• 제목/요약/키워드: biological cells

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Bombyx mori세포주와 Spodoptera frugiperda세포주의 분자생물학적 표식자 (The Molecular Biological Marker in Bombyx mori and Spodoptera frugiperda Cells)

  • 진병래;제연호;강석권
    • 한국잠사곤충학회지
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    • 제38권1호
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    • pp.53-56
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    • 1996
  • 곤충세포주로 널리 이용되고 있는 Sf 세포주와 Bm 세포주의 분자생물학적 표식자를 탐색하기 위하여, 총 세포 단백질의 SDS-PAGE와 genomic DNA를 RAPD(Random Amplification of Polymorphic DNA) 방법으로 분석하였다. 그 결과, 총 세포 단백질 및 genomic DNA, 패턴에서 두 세포주를 뚜렷하게 구별할 수 있는 밴드를 탐색하였으며, 이들은 유용한 분자 생물학적 표식자로 이용될 수 있을 것이다.

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허혈-재관류 유도 신경세포사멸에 대한 초석잠 추출물의 신경보호 효과 연구 (Neuroprotective Effects of Stachys sieboldii Miq. Extract Against Ischemia/reperfusion-induced Apoptosis in SH-SY5Y Neuroblastoma Cells)

  • 이영경;김철환;신수영;황병수;서민정;황혜진;최경민;정진우
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2020년도 춘계학술대회
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    • pp.76-76
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    • 2020
  • Stachys sieboldii Miq. (chinese artichoke), which has been extensively used in oriental traditional medicine to treat of ischemic stroke; however, the role of Stachys sieboldii Miq. (SSM) in cerebral ischemia/reperfusion (I/R) injury is not yet fully understood. In the current study, the neuroblastoma cell line (SH-SY5Y) were subjected to oxygen-glucose deprivation/reperfusion (OGD/R) to simulate I/R injury in vitro model. The results showed that SSM improved OGD/R-induced inhibitory effect on cell viability of SH-SY5Y Cells. SSM displayed anti-oxidative activity as proved by the decreased levels of reactive oxygen species (ROS) and malondialdehyde (MDA), and increased activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in OGD/R-induced SH-SY5Y Cells. In addition, cell apoptosis was markedly decreased after SSM treatment in OGD/R-induced SH-SY5Y Cells. The up-regulation of Bcl-2 and down-regulation of Bax, thus reducing the Bax/Bcl-2 ratio that in turn protected the activation of caspase-9 and -3, and inhibition of poly (ADP-ribose) polymerase cleavage, which was associated with the blocking of cytochrome c release to the cytoplasm. Collectively, SSM protected human neuroblastoma SH-SY5Y cells from OGD/R-induced injury via preventing mitochondrial-dependent pathway through scavenging excessive ROS, suggesting that SSM might be a potential agent for the ischemic stroke therapy.

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Anticancer activity of gomisin J from Schisandra chinensis fruit

  • Samil Jung;Hyung-In Moon;Subeen Kim;Nguyen Thi Ngoc Quynh;Jimin Yu;Zolzaya Sandag;Dan-Diem Thi Le;Hyegyeong Lee;Hyojeong Lee;Myeong-Sok Lee
    • Oncology Letters
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    • 제41권1호
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    • pp.711-717
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    • 2019
  • In attempting to identify effective anticancer drugs from natural products that are harmless to humans, we found that the gomisin J from Schisandra chinensis fruit has anticancer activity. Schisandra chinensis fruits are used in traditional herbal medicine and gomisin J is one of their chemical constituents. In the present study, we examined the anticancer activity of gomisin J in MCF7 and MDA-MB-231 breast cancer cell lines and in MCF10A normal cell line, in a time- and concentration-dependent manner. Our data revealed that gomisin J exerted a much stronger cytotoxic effect on MCF7 and MDA-MB-231 cancer cells than on MCF10A normal cells. Gomisin J suppressed the proliferation and decreased the viability of MCF7 and MDA-MB-231 cells at relatively low (<10 ㎍/ml) and high (>30 ㎍/ml) concentrations, respectively. Our data also revealed that gomisin J induced necroptosis, a programmed form of necrosis, as well as apoptosis. Notably, gomisin J predominantly induced necroptosis in MCF7 cells that are known to have high resistance to many pro-apoptotic anticancer drugs, while MDA-MB-231 exhibited a much lower level of necroptosis but instead a higher level of apoptosis. This data indicated the possibility that it may be used as a more effective anticancer drug, especially in apoptosis-resistant malignant cancer cells. In an extended study, gomisin J exhibited a strong cytotoxic effect on all tested various types of 13 cancer cell lines, indicating its potential to be used against a wide range of different types of cancer cells.

