• Molecules and Cells
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• v.41 no.12
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• pp.1045-1051
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• 2018

The developmentally regulated GTP binding protein 2 (DRG2) is involved in the control of cell growth and differentiation. Here, we demonstrate that DRG2 regulates microtubule dynamics in HeLa cells. Analysis of live imaging of the plus-ends of microtubules with EB1-EGFP showed that DRG2 deficiency (shDRG2) significantly reduced the growth rate of HeLa cells. Depletion of DRG2 increased 'slow and long-lived' subpopulations, but decreased 'fast and short-lived' subpopulations of microtubules. Microtubule polymerization inhibitor exhibited a reduced response in shDRG2 cells. Using immunoprecipitation, we show that DRG2 interacts with tau, which regulates microtubule polymerization. Collectively, these data demonstrate that DRG2 may aid in affecting microtubule dynamics in HeLa cells.

• The Korea Journal of Herbology
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• v.34 no.3
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• pp.55-61
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• 2019

Objectives : Recently, natural bioactive components catch a major attention for their potent anticarcinogenic activity. In this study, the inhibitory effect of Cinnamomi Cortex (CC) was examined in PC3 prostate cancer cells. Methods : The toxicity of CC extract was evaluated with cell viability and cell morphology. The activity of Yes associated protein (YAP) was tested with qRT-PCR for the target gene expression such as CTGF and AMOTL2. Western blotting was performed for the evaluation of phospho-YAP level. For cell motility analysis, cellular motility was imaged by live imaging system for 6 hr. Successive images were used for the generation of movie file. Using this movie file, cellular migration was manually tracked and analyzed using time-lapse microscope and Fiji software. Results : Cytotoxicity of CC extract was not detected at $500{\mu}g/m{\ell}$ or below concentration. Although $500{\mu}g/m{\ell}$ of CC extract reduced CTGF and AMOTL2 gene expression as YAP target genes, it was not statistically significant (CTGF expression P=0.0605, AMOTL2 expression P=0.4478). However, phosphorylated YAP was highly enhanced by CC extract treatment, when normalized with total YAP protein expression, suggesting YAP activation was inhibited. Finally prostate cancer cell motility was markedly reduced by $500{\mu}g/m{\ell}$ of CC extract. Conclusions : CC extract suppresses cancer cell motility and migration ability through inhibiting YAP activation without prostate cancer cell death, suggesting that this herb might be effective therapeutic drug for prostate cancer metastasis.

• Journal of Ginseng Research
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• v.48 no.2
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• pp.171-180
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• 2024

Background: Epimers of ginsenoside Rg3 (Rg3) have a low bioavailability and are prone to deglycosylation, which produces epimers of ginsenoside Rh2 (S-Rh2 and R-Rh2) and protopanaxadiol (S-PPD and R-PPD). The aim of this study was to compare the efficacy and potency of these molecules as anti-cancer agents. Methods: Crystal violet staining was used to study the anti-proliferatory action of the molecules on a human epithelial breast cancer cell line, MDA-MB-231, and human umbilical vein endothelial cells (HUVEC) and compare their potency. Cell death and cell cycle were studied using flow cytometry and mode of cell death was studied using live cell imaging. Anti-angiogenic effects of the drug were studied using loop formation assay. Molecular docking showed the interaction of these molecules with vascular endothelial growth factor receptor-2 (VEGFR2) and aquaporin (AQP) water channels. VEGF bioassay was used to study the interaction of Rh2 with VEGFR2, in vitro. Results: HUVEC was the more sensitive cell line to the anti-proliferative effects of S-Rh2, S-PPD and R-PPD. The molecules induced necroptosis/necrosis in MDA-MB-231 and apoptosis in HUVEC. S-Rh2 was the most potent inhibitor of loop formation. In silico molecular docking predicted a good binding score between Rh2 or PPD and the ATP-binding pocket of VEGFR2. VEGF bioassay showed that Rh2 was an allosteric modulator of VEGFR2. In addition, SRh2 and PPD had good binding scores with AQP1 and AQP5, both of which play roles in cell migration and proliferation. Conclusion: The combination of these molecules might be responsible for the anti-cancer effects observed by Rg3.

