• Journal of Life Science
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• v.26 no.8
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• pp.875-880
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• 2016

The plasma membrane plays a crucial role in relaying signals from the outside environment to the inside of the cells. In eukaryotic cells, the inner leaflets of the plasma membrane are composed mostly of phospholipids, including phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositides (PIs). In this study, we tried to analyze the morphological changes induced by EGFP-fused membrane binding proteins, which are targeted to the plasma membrane via specific phospholipids binding. As a result, we found that overexpression of EGFP-P4M-SidM, a specific PI4P binding protein, or EGFP alone, did not induce any morphological changes. On the other hand, overexpression of EGFP-PLCδ1(PH), which is a specific PI(4,5)P₂ binding protein, EGFP-AKT1(PH) which binds to PI(3,4,5)P₃, or EGFP-OSH2(PH)×2 which binds to PI4P and PI(4,5)P₂, could induce the filopodia and lamilapodia formation as well as cell shrinkage. Overexpression of Lact-C2-EGFP which is a specific PS-binding probe, EGFP fused Aplysia phosphodiesterase 4 (ApPDE4) long-form (L(N20)-EGFP) which is localized to the plasma membrane via hydrophobic interaction, or EGFP fused Aplysia PDE4 short-form (S(N-UCR1-2)-EGFP) which is localized to the plasma membrane via electrostatic interaction, could induce cell shrinkage, but not filopodia or lamilapodia formation. Taken together, our data support that the different phospholipid bindings in the plasma membrane could induce different characteristic morphological changes. Thus, we can analyze, characterize, and classify the cellular morphological changes induced by the various phospholipid binding proteins.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.9
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• pp.1127-1132
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• 2006

In our previous study, a novel herb mixture (HIM-I) of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. A new herbal composition (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. HemoHIM showed more effective than HIM-I in immune modulation as well as radioprotection. The present study is designed to investigate the protective effects of HIM-I, HIM-I-P, and HemoHIM on hydrogen peroxide $(H_2O_2)$ induced apoptosis of human promyelocytic leukemia (HL-60) cells. It was shown that $H_2O_2$ treatment reduced the viability of cells, and increased appearance of DNA ladders, hypodiploid (subG1) cells, and phosphatidylserine translocation level. Pretreatment of HemoHIM significantly reduced the cytotoxic effect induced by $H_2O_2$, associated with reducing the translocation of phosphatidylserine, hypodiploid cells and DNA ladders. HemoHIM appeared to be more protective than HIM-I against $H_2O_2$ induced apoptosis whereas, it exhibited similar activity to HIM-I-P. These results indicated that HemoHIM might be an useful agent for protection against oxidative stress $(H_2O_2)-induced$ apoptosis as well as immune modulation, especially since it is a relatively nontoxic natural product.

• The Journal of Internal Korean Medicine
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• v.21 no.2
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• pp.259-266
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• 2000

Objectives : The effects of aqueous extracts of Cortex ulmi pumilae (a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origm hepatoma cell lines, HepG2. Methods : The death of HepG2 cells was markedly induced by the addition of extracts of Cortex ulmi pumilae in a dose-dependent manner. The apoptotic characteristic ladder pattern of DNA strand break was not observed in cell death of HepG2. In addition, it was not shown nucleus chromatin condensation and fragmentation under hoechst staining. However, by the using annexin V staining assay, externalizations of phosphatidylserine in HepG2 cell which were treated with Cortex ulmi pumilae extracts were detected in the early time (at 9 hr after extract treatment). Furthermore, LDH release was not detected in this early stage. Therefore, Cortex ulmi pumilae extracts-induced cell death of HepG2 cells is mediated by apoptotic death signal processes. Result : The activity of caspase 3-like proteases remained in a basal level in HepG2 cells which treated with the extract of Cordyceps sinensis. However, it was markedly increased in HepG2 cells which treated with two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) which were differently extracted (respectively, 2.3 and 3.3 fold). On a while, the phosphotransferase activities of JNK1 was markedly induced in HepG2 cells which were treated with two extracts of Cortex ulmi pumilae. On the contrary, the activation of transcriptional activator, activating protein1(AP-1) and NF-kB were severely decreased by these two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K). In addition, antioxidants (GSH and NAC) and intracellular $Ca2^+$ level regulator (Bapta/AM and Thapsigargin) did not affect Cortex ulmi pumilae extracts-induced apoptotic death of HepG2 cells. Conclusions : In conclusion, our results suggest that two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) induces the apoptotic death of human liver origin hepatoma HepG2 cells via activation of caspase 3-like proteases as well as JNK1, and inhibition of transcriptional activators, AP-1 and $NK-{\kappa}B$.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.197-205
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• 2017

