Squamous cell carcinoma (SCC) in head and neck show a variability in the response to chemotherapy, even when it present with similar histological tumor type, grade, and clinical stage. The purpose of present study it to identify predictive bio-marker for the sensitivity or resistance to conventional chemotherapeutic agents, 5-fluorouracil (5-FU) and Cisplatin Oral cancer cell lines were used in present study. MTT assay was performed to evaluate the sensitivity and/or resistance to 5-FU and Cisplatin. And RT-PCR was carried out for evaluation of the mRNA expressions of various genes associated with mutation, inflammation (COX pathway), cell cycle, senescence and extracellular matrix (ECM). The molecules which are correlated with the sensitivity to 5-FU are XPA, XPC, OGG, APEX, COX-2, PPAR, Cyclin E, Cyclin B1, CDC2, hTERT, hTR, TIMP-3, TIMP-4 and HSP47. And the molecules are correlated with the sensitivity to Cisplatin are COX-1, iNOS, eNOS, PCNA, collagen 1 and MMP-9. Taken together, when choosing the appropriate chemotherpeutic agents for patients, considering the molecules which are correlated or reversely correlated is helpful to choose the resonable agents for cancer patients.
An, Su-Yeon;Lee, Hoon-Taek;Kim, Suel-Kee;Yoon, Yong-Dal;Lee, Ho-Joon
Proceedings of the KSAR Conference
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2003.06a
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pp.31-31
/
2003
The incidence of reproductive abnormalities in the male has been reported to have increased during the past 50 years. These changes may be attributable to the presence of chemical with oestrogenic activity in our environment. Present study was carried out to determine the effects of maternal exposure to xenoestrogens on the testicular development and on the transcriptional expression of the steroidogenic enzyme and subunits of inhibin/activin in testis of male offspring. Pregnant female mice were administrated with 4-tert-octylphenol (OP; 2, 20, 200mg/kg), Bisphenol A (BPA; 2, 20, 200$\mu\textrm{g}$/kg), $\beta$-estradiol 17-valerate (EV; 2$\mu\textrm{g}$/kg) or vehicle (CV; corn oil) during gestational days 11 to 17. Offsprings were sacrificed on gestational day 18 (fetal 18) and neonatal day 7. Body weights were significantly increased in groups treated with all doses of OP and BPA. Maximum seminiferous tubules diameter on gestational day 18 were not changed in any treatment group, however, they were significantly increased on the neonatal day 7 in the group treated with low-dose of OP (2 mg/kg) and BPA (2 $\mu\textrm{g}$/kg). Increased expression of the P450$_{17a}$-hydroxylase dehydrogenase (P450$_{17a}$), 3$\beta$-hydroxylase dehydrogenase (3$\beta$-HSD), and 17$\beta$-hydroxylase dehydrogenase (17$\beta$-HSD) on gestational day 18 were observed in the groups treated with 2 or 20 mg/kg of OP. However, expression of the steroidogenic enzymes were not changed in the groups treated with all the doses of BPA. In contrast with the results from fetal testis, no expressional changes of these enzymes was found in all the OP-treated group and increased expression of inhibin/activin $\beta$B subunit mRNA were obseued in the 200 $\mu\textrm{g}$/kg BPA-treated group in the neonatal day 7. These results suggest that gestational exposure to low level of xenoestrogen causes a stimulatory effects on the transcriptional expressions of steroidogenic enzymes and subunits of inhibin/activin and on the seminiferous tubule development by their estrogen-like actions.ons.