Enhanced Sialylation of Recombinant Erythropoietin in CHO Cells by Human Glycosyltransferase Expression

  • Jeong, Yeon-Tae;Choi, One;Lim, Hye-Rim;Son, Young-Dok;Kim, Hong-Jin;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • 제18권12호
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    • pp.1945-1952
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    • 2008
  • Sialylation, the attachment of sialic acid residues to a protein, can affect the biological activity and in vivo circulatory half-life of glycoproteins. Human ${\alpha}2$,3-sialyltransferase (${\alpha}2$,3-ST) and ${\beta}1$,4-galactosyltransferase (${\beta}1$,4-GT) are responsible for terminal sialylation and galactosylation, respectively. Enhanced sialylation of human erythropoietin (EPO) by the expression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT was achieved using recombinant Chinese hamster ovary (CHO) cells (EC1). The sialic acid content and sialylation of N-glycans were evaluated by HPLC. When ${\alpha}2$,3-ST was expressed in CHO cells (EC1-ST2), the sialic acid content (moles of sialic acid/mole of EPO) increased from 6.7 to 7.5. In addition, the amount of trisialylated glycans increased from 17.3% to 26.1 %. When ${\alpha}2$,3-ST and ${\beta}1$,4-GT were coexpressed in CHO cells (EC1-GTST15), the degree of sialylation was greater than that in EC1-ST2 cells. In the case of EC1-GTST15 cells, the sialic acid content increased to 8.2 and the proportion of trisialylated glycans was markedly increased from 17.3% to 35.5%. Interestingly, the amount of asialoglycans decreased only in the case of GTST15 cells (21.4% to 14.2%). These results show that coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT is more effective than the expression of ${\alpha}2$,3-ST alone. Coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT did not affect CHO cell growth and metabolism or EPO production. Thus, coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT may be beneficial for producing therapeutic glycoproteins with enhanced sialylation in CHO cells.

G-Protein-Coupled Receptor 120 Mediates DHA-Induced Apoptosis by Regulating IP3R, ROS and, ER Stress Levels in Cisplatin-Resistant Cancer Cells

  • Shin, Jong-Il;Jeon, Yong-Joon;Lee, Sol;Lee, Yoon Gyeong;Kim, Ji Beom;Lee, Kyungho
    • Molecules and Cells
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    • 제42권3호
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    • pp.252-261
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    • 2019
  • The omega-3 fatty acid docosahexaenoic acid (DHA) is known to induce apoptosis and cell cycle arrest via the induction of reactive oxygen species (ROS) production and endoplasmic reticulum (ER) stress in many types of cancers. However, the roles of DHA in drug-resistant cancer cells have not been elucidated. In this study, we investigated the effects of DHA in cisplatin-resistant gastric cancer SNU-601/cis2 cells. DHA was found to induce ROS-dependent apoptosis in these cells. The inositol 1,4,5-triphosphate receptor ($IP_3R$) blocker 2-aminoethyl diphenylboninate (2-APB) reduced DHA-induced ROS production, consequently reducing apoptosis. We also found that G-protein-coupled receptor 120 (GPR120), a receptor of long-chain fatty acids, is expressed in SNU-601/cis2 cells, and the knockdown of GPR120 using specific shRNAs alleviated DHA-mediated ROS production and apoptosis. GPR120 knockdown reduced the expression of ER stress response genes, similar to the case for the pre-treatment of the cells with N-acetyl-L-cysteine (NAC), an ROS scavenger, or 2-APB. Indeed, the knockdown of C/EBP homologous protein (CHOP), a transcription factor that functions under ER stress conditions, markedly reduced DHA-mediated apoptosis, indicating that CHOP plays an essential role in the anti-cancer activity of DHA. These results suggest that GPR120 mediates DHA-induced apoptosis by regulating $IP_3R$, ROS, and ER stress levels in cisplatin-resistant cancer cells, and that GPR120 is an effective chemotherapeutic target for cisplatin resistance.

An Antisense Oligodeoxynucleotide to the LH Receptor Attenuates FSH-induced Oocyte Maturation in Mice

  • Yang, Jiange;Fu, Maoyong;Wang, Songbo;Chen, Xiufen;Ning, Gang;Xu, Baoshan;Ma, Yuzhen;Zhang, Meijia;Xia, Guoliang
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권7호
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    • pp.972-979
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    • 2008
  • It has been recently shown that expression of the LH receptor (LHR) in cumulus cells is related with FSH-induced meiotic resumption of mouse cumulus enclosed oocytes (CEOs). However, to date, it is still unclear whether LHR expression in cumulus cells plays a key role during FSH-induced oocyte maturation. The purpose of this study was to characterize the functional role of LHRs in cumulus cells. CEOs were isolated from eCG-primed preovulatory follicles and cultured in hypoxanthine (HX) arrested medium. LHR protein expression in cumulus cells was time-dependent increasing during the process of FSH-induced oocyte maturation. While the sense oligodeoxynucleotide (ODN) had no effect, antisense ODN inhibited FSH-induced LHR expression and meiotic resumption. Moreover, this antisense ODN against LHR could inhibit FSH-induced mitogen-activated protein kinase (MAPK) phosphorylation. This study suggested that LHR expression in cumulus cells is involved in FSH-induced oocyte meiotic resumption, which process is possibly regulated by MAPK cascade.