Exposure of Jurkat T cell clone (J/Neo cells) to acacetin (5,7-dihydroxy-4'-methoxyflavone), which is present in barnyard millet (Echinochloa esculenta (A. Braun)) grains, caused cytotoxicity, enhancement of apoptotic $sub-G_1$ rate, Bak activation, loss of mitochondrial membrane potential (${\Delta}{\Psi}m$), activation of caspase-9 and caspase-3, degradation of poly(ADP-ribose) polymerase, and FITC-Annexin V-stainable phosphatidylserine exposure on the external surface of the cytoplasmic membrane without accompanying necrosis. These apoptotic responses were abrogated in Jurkat T cell clone (J/Bcl-xL) overexpressing Bcl-xL. Under the same conditions, cellular autophagic responses, including suppression of the Akt-mTOR pathway and p62/SQSTM1 down-regulation, were commonly detected in J/Neo and J/Bcl-xL cells; however, formation of acridine orange-stainable acidic vascular organelles, LC3-I/II conversion, and Beclin-1 phosphorylation (Ser-15) were detected only in J/Neo cells. Correspondingly, concomitant treatment with the autophagy inhibitor (3-methyladenine or LY294002) appeared to enhance acacetin-induced apoptotic responses, such as Bak activation, ${\Delta}{\Psi}m$ loss, activation of caspase-9 and caspase-3, and apoptotic $sub-G_1$ accumulation. This indicated that acacetin could induce apoptosis and cytoprotective autophagy in Jurkat T cells simultaneously. Together, these results demonstrate that acacetin induces not only apoptotic cell death via activation of Bak, loss of ${\Delta}{\Psi}m$, and activation of the mitochondrial caspase cascade, but also cytoprotective autophagy resulting from suppression of the Akt-mTOR pathway. Furthermore, pharmacologic inhibition of the autophagy pathway augments the activation of Bak and resultant mitochondrial damage-mediated apoptosis in Jurkat T cells.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8503-8512
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• 2016

The purpose of this study was to provide a detailed explanation of the mechanism of bisanthracycline, WP 631 in comparison to doxorubicin (DOX), a first generation anthracycline, currently the most widely used pharmaceutical in clinical oncology. Experiments were performed in SKOV-3 ovarian cancer cells which are otherwise resistant to standard drugs such as cis-platinum and adriamycin. As attention was focused on the ability of WP 631 to induce apoptosis, this was examined using a double staining method with Annexin V and propidium iodide probes, with measurement of the level of intracellular calcium ions and cytosolic cytochrome c. The western blotting technique was performed to confirm PARP cleavage. We also investigated the involvement of caspase activation and DNA degradation (comet assay and immunocytochemical detection of phosphorylated H2AX histones) in the development of apoptotic events. WP 631 demonstrated significantly higher effectiveness as a pro-apoptotic drug than DOX. This was evident in the higher levels of markers of apoptosis, such as the externalization of phosphatidylserine and the elevated level of cytochrome c. An extension of incubation time led to an increase in intracellular calcium levels after treatment with DOX. Lower changes in the calcium content were associated with the influence of WP 631. DOX led to the activation of all tested caspases, 8, 9 and 3, whereas WP 631 only induced an increase in caspase 8 activity after 24h of treatment and consequently led to the cleavage of PARP. The lack of active caspase 3 had no outcome on the single and double-stranded DNA breaks. The obtained results show that WP 631 was considerably more genotoxic towards the investigated cell line than DOX. This effect was especially visible after longer times of incubation. The above detailed studies indicate that WP 631 generates early apoptosis and cell death independent of caspase-3, detected at relatively late time points. The observed differences in the mechanisms of the action of WP631 and DOX suggest that this bisanthracycline can be an effective alternative in ovarian cancer treatment.

• Korean Journal of Food Science and Technology
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• v.26 no.2
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• pp.123-126
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• 1994

Changes in lipid components of pollack meat during sun-drying and effects of NaCl on lipid oxidation were examined. TBA values and peroxide values of sun dried pollack(SD), salted and sun dried pollack (SS) were 0.142 and 14.8 meq/kg, 0.226 and 20.0 meq/kg after sun-drying, respectively. Raw pollack contained 6.12% total lipid consisted of 2.42% neutral lipid(NL) and 3.70% phospholipid(PL) as dry basis, and there were $47{\sim}65%$ decrease in PL content during sun-drying. The NL class of raw pollack mainly consisted of triglyceride(TG), sterol(ST)+diglyceride(DG), hydrocarbon(HC)+sterol ester(SE), and main components in PL class were phosphatidylcholine(PC), phosphatidylethanolamlne(PE) and phosphatidylserine(PS). The contents of TG, ST+DG, PC and PE decreased, while those of free fatty acid, HC+SE, sphingomyelin and lysophosphatidylcholine increased markedly during sun-drying. The major fatty acids of TL in raw pollack, PD and SD samples were generally 22:6, 16:0, 20:5, 18:1 and 18:3; 20:5 decreased markedly during sun-drying, while saturates and monoenes such as 16:0, 18:0 and 18:1 increased slightly. And remaining ratios of polyunsaturated fatty acids of TL, NL and PL in SD and SS samples were 81.1%, 92.5%. 73.3%, and 74.6%, 74.1%, 45.4%, respectively. The results of changes in lipid components during sun-drying showed that sodium chloride catalyzed the lipid oxidation of pollack meat during drying processing.

