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Characterization of intracellular Ca2+ mobilization in gefitinib-resistant oral squamous carcinoma cells HSC-3 and -4

  • Kim, Mi Seong;Kim, Min Seuk
    • International Journal of Oral Biology
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    • v.46 no.4
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    • pp.176-183
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    • 2021
  • Oral squamous cell carcinoma (OSCC) metastasis is characterized by distant metastasis and local recurrence. Combined chemotherapy with cisplatin and 5-fluorouracil is routinely used to treat patients with OSCC, and the combined use of gefitinib with cytotoxic drugs has been reported to enhance the sensitivity of cancer cells in vitro. However, the development of drug resistance because of prolonged chemotherapy is inevitable, leading to a poor prognosis. Therefore, understanding alterations in signaling pathways and gene expression is crucial for overcoming the development of drug resistance. However, the altered characterization of Ca2+ signaling in drug-resistant OSCC cells remains unclear. In this study, we investigated alterations in intracellular Ca2+ ([Ca2+]i) mobilization upon the development of gefitinib resistance in human tongue squamous carcinoma cell line (HSC)-3 and HSC-4 using ratiometric analysis. This study demonstrated the presence of altered epidermal growth factor- and purinergic agonist-mediated [Ca2+]i mobilization in gefitinib-resistant OSCC cells. Moreover, Ca2+ content in the endoplasmic reticulum, store-operated calcium entry, and lysosomal Ca2+ release through the transient receptor potential mucolipin 1, were confirmed to be significantly reduced upon the development of apoptosis resistance. Consistent with [Ca2+]i mobilization, we identified modified expression levels of Ca2+ signaling-related genes in gefitinib-resistant cells. Taken together, we propose that the regulation of [Ca2+]i mobilization and related gene expression can be a new strategy to overcome drug resistance in patients with cancer.

Expression of Sodium/iodide Symporter Transgene in Neural Stem Cells (신경줄기세포(HB1.F3)에서 나트륨옥소 공동수송체 도입유전자 발현)

  • Kim, Yun-Hui;Lee, Dong-Soo;Kang, Joo-Hyun;Lee, Yong-Jin;Chung, June-Key;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.1
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    • pp.99-108
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    • 2004
  • Purpose: The ability to noninvasively track the migration of neural progenitor cells would have significant clinical and research implications. We generated stably transfected F3 human neural progenitor cells with human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation. Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent (5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to $20{\mu}M$, up to 62.5nM, respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell passage. Therefore, the gene ex[ressopm of exogenous gene transferred human stem cell might be affected to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture conditions.

Micromorphological and Chemical Characteristics of Cengal (Neobalanocarpus heimii) Heartwood Decayed by Soft Rot Fungi

  • Kim, Yoon Soo;Singh, Adya P.;Wong, Andrew H.H.;Eom, Tae-Jin;Lee, Kwang Ho
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.2
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    • pp.68-77
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    • 2006
  • The heartwood of cengal (Neobalanocarpus heimii) is known to have a high degree of decay resistance by virtue of its high extractive content. After 30 years in ground contact an utility pole of this tropical hardwood was found to be degraded only in the surface layers by cavity-forming soft rot fungi. The present work was undertaken 1) to characterize the degradation of cengal heartwood from the aspect of ultrastructure and chemistry and 2) to investigate the correlation between soft rot decay and its extractive microdistribution in wood tissues. The chemical analysis of cengal heartwood revealed the presence of a high amount of extractives as well as lignin. The wood contained a relatively high amount of condensed lignin and the guaiacyl units. Microscopic observations revealed that vessels, fibers and parenchyma cells (both ray and axial parenchyma) all contained extractives in their lumina, but in variable amounts. The lumina of fibers and most axial parenchyma were completely or almost completely filled with the extractives. TEM micrographs showed that cell walls were also impregnated with extractives and that pit membranes connecting parenchyma cells were well coated and impregnated with extractives. However, fungal hyphae were present in the extractive masses localized in cell lumina, and indications were that the extractives did not completely inhibit fungal growth. The extent of cell wall degradation varied with tissue types. The fibers appeared to be more susceptible to decay than vessels and parenchyma. Middle lamella was the only cell wall region which remained intact in all cell types which were severely degraded. The microscopic observations suggested a close correlation between extractive microdistribution and the pattern and extent of cell wall degradation. In addition to the toxicity to fungi, the physical constraint of the extractive material present in cengal heartwood cells is likely to have a profound effect on the growth and path of invasion of colonizing fungi, thus conferring protection to wood by restricting fungal entry into cell walls. The presence of relatively high amount of condensed lignin is also likely to be a factor in the resistance of cengal heartwood to soft rot decay.

