS application are unclear, and how biochemical signals from soluble factors modify the mechanoresponses is largely unknown. We used the orbital shaking method, instead of the frequently used parallel plate method, to examine activation of ERK and AKT by FSS for detailed tracking ...[more]

PMID: 28956013

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Dataset Information

Fluid shear stress applied by orbital shaking induces MG-63 osteosarcoma cells to activate ERK in two phases through distinct signaling pathways.

Publications

Fluid shear stress applied by orbital shaking induces MG-63 osteosarcoma cells to activate ERK in two phases through distinct signaling pathways.

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