• BMB Reports
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• v.53 no.7
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• pp.357-366
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• 2020

Currently, most biological research relies on conventional experimental techniques that allow only static analyses at certain time points in vitro or ex vivo. However, if one could visualize cellular dynamics in living organisms, that would provide a unique opportunity to study key biological phenomena in vivo. Intravital microscopy (IVM) encompasses diverse optical systems for direct viewing of objects, including biological structures and individual cells in live animals. With the current development of devices and techniques, IVM addresses important questions in various fields of biological and biomedical sciences. In this mini-review, we provide a general introduction to IVM and examples of recent applications in the field of immunology, oncology, and vascular biology. We also introduce an advanced type of IVM, dubbed real-time IVM, equipped with video-rate resonant scanning. Since the realt-ime IVM can render cellular dynamics with high temporal resolution in vivo, it allows visualization and analysis of rapid biological processes.

• Journal of Medicine and Life Science
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• v.21 no.3
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• pp.117-120
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• 2024

Fetal ventriculomegaly (VM) is a relatively common finding during prenatal examinations and occurs in approximately 0.2% of live births. Although there are various causes, obstructive VM due to cerebellar hemorrhage is exceedingly rare. A 33-year-old primigravida presented at 32 weeks of gestation with VM. At 36 weeks of age, a male infant was delivered via cesarean section. Postnatal imaging revealed severe bilateral hydrocephalus and space-occupying lesions in the cerebellum. Initial concerns about a potential germ cell tumor were raised due to elevated alpha-fetoprotein levels in both serum and cerebrospinal fluid. An external ventricular drain was placed to manage obstructive hydrocephalus. When the baby was 1 month old, surgical exploration revealed an old blood clot without any evidence of a tumor. Histopathological examination confirmed an old hemorrhage with no malignant cells. This case underscores the diagnostic challenges in distinguishing between hemorrhages and tumors in the context of fetal VM. Despite elevated alpha-fetoprotein levels, no tumors were identified. The underlying cause of cerebellar hemorrhage remains unclear despite extensive workups. Nevertheless, this case report details multifaceted diagnostic efforts to address the rare occurrence of cerebellar hemorrhage related to fetal VM, leading to a comprehensive case presentation.

• Journal of Biosystems Engineering
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• v.39 no.3
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• pp.244-252
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• 2014

Purpose: The development of an efficient in vitro cell culture device to process various cells would represent a major milestone in biological science and engineering. However, the current conventional macro-scale in vitro cell culture platforms are limited in their capacity for detailed analysis and determination of cellular behavior in complex environments. This paper describes a microfluidic-based culture device that allows accurate control of parameters of physical cues such as pressure. Methods: A microfluidic device, as a model microbioreactor, was designed and fabricated to culture Saccharomyces cerevisiae and Chlamydomonas reinhardtii under various conditions of physical pressure stimulus. This device was compatible with live-cell imaging and allowed quantitative analysis of physical cue-induced behavior in yeast and microalgae. Results: A simple microfluidic-based in vitro cell culture device containing a cell culture channel and an air channel was developed to investigate physical pressure stress-induced behavior in yeasts and microalgae. The shapes of Saccharomyces cerevisiae and Chlamydomonas reinhardtii could be controlled under compressive stress. The lipid production by Chlamydomonas reinhardtii was significantly enhanced by compressive stress in the microfluidic device when compared to cells cultured without compressive stress. Conclusions: This microfluidic-based in vitro cell culture device can be used as a tool for quantitative analysis of cellular behavior under complex physical and chemical conditions.