• Korean Journal of Food Science and Technology
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• v.17 no.4
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• pp.289-294
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• 1985

Total lipid (TL) content of sea squirt (Ureungsweng-ee), Halocynthia roretzi, and Mideuduck, Styela clava, were 2.0%, 2.1%, respectively. Reviewing the composition of each lipid fraction in total lipids of sea squirt and Mideuduck, it was found that contents of neutral lipids (NL) (36.6%, 36.3%) and phospholipids (PL) (46.2%, 44.5%) were high, while that of glycolipids (GL) (17.2%, 19.2%) was low. The NL of sea squirt and Mideuduck were mainly consisted of triglyceride (49.0%, 59.6%) and free sterol (25.8%, 22.0%), and followed by diglyceride (9.4%, 7.7%), monoglyceride (6.0%, 4.2%), free fatty acid (4.6%, 1.9%) and esterified sterol and hydrocarbon (5.2%, 4.4%). And main lipids in PL were phosphatidylcholine (48.6%, 46.7%) and phosphatidylethanolamine (32.4%, 35.0%), and followed by phosphatidylinositol (9.8%, 7.0%), phosphatidylserine (5.7%, 5.8%) and an unknown substance (3.5%, 5.5%). Fatty acid composition was not significantly different among TL, NL, PL and GL contained in sea squirt and Mideuduck. The major fatty acids of TL in sea squirt and Mideuduck were eicosapentaenoic (21.3%, 18.3%), docosahexaenoic (16.3%, 14.2%), palmitic (13.8, 16.3%) and oleic acid (8.5%, 7.0%), respectively. Fatty acid composition of PL and NL were similar to those of TL. In case of GL fraction the major fatty acids were gadoleic (15.7%, 14.7%), palmitic (13.5%, 14.7%), stearic (11.6%. 9.8%) and oleic acid (8.0%, 8.1%).

• Korean Journal of Veterinary Research
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• v.32 no.4
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• pp.531-542
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• 1992

The present study was performed to identify the factors influencing on early pregnacy and embryo implantation in rabbit. Serum, uterine fluid, and uterine tissue were collected on day 0, 3, 5, 7 and 9 of pregnancy. The intrauterine environment of receptive phase and refractory phase was compared by measuring protein synthetic capacity of endometrium, amino acid composition and concentrations of lipids(phospholipid, cholesterol). The results obstained were as follows : 1. The concentrations of total protein were significantly increased (p<0.01) on day 5 ($7.00{\pm}0.55$), 7($6.29{\pm}0.65$), and 9($6.34{\pm}0.61$), compared to those on day 0($5.50{\pm}0.12g/100m{\ell}$) in serum. The concentration of albumin on day 0 was $0.81{\pm}0.05$ and reached maximum on day 5 ($1.59{\pm}0.07g/100m{\ell}$) in serum. The concentrations of total protein were significantly increased(p<0.01) on day 5($1.56{\pm}0.10$), 7($1.99{\pm}0.22$), compared to those on day 0($0.38{\pm}0.02g/100m{\ell}$) in uterine fluid. The concentration of albumin on day 5($0.78{\pm}0.05g/100m{\ell}$) was higher than those on the other days in uterine fluid. 2. The incorporation rates of [$^3H$]-leucine into protein were significantly increased(p<0.01) on day 5 ($919.6{\pm}97.5$), 7($1445.4{\pm}95.9$) and 9($450.38{\pm}28.71$), compared to those on day 0($328.2{\pm}38.9cpm/mg$ protein) in endometrium. The incorporation rates in colehicine-treated endometrium on day 5 ($1341.9{\pm}73.8$), 7($1729.4{\pm}63.3cpm/mg$ protein) were significantly higher(p<0.01) than those on the other days. 3. The compositions of amino acid were not distinctly changed during early pregnancy in serum. The composition ratios of methione, lysine were distinctly decreased on day 3, compared to those on day 0 in uterine fluid. Those of glycine, alanine were increased on day 9, compared to those on other days but his tidine decreased in uterine fluid. 4. The concentrations of total phospholipid and total cholesterol were significantly decreased(p<0.01) on day 3($77.9{\pm}15.5$, $61.5{\pm}21.2$), compared to those on day 0($164.0{\pm}33.9$, $167.2{\pm}46.2mg/100m{\ell}$)in serum. The concentrations of total phospholipid and total cholesterol on day 9 ($47.3{\pm}13.4$, $37.7{\pm}9.6mg/100m{\ell}$) were significantly higher(p<0.01) than those on the other days in uterine fluid. 5. Total phopholipid/total cholesterol ratios were not significantly changed during early pregnancy in serum. However, total phospholipid/total cholesterol ratios on day 5 ($2.00{\pm}0.42$), 7 ($1.11{\pm}0.77$) and 9 ($1.47{\pm}0.30$) were higher than those on day 3($0.84{\pm}0.41$) in uterine fluid. 6. The concentrations of phosphatidylcholine and phosphatidylserine were significantly increased (p<0.01) on the other days, compared to those on day 0 during early pregnancy in serum. The concentrations of phosphatidylcholine were significantly increased(p<0.01),compared to those on day 0 and those of phosphatidyl-ethanolamine were consistently increased but not significant in early pregnancy in uterine fluid.