Propofol protects human keratinocytes from oxidative stress via autophagy expression

  • Yoon, Ji-Young;Jeon, Hyun-Ook;Kim, Eun-Jung;Kim, Cheul-Hong;Yoon, Ji-Uk;Park, Bong-Soo;Yu, Su-Bin;Kwak, Jin-Won
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.17 no.1
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    • pp.21-28
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    • 2017
  • Background: The skin consists of tightly connected keratinocytes, and prevents extensive water loss while simultaneously protecting against the entry of microbial pathogens. Excessive cellular levels of reactive oxygen species can induce cell apoptosis and also damage skin integrity. Propofol (2,6-diisopropylphenol) has antioxidant properties. In this study, we investigated how propofol influences intracellular autophagy and apoptotic cell death induced by oxidative stress in human keratinocytes. Method: The following groups were used for experimentation: control, cells were incubated under normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$) without propofol; hydrogen peroxide ($H_2O_2$), cells were exposed to $H_2O_2$ ($300{\mu}M$) for 2 h; propofol preconditioning (PPC)/$H_2O_2$, cells pretreated with propofol ($100{\mu}M$) for 2 h were exposed to $H_2O_2$; and 3-methyladenine $(3-MA)/PPC/H_2O_2$, cells pretreated with 3-MA (1 mM) for 1 h and propofol were exposed to $H_2O_2$. Cell viability, apoptosis, and migration capability were evaluated. Relation to autophagy was detected by western blot analysis. Results: Cell viability decreased significantly in the $H_2O_2$ group compared to that in the control group and was improved by propofol preconditioning. Propofol preconditioning effectively decreased $H_2O_2$-induced cell apoptosis and increased cell migration. However, pretreatment with 3-MA inhibited the protective effect of propofol on cell apoptosis. Autophagy was activated in the $PPC/H_2O_2$ group compared to that in the $H_2O_2$ group as demonstrated by western blot analysis and autophagosome staining. Conclusion: The results suggest that propofol preconditioning induces an endogenous cellular protective effect in human keratinocytes against oxidative stress through the activation of signaling pathways related to autophagy.

Cell cycle-dependent entry of Toxoplasma gondii into synchronized HL-60 cells (세포 주기 변화에 따른 Toxoplasmu gondii의 침투 양상)

  • 윤지혜;남호우
    • Parasites, Hosts and Diseases
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    • v.29 no.2
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    • pp.121-128
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    • 1991
  • The degree of attraction of Toxoplasma gondii to vertebrate cells varies with cell type and cell phase. Human promyelocytic leukemia cells, HL-60, were synchronized by double thymidine block method and co-cultured with Toxoplasma for 1 hr at each cell stage to investigate the cell cycle specific susceptibility of parasites to host cells. For 30 hr the average number of Texoplasma that invaded was a little changed except at 3 hr from G1/S phase boundary which concurred with the peak point of DNA synthesis. At 3 hr which is a relatively short interval compared to whole S phase, modification of cells by parasitic invasion was most remarkable. The number of Toxoplasma that penetrated was increased to more than sin times. The shape of the cells became sludgy and almost indiscernible by strong accessibility of parasites only for an hour of mfd-S phase. The same auctuation was also observed at the second peak of S phase but weakly. This suggests that there be surface molecules concerning with the attachment of Texoplasma to the host cells, which is expressed at special point of S phase. further studies on the specific protein or similar molecules related could be carried out using synchronized HL-60 cells.