A Systematic Analysis of Drosophila Regulatory Peptide Expression in Enteroendocrine Cells

  • Chen, Ji;Kim, Seol-min;Kwon, Jae Young
    • Molecules and Cells
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    • 제39권4호
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    • pp.358-366
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    • 2016
  • The digestive system is gaining interest as a major regulator of various functions including immune defense, nutrient accumulation, and regulation of feeding behavior, aside from its conventional function as a digestive organ. The Drosophila midgut epithelium is completely renewed every 1-2 weeks due to differentiation of pluripotent intestinal stem cells in the midgut. Intestinal stem cells constantly divide and differentiate into enterocytes that secrete digestive enzymes and absorb nutrients, or enteroendocrine cells that secrete regulatory peptides. Regulatory peptides have important roles in development and metabolism, but study has mainly focused on expression and functions in the nervous system, and not much is known about the roles in endocrine functions of enteroendocrine cells. We systemically examined the expression of 45 regulatory peptide genes in the Drosophila midgut, and verified that at least 10 genes are expressed in the midgut enteroendocrine cells through RT-PCR, in situ hybridization, antisera, and 25 regulatory peptide-GAL transgenes. The Drosophila midgut is highly compartmentalized, and individual peptides in enteroendocrine cells were observed to express in specific regions of the midgut. We also confirmed that some peptides expressed in the same region of the midgut are expressed in mutually exclusive enteroendocrine cells. These results indicate that the midgut enteroendocrine cells are functionally differentiated into different subgroups. Through this study, we have established a basis to study regulatory peptide functions in enteroendocrine cells as well as the complex organization of enteroendocrine cells in the Drosophila midgut.

감마선에 조사된 생쥐에 있어서 방사선방어효과 평가를 위한 생물학적 파라메타 (Biological Parameters for Assessing Radioprotective Effects in ${\gamma}-irradiated$ Mice)

  • 천기정;김봉희;이영근;김진규
    • 약학회지
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    • 제43권2호
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    • pp.278-284
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    • 1999
  • This study deals with the biological changes in mice after ${\gamma}-irradiated$. Four weeks old BALA/c mice were irradiated with 6.5Gy of ${\gamma}-ray$ on the fifth day after oral administration of radioprotectants such as ascorbic acid, tocopherol and cysteine. Control group was irradiated with 6.5Gy without pre-administration of radioprotectors. Blood cells and sperm cells were counted and body, testis and spleen were weighed 3 days after irradiation. And also liver antioxidant activity and range of spleen immune cells were measured. Differences in most biological parameters were not clearly distinguished between experimental groups. However, the relative spleen weight, the relative testis weight and the population size of spleen immune cells such as T helper cells, B cells and macrophages measured by means of FACS showed significant difference between irradiated and radioprotectant administered group. It is concluded that the relative spleen weight, the relative testis weight and the population size of spleen immune cells are easy and useful parameters for assessing the effect of radioprotective substances and for quantifying biological damage of radiation, as well.

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Anti-Proliferative Activities of Vasicinone on Lung Carcinoma Cells Mediated via Activation of Both Mitochondria-Dependent and Independent Pathways

  • Dey, Tapan;Dutta, Prachurjya;Manna, Prasenjit;Kalita, Jatin;Boruah, Hari Prasanna Deka;Buragohain, Alak Kumar;Unni, Balagopalan
    • Biomolecules & Therapeutics
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    • 제26권4호
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    • pp.409-416
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    • 2018
  • Vasicinone, a quinazoline alkaloid from Adhatoda vasica Nees. is well known for its bronchodilator activity. However its anti-proliferative activities is yet to be elucidated. Here-in we investigated the anti-proliferative effect of vasicinone and its underlying mechanism against A549 lung carcinoma cells. The A549 cells upon treatment with various doses of vasicinone (10, 30, 50, $70{\mu}M$) for 72 h showed significant decrease in cell viability. Vasicinone treatment also showed DNA fragmentation, LDH leakage, and disruption of mitochondrial potential, and lower wound healing ability in A549 cells. The Annexin V/PI staining showed disrupted plasma membrane integrity and permeability of PI in treated cells. Moreover vasicinone treatment also lead to down regulation of Bcl-2, Fas death receptor and up regulation of PARP, BAD and cytochrome c, suggesting the anti-proliferative nature of vasicinone which mediated apoptosis through both Fas death receptors as well as Bcl-2 regulated signaling. Furthermore, our preliminary studies with vasicinone treatment also showed to lower the ROS levels in A549 cells and have potential free radical scavenging (DPPH, Hydroxyl) activity and ferric reducing power in cell free systems. Thus combining all, vasicinone may be used to develop a new therapeutic agent against oxidative stress induced lung cancer.