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1.
Sci Rep ; 6: 32418, 2016 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-27581365

RESUMO

This study focused on the effects of simulated microgravity (s-µg) on mechanical properties, major cytoskeleton biopolymers, and morphology of endothelial cells (ECs). The structural and functional integrity of ECs are vital to regulate vascular homeostasis and prevent atherosclerosis. Furthermore, these highly gravity sensitive cells play a key role in pathogenesis of many diseases. In this research, impacts of s-µg on mechanical behavior of human umbilical vein endothelial cells were investigated by utilizing a three-dimensional random positioning machine (3D-RPM). Results revealed a considerable drop in cell stiffness and viscosity after 24 hrs of being subjected to weightlessness. Cortical rigidity experienced relatively immediate and significant decline comparing to the stiffness of whole cell body. The cells became rounded in morphology while western blot analysis showed reduction of the main cytoskeletal components. Moreover, fluorescence staining confirmed disorganization of both actin filaments and microtubules (MTs). The results were compared statistically among test and control groups and it was concluded that s-µg led to a significant alteration in mechanical behavior of ECs due to remodeling of cell cytoskeleton.


Assuntos
Citoesqueleto de Actina/ultraestrutura , Células Endoteliais da Veia Umbilical Humana/ultraestrutura , Microtúbulos/ultraestrutura , Simulação de Ausência de Peso/instrumentação , Citoesqueleto de Actina/metabolismo , Fenômenos Biomecânicos , Forma Celular , Elasticidade , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Microtúbulos/metabolismo , Viscosidade , Simulação de Ausência de Peso/métodos
2.
J Biomech ; 48(6): 1172-8, 2015 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-25678199

RESUMO

Biomechanical behaviours of cells change during cancer progression due to alterations in the main cytoskeletal proteins. Microtubules play a vital role in mitosis and in supporting the integrity of the cell due to their ability to withstand high compressive loads. Accordingly, microtubule-targeting agents (MTAs) have become one of the most promising classes of drugs in cancer therapy. This study evaluated changes in visco-elastic parameters induced by an appropriate concentration of an antimitotic drug in two different grades of colon cancer cells. Actin microfilaments and microtubules contents in the cells were evaluated by Western blot analysis and fluorescence intensity calculation. Micropipette aspiration experiments showed that the MTA had distinct mechanical effects on different cell lines. The more aggressive the cells, the greater the reduction in elasticity and viscosity. Invasive cells had a higher initial instantaneous Young's modulus than primary cells, but this reduced to approximately one half of the values for primary cells after 48 h of drug treatment. A considerable association was seen between the changes in mechanical properties and the microtubule to F-actin microfilament content ratio, which decreased with MTA treatment.


Assuntos
Albendazol/farmacologia , Antimitóticos/farmacologia , Neoplasias do Colo/patologia , Citoesqueleto/efeitos dos fármacos , Actinas/metabolismo , Fenômenos Biomecânicos , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/metabolismo , Proteínas do Citoesqueleto/metabolismo , Citoesqueleto/metabolismo , Módulo de Elasticidade , Células HT29 , Humanos , Microtúbulos/metabolismo , Viscosidade
3.
J Biomech ; 47(2): 373-9, 2014 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-24315289

RESUMO

Biomechanical properties of cancer cells have been proposed as promising biomarkers to investigate cancer progression. Cytoskeletal reorganization alters these characteristics in different grades of cancer cells. In the present study based on the micropipette aspiration method, whole body evaluation for two different colon cancer cells was performed to determine viscoelastic parameters of the cells. A finite element model was developed for verification of experiments and predicting some behaviors of cells. Western blot analysis and fluorescence intensity for actin microfilaments and microtubules were performed to measure cell content of the proteins. It was illustrated that the proportion of microtubules and actin microfilaments is different in grade I and grade IV colon cancer cells in a manner that microtubules attain an effectual role in progressive reorganization of cytoskeleton in transition from nonaggressive to malignant phenotypes in cancer cells. Furthermore, it was concluded that larger instantaneous Young's modulus value for high grade cells is related to the existence of extensively build-up actin networks at the cell cortex. Based on the cell mechanics results, a simple parameter is suggested for sorting different grades of colon cancer cells.


Assuntos
Citoesqueleto de Actina/metabolismo , Neoplasias do Colo/patologia , Citoesqueleto/patologia , Microtúbulos/metabolismo , Actinas/metabolismo , Albendazol/farmacologia , Western Blotting , Movimento Celular/efeitos dos fármacos , Módulo de Elasticidade/fisiologia , Elasticidade , Análise de Elementos Finitos , Células HT29 , Humanos , Microtúbulos/patologia , Viscosidade
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