Molecular imaging of membrane proteins and microfilaments using atomic force microscopy

Jung, Se-Hui; Park, Donghyun; Park, Jae Hyo; Kim, Young Myeong; Ha, Kwon‐Soo

doi:10.3858/emm.2010.42.9.064

Cited by 20 publications

(15 citation statements)

References 72 publications

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“…Comparison with AFM, the resolution of fluorescence microscopy is lower, besides, the sample also need to fluorescent staining. However, AFM is reported that can be analyzed to get the parameters pertaining to the examined surface in a quantitative form at nanometer resolution (Jung et al , ). Although electron microscopy techniques such as transmission electron microscopy reaches nanometer and subnanometer resolution, the process of sample preparation needs complex and invasive treatments that could dramatically modify cell native structure (Tai and Tang, ; Daban, ).…”

Section: Introductionmentioning

confidence: 99%

Curcumin disturbed cell‐cycle distribution of HepG2 cells via cytoskeletal arrangement

et al. 2012

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Due to its extensive antitumor activity, curcumin has been focused on by more researchers. But, its antiproliferative mechanisms are still unknown. Here we studied the antiproliferative activity of curcumin in human liver cancer HepG2 cells. In order to analyze the cytotoxic activity and anticancer mechanisms of curcumin, we carried out cytotoxicity tests using 3-[4,5-dimethyl-2-thiazolyl]-2,5 diphenyltetrazolium bromide (MTT) assay. The HepG2 cell cycle distribution and the expression of tubulin were detected by flow cytometry. Alterations in morphological and cytoskeletal properties of HepG2 cells were investigated using atomic force microscopy (AFM). Simultaneously, the effects of curcumin on the growth and proliferation of HepG2 cells were also assayed by MTT method. Cells were incubated with different doses of curcumin (0-80 μmol/l) for 24 h, the cell viability decreased from 91.10 ± 3.2% to 10.84 ± 4.0%, and the 50% inhibiting concentration (IC50 ) was 23.15 ± 0.37 μmol/l. Moreover, flow cytometry quantitatively detected that curcumin treatment resulted in a dose-dependent accumulation of HepG2 cells in G2/M phase with concomitant losses from G0/G1 phase, so curcumin caused cell-cycle arrest at G2/M phase. Furthermore, we discovered that curcumin was able to upregulate the expression of tubulin in HepG2 cells. In addition, AFM analysis including cell-membrane structure and cytoskeleton networks is helpful to explain the relationship between the changes of cells and external pharmacologic stimulation.

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Section: Introductionmentioning

confidence: 99%

Curcumin disturbed cell‐cycle distribution of HepG2 cells via cytoskeletal arrangement

et al. 2012

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“…Atomic force microscope (AFM) is a powerful tool for the characterization of biological sample. It can not only obtain the high-resolution images of cell morphology, membrane ultrastructure, and membrane proteins (Jung et al, 2010) but also measure mechanical properties of cell membrane, such as elasticity (Lekka and Laidler, 2009) and adhesion (Hosseini et al, 2009). In this study, HUVECs were plated and cultured on PLL-coated plate and FN-coated plate with or without celastrol treatment, and then morphology, cell surface roughness and cell mechanics were detected by AFM.…”

Section: Introductionmentioning

confidence: 99%

AFM studied the effect of celastrol on β1 integrin-mediated HUVEC adhesion and migration

Chen

Jin

Cai

2012

SCA

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Integrin-mediated human umbilical vein endothelial cells (HUVECs) adhesion to the extracellular matrix plays a fundamental role in tumor-induced angiogenesis. Celastrol, a traditional Chinese medicine plant, has possessed anticancer and suppressed angiogenesis activities. Here, the mechanism underling the antiangiogenesis capacity of celastrol was investigated by exploring the effect of celastrol on β1(CD29) integrin-mediated cell adhesion and migration. Flow cytometry results showed that the HUVECs highly expressed CD29 and cell adhesion assay indicated that celastrol specifically inhibited the adhesion of HUVECs to fibronectin (FN) without affecting nonspecific adhesion to poly-L-lysine (PLL). After cell FN adhesion being inhibited, the cell surface nanoscale structure and adhesion force were detected by atomic force microscope (AFM). High-resolution imaging revealed that cell morphology and ultrastructure changed a lot after being treated with celastrol. The membrane average roughness (Ra) and the major forces were decreased from 31.34 ± 4.56 nm, 519.60 ± 82.86 pN of 0 μg/ml celastrol to 18.47 ± 6.53 nm, 417.79 ± 53.35 pN of 4.0 μg/ml celastrol, 10.54 ± 2.85 nm, 258.95 ± 38.98 pN of 8.0 μg/ml celastrol, respectively. Accompanying with the decrease of adhesion force, the actin cytoskeleton in the cells was obviously disturbed by the celastrol. All of these changes influenced the migration of HUVECs from the wound-healing migration assay. Taken together, our results suggest that celastrol can be as an inhibitor of HUVEC adhesion to FN. This work provides a novel approach to inhibition of tumor angiogenesis and tumor growth.

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“…AFM images provide detailed morphological features such as size, shape and surface topography at sub-nanometer resolutions [54][55][56][57]. In addition, it provides information on cellular architectures such as structural, conformational, and constitutional information of cytoskeletal proteins and membrane lipids [58][59][60][61]. During pathological progression, cells often undergo morphological modulations.…”

Section: Morphologymentioning

confidence: 99%

Nano-scientific Application of Atomic Force Microscopy in Pathology: from Molecules to Tissues

Kiio

Park

2020

Int. J. Med. Sci.

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The advantages of atomic force microscopy (AFM) in biological research are its high imaging resolution, sensitivity, and ability to operate in physiological conditions. Over the past decades, rigorous studies have been performed to determine the potential applications of AFM techniques in disease diagnosis and prognosis. Many pathological conditions are accompanied by alterations in the morphology, adhesion properties, mechanical compliances, and molecular composition of cells and tissues. The accurate determination of such alterations can be utilized as a diagnostic and prognostic marker. Alteration in cell morphology represents changes in cell structure and membrane proteins induced by pathologic progression of diseases. Mechanical compliances are also modulated by the active rearrangements of cytoskeleton or extracellular matrix triggered by disease pathogenesis. In addition, adhesion is a critical step in the progression of many diseases including infectious and neurodegenerative diseases. Recent advances in AFM techniques have demonstrated their ability to obtain molecular composition as well as topographic information. The quantitative characterization of molecular alteration in biological specimens in terms of disease progression provides a new avenue to understand the underlying mechanisms of disease onset and progression. In this review, we have highlighted the application of diverse AFM techniques in pathological investigations.

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Molecular imaging of membrane proteins and microfilaments using atomic force microscopy

Cited by 20 publications

References 72 publications

Curcumin disturbed cell‐cycle distribution of HepG2 cells via cytoskeletal arrangement

Curcumin disturbed cell‐cycle distribution of HepG2 cells via cytoskeletal arrangement

AFM studied the effect of celastrol on β1 integrin-mediated HUVEC adhesion and migration

Nano-scientific Application of Atomic Force Microscopy in Pathology: from Molecules to Tissues

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