• Korean Chemical Engineering Research
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• v.59 no.3
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• pp.366-372
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• 2021

Cadherin-Catenin complex is thought to play an essential role in the transmission of force at adherens junction. Due to the lack of proper tools to visualize and detect mechanical force signals, the underlying mechanism by which the cadherin-catenin complex regulates force transmission at intercellular junctions remains elusive. In this study, we visualize cadherin-mediated force transmission using an engineered α-Catenin sensor based on fluorescence resonance energy transfer. Our results reveal that α-catenin is a key force transducer in cadherin-mediated mechanotransduction at cell-cell junctions. Thus, our finding will provide important insights for studying the effects of chemical and physical signals on cell-cell communication and the relationship between physiological and pathological phenomena.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.272-281
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• 2019

Background: Diabetic sensorineural damage is a complication of the sensory neural system, resulting from long-term hyperglycemia. Red ginseng (RG) has shown efficacy for treatment of various diseases, including diabetes mellitus; however, there is little research about its benefit for treating sensorineural damage. Therefore, we aim to evaluate RG efficacy in alloxan-induced diabetic neuromast (AIDN) zebrafish. Methods: In this study, we developed and validated an AIDN zebrafish model. To assess RG effectiveness, we observed morphological changes in live neuromast zebrafish. Also, zebrafish has been observed to have an ultrastructure of hair-cell cilia under scanning electron microscopy. Thus, we recorded these physiological traits to assess hair cell function. Finally, we confirmed that RG promoted neuromast recovery via nerve growth factor signaling pathway markers. Results: First, we established an AIDN zebrafish model. Using this model, we showed via live neuromast imaging that RG fostered recovery of sensorineural damage. Damaged hair cell cilia were recovered in AIDN zebrafish. Furthermore, RG rescued damaged hair cell function through cell membrane ion balance. Conclusion: Our data suggest that RG potentially facilitates recovery in AIDN zebrafish, and its mechanism seems to be promotion of the nerve growth factor pathway through increased expression of topomyosin receptor kinase A, transient receptor potential channel vanilloid subfamily type 1, and mitogen-activated protein kinase phosphorylation.

• Ocean and Polar Research
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• v.31 no.4
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• pp.349-357
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• 2009

Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.

• Molecules and Cells
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• v.37 no.7
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• pp.526-531
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• 2014

Human PARP family consists of 17 members of which PARP-1 is a prominent member and plays a key role in DNA repair pathways. It has an N-terminal DNA-binding domain (DBD) encompassing the nuclear localisation signal (NLS), central automodification domain and C-terminal catalytic domain. PARP-1 accounts for majority of poly-(ADP-ribose) polymer synthesis that upon binding to numerous proteins including PARP itself modulates their activity. Reduced PARP-1 activity in ageing human samples and its deficiency leading to telomere shortening has been reported. Hence for cell survival, maintenance of genomic integrity and longevity presence of intact PARP-1 in the nucleus is paramount. Although localisation of full-length and truncated PARP-1 in PARP-1 proficient cells is well documented, subcellular distribution of PARP-1 fragments in the absence of endogenous PARP-1 is not known. Here we report the differential localisation of PARP-1 Nterminal fragment encompassing NLS in PARP-$1^{+/+}$ and PARP-$1^{-/-}$ mouse embryo fibroblasts by live imaging of cells transiently expressing EGFP tagged fragment. In PARP-$1^{+/+}$ cells the fragment localises to the nuclei presenting a granular pattern. Furthermore, it is densely packaged in the midsections of the nucleus. In contrast, the fragment localises exclusively to the cytoplasm in PARP-$1^{-/-}$ cells. Flourescence intensity analysis further confirmed this observation indicating that the N-terminal fragment requires endogenous PARP-1 for its nuclear transport. Our study illustrates the trafficking role of PARP-1 independently of its enzymatic activity and highlights the possibility that full-length PARP-1 may play a key role in the nuclear transport of its siblings and other molecules.