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Substrate roughness induces the development of defective E-cadherin junctions in human gingival keratinocytes

  • Jin, Chengbiao;Lee, Gayoung;Oh, Changseok;Kim, Hyun Jung;Kim, Hyun-Man
    • Journal of Periodontal and Implant Science
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    • v.47 no.2
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    • pp.116-131
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    • 2017
  • Purpose: The entry of bacteria or harmful substances through the epithelial seal of human gingival keratinocytes (HGKs) in the junctional epithelium (JE) is blocked by specialized intercellular junctions such as E-cadherin junctions (ECJs). However, the influence of roughened substrates, which may occur due to apical migration of the JE, root planing, or peri-implantitis, on the development of the ECJs of HGKs remains largely unknown. Methods: HGKs were cultured on substrates with varying levels of roughness, which were prepared by rubbing hydrophobic polystyrene dishes with silicon carbide papers. The activity of c-Jun N-terminal kinase (JNK) was inhibited with SP600125 or by transfection with JNK short hairpin RNA. The development of intercellular junctions was analyzed using scanning electron microscopy or confocal laser scanning microscopy after immunohistochemical staining of the cells for E-cadherin. The expression level of phospho-JNK was assessed by immunoblotting. Results: HGKs developed tight intercellular junctions devoid of wide intercellular gaps on smooth substrates and on rough substrates with low-nanometer dimensions (average roughness $[Ra]=121.3{\pm}13.4nm$), although the ECJs of HGKs on rough substrates with low-nanometer dimensions developed later than those of HGKs on smooth substrates. In contrast, HGKs developed short intercellular junctions with wide intercellular gaps on rough substrates with mid- or high-nanometer dimensions ($Ra=505.3{\pm}115.3nm$, $867.0{\pm}168.6nm$). Notably, the stability of the ECJs was low on the rough substrates, as demonstrated by the rapid destruction of the cell junction following calcium depletion. Inhibition of JNK activity promoted ECJ development in HGKs. JNK was closely associated with cortical actin in the regulation of ECJs in HGKs. Conclusions: These results indicate that on rough substrates with nanometer dimensions, the ECJs of HGKs develop slowly or defectively, and that this effect can be reversed by inhibiting JNK.

Technology Development of Entry-Level MiC Smart Photovoltaic System based on SOC (SoC 기반 보급형 MiC 스마트 태양광발전시스템 기술개발)

  • Yoon, Yongho
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.20 no.3
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    • pp.129-134
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    • 2020
  • Moisture infiltration inside the solar cell module, filling of EVA sheet, melting of the frame seal, and deterioration of power generation performance in the module one year after installation are occurring. Whitening phenomenon, electrode corrosion phenomenon, and dielectric breakdown phenomenon are appearing in solar cell module installed in Korea before 5-7 years, leading to deterioration of power generation performance, and big problems for long-term reliability and long life technology are emerging. Therefore, in order to solve these problems, the development of a micro inverter (MiCrco Inverter Converter, MiC) including the function of securing the durability of the solar cell module and monitoring the aging progress and the solar cell based on the monitoring data from the MiC smart monitoring programs have been proposed to determine the aging of modules. In addition, in order to become a highly efficient solar smart monitoring system through systematic operation management through IT convergence with MiC that has enhanced monitoring function of solar cell module, SoC(System On Chip) in micro inverter is the environment for solar cell module. There is a demand for functions that can detect information in a complex manner and perform communication and control when necessary. Based on these requirements, this paper aims to develop SoC-based low-cost MiC smart photovoltaic system technology.

Brief Review of Silicon Solar Cells (실리콘 태양전지)

  • Yi, Jun-Sin
    • Journal of the Korean Vacuum Society
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    • v.16 no.3
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    • pp.161-166
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    • 2007
  • Photovoltaic (PV) technology permits the transformation of solar light directly into electricity. For the last five years, the photovoltaic sector has experienced one of the highest growth rates worldwide (over 30% in 2006) and for the next 20 years, the average production growth rate is estimated to be between 27% and 34% annually. Currently the cost of electricity produced using photovoltaic technology is above that for traditional energy sources, but this is expected to fall with technological progress and more efficient production processes. A large scale production of solar grade silicon material of high purity could supply the world demand at a reasonably lower cost. A shift from crystalline silicon to thin film is expected in the future. The technical limit for the conversion efficiency is about 30%. It is assumed that in 2030 thin films will have a major market share (90%) and the share of crystalline cells will have decreased to 10%. Our research at Sungkyunkwan University of South Korea is confined to crystalline silicon solar cell technology. We aim to develop a technology for low cost production of high efficiency silicon solar cell. We have successfully fabricated silicon solar cells of efficiency more than 16% starting with multicrystalline wafers and that of efficiency more than 17% on single crystalline wafers with screen printing metallization. The process of transformation from the first generation to second generation solar cell should be geared up with the entry of new approaches but still silicon seems to remain as the major material for solar cells for many years to come. Local barriers to the implementation of this technology may also keep continuing up to year 2010 and by that time the cost of the solar cell generated power is expected to be 60 cent per watt. Photovoltaic source could establish itself as a clean and sustainable energy alternate to the ever depleting and polluting non-renewable energy resource.

Essential role of tryptophan residues in toxicity of binary toxin from Bacillus sphaericus

  • Kunthic, Thittaya;Promdonkoy, Boonhiang;Srikhirin, Toemsak;Boonserm, Panadda
    • BMB Reports
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    • v.44 no.10
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    • pp.674-679
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    • 2011
  • Bacillus sphaericus produces mosquito-larvicidal binary toxin composed of BinA and BinB. While BinB is expected to bind to a specific receptor on the cell membrane, BinA interacts to BinB or BinB receptor complex and translocates into the cytosol to exert its activity via unknown mechanism. To investigate functional roles of aromatic cluster in BinA, amino acids at positions Y213, Y214, Y215, W222 and W226 were substituted by leucine. All mutant proteins were highly produced and their secondary structures were not affected by these substitutions. All mutants are able to insert into lipid monolayers as observed by Langmuir-Blodgett trough and could permeabilize the liposomes in a similar manner as the wild type. However, mosquito-larvicidal activity was abolished for W222L and W226L mutants suggesting that tryptophan residues at both positions play an important role in the toxicity of BinA, possibly involved in the cytopathological process after toxin entry into the cells.

Use of Androgenesis in Haploid Breeding

  • Yi, Gihwan;Kim, Kyung-Min;Sohn, Jae-Keun
    • Current Research on Agriculture and Life Sciences
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    • v.31 no.2
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    • pp.75-82
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    • 2013
  • Haploids are plants with a gametophytic number of chromosomes in their sporophytes. Androgenesis occurs from asymmetric division of pollen grains into generative cells and vegetative cells, followed by re-entry of the vegetative cell during S-phase, which causes microspores progress into G2/M transition in culture. One of the most interesting features of haploids is the possibility to produce doubled haploid (DH) individuals. Doubled haploidy is extremely useful to plant breeders because it enables shortened breeding periods and efficiency in selection of useful recessive agronomic traits. Doubled-haploid technology is not only applicable to breeding, but also to transformation programs of desired genes. In addition to practical breeding programs, DH lines provide useful materials of fundamental genetics including exploitation of QTLs and genes conferred with various agronomic traits by establishing DH populations. This paper provides historical overviews on androgenesis and describes several mechanisms associated with pollen embryogenesis, including mode of actions in pollen embryogenesis, mechanisms of chromosome doubling and factors affecting androgenesis. We also discuss recent progress in application of haploids to breeding, genes associated with in vitro response and drawbacks to anther culture for application of doubled haploids in crop breeding.

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