This study was performed to investigate the effect of desalinated underground seawater (named as 'magma seawater', MSW) of Jeju Island in Korea on lipid metabolism and antioxidant activity. MSW was collected from underground of Han-Dong in Jeju Island, and freely given to high fat diet (HFD)-fed C57BL/6 mice for 10 weeks. Although there were no significant differences in the body weight changes and plasma lipid levels, hepatic triglyceride levels were significantly lower in the MSW group than in the normal tap water (TW)-drunken control group. Furthermore, the activity of fatty acid synthase (FAS) was significantly decreased and carnitine palmitoyltransferase (CPT) activity was increased in MSW group compared to TW group. Similarly, real-time PCR analysis revealed that mRNA expressions of lipogenic genes were lowered in MSW groups compared to the control group. In a morphometric observation on the liver tissue, accumulation of fats was remarkably reduced in MSW group. Meanwhile, in vitro assay, tree radical scavenging activity measured by using diphenylpicrylhydrazyl (DPPH) was increased in MSW group. The 2'-7'-dichlorofluorescein diacetate (DCF-DA) staining followed with fluorescent microscopy showed a low intensity of fluorescence in MSW-treated HepG2 cells, compared to TW-treated HepG2 cells, which indicated that the production of reactive oxygen species by tert-butyl hydroperoxide (t-BHP) in HepG2 cells was decreased by MSW treatment. The antioxidant effect of MSW on t-BHP-induced oxidative stress in HepG2 cells was supported by the increased activities of intracellular antioxidant enzymes such as catalase and glutathione reductase. From these results, we speculate that MSW has an inhibitory effect on lipogenesis in liver and might play a protective role against cell damage by t-BHP-induced oxidative stress.
Choi, Shinkyu;Na, Hye-Young;Kim, Ji Aee;Cho, Sung-Eun;Suh, Suk Hyo
The Korean Journal of Physiology and Pharmacology
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v.17
no.3
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pp.181-187
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2013
Reactive oxygen species (ROS) are generated in various cells, including vascular smooth muscle and endothelial cells, and regulate ion channel functions. $K_{Ca}3.1$ plays an important role in endothelial functions. However, the effects of superoxide and hydrogen peroxide radicals on the expression of this ion channel in the endothelium remain unclear. In this study, we examined the effects of ROS donors on $K_{Ca}3.1$ expression and the $K^+$ current in primary cultured human umbilical vein endothelial cells (HUVECs). The hydrogen peroxide donor, tert-butyl hydroperoxide (TBHP), upregulated $K_{Ca}3.1$ expression, while the superoxide donors, xanthine/xanthine oxidase mixture (X/XO) and lysophosphatidylcholine (LPC), downregulated its expression, in a concentration-dependent manner. These ROS donor effects were prevented by antioxidants or superoxide dismustase. Phosphorylated extracellular signal-regulated kinase (pERK) was upregulated by TBHP and downregulated by X/XO. In addition, repressor element-1-silencing transcription factor (REST) was downregulated by TBHP, and upregulated by X/XO. Furthermore, $K_{Ca}3.1$ current, which was activated by clamping cells with 1 ${\mu}M$$Ca^{2+}$ and applying the $K_{Ca}3.1$ activator 1-ethyl-2-benzimidazolinone, was further augmented by TBHP, and inhibited by X/XO. These effects were prevented by antioxidants. The results suggest that hydrogen peroxide increases $K_{Ca}3.1$ expression by upregulating pERK and downregulating REST, and augments the $K^+$ current. On the other hand, superoxide reduces $K_{Ca}3.1$ expression by downregulating pERK and upregulating REST, and inhibits the $K^+$ current. ROS thereby play a key role in both physiological and pathological processes in endothelial cells by regulating $K_{Ca}3.1$ and endothelial function.
Background: Oxidative stress-induced cardiomyocytes apoptosis is a key pathological process in ischemic heart disease. Glutathione reductase (GR) reduces glutathione disulfide to glutathione (GSH) to alleviate oxidative stress. Ginsenoside Rb1 (GRb1) prevents the apoptosis of cardiomyocytes; however, the role of GR in this process is unclear. Therefore, the effects of GRb1 on GR were investigated in this study. Methods: The antiapoptotic effects of GRb1 were evaluated in H9C2 cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, annexin V/propidium iodide staining, and Western blotting. The antioxidative effects were measured by a reactive oxygen species assay, and GSH levels and GR activity were examined in the presence and absence of the GR inhibitor 1,3-bis-(2-chloroethyl)-1-nitrosourea. Molecular docking and molecular dynamics simulations were used to investigate the binding of GRb1 to GR. The direct influence of GRb1 on GR was confirmed by recombinant human GR protein. Results: GRb1 pretreatment caused dose-dependent inhibition of tert-butyl hydroperoxide-induced cell apoptosis, at a level comparable to that of the positive control N-acetyl-L-cysteine. The binding energy between GRb1 and GR was positive (-6.426 kcal/mol), and the binding was stable. GRb1 significantl reduced reactive oxygen species production and increased GSH level and GR activity without altering GR protein expression in H9C2 cells. Moreover, GRb1 enhanced the recombinant human GR protein activity in vitro, with a half-maximal effective concentration of ≈2.317 μM. Conversely, 1,3-bis-(2-chloroethyl)-1-nitrosourea co-treatment significantly abolished the GRb1's apoptotic and antioxidative effects of GRb1 in H9C2 cells. Conclusion: GRb1 is a potential natural GR agonist that protects against oxidative stress-induced apoptosis of H9C2 cells.
Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of $17{\beta}$-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with $0.001-100{\mu}M$ of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P < 0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P < 0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P < 0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P < 0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P < 0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P < 0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P < 0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.
Kim, Kwang Yeon;Lee, Seung Jin;Jee, Seon Young;Bae, Su Jin;Song, Yu Rim;Yun, Un-Jung;Bak, Seonbeen;Song, Jong Kuk;Son, Tae Jin;Son, Jae-Dong;Kim, Woo Hyun;Yang, Ju Hye;Park, Sun Dong;Kim, Sang Chan;Kim, Young Woo;Park, Kwang-Il
Herbal Formula Science
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v.29
no.2
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pp.71-80
/
2021
Objectives : This study investigated cellular-protective effects of Nardotidis seu Sulculii Concha water extract (NSCE) against oxidative stress induced by arachidonic acid (AA)+iron or tert-butylhydroperoxide (tBHP). Methods : In vitro, MTT assay was assessed for cell viability, and immunoblotting analysis was performed to detect expression of AMP-activated kinase (AMPK) signaling pathway and autophagy related proteins. In vivo, mice were orally administrated with the aqueous extract of NSCE of 500 mg/kg for 3 days, and then injected with CCl4 0.5 mg/kg body weight to induce acute damage. The level of liver damage was measured by serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) analysis. Results : Treatment with NSCE inhibited cell death induced by AA+iron and tBHP. NSCE induced the phosphorylation of AMPK, and this compound also induced the phosphorylation of LKB1, an upstream kinase of AMPK, and Acetyl-CoA carboxylase (ACC), a primary downstream target of AMPK. NSCE increased the protein levels of autophagic markers (LC3II and beclin-1) and decreased the phosphorylation of mammalian target of rapamycin (mTOR) and simultaneously increased the phosphorylation of unc-51-like kinase-1 (ULK-1) in time-dependent manner. Conclusions : NSCE has the ability 1) to protect cells against oxidative stress induced by AA+iron or tBHP. NSCE 2) to activate AMP-activated protein kinase (AMPK), and 3) to regulate autophagy, an important regulator in cell survival.
Journal of The Korean Society of Integrative Medicine
/
v.12
no.1
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pp.1-10
/
2024
Purpose: Lycopene is abundantly contained in Tomatoes and is known for diverse biological activities such as antioxidant, anti-inflammatory, and anticancer effects. In this study, the antioxidative potential of lycopene was investigated through the induction of hemeoxygenase (HO)-1 by nuclear factor-erythroid 2 p45-related factor2 (Nrf2) and upstream signaling molecules, mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/Aktin RAW 264.7 cells. Methods: The antioxidative potential of lycopene against oxidative stress and its molecular mechanisms were determined by the cell viability assay, intracellular reactive oxygen species (ROS) formation assay, and Western blot analysis in RAW 264.7 cells. Results: Lycopene treatment significantly attenuated tert-butyl hydroperoxide (t-BHP) induced intracellular ROS formation in a dose-dependent manner without any cytotoxicity. In addition, 50 µM of lycopene for 6 h treatment induced potent HO-1 expression and its transcription factor, Nrf2. MAPK and PI3K/Aktwere also analyzed due to their critical roles in the regulation of cellular redox homeostasis against oxidative damage. As a result, phosphorylation of extracellular regulated kinase (ERK) was significantly induced by lycopene treatment while the activated status of c-Jun NH2-terminal kinase (JNK), p38, and Akt, were not given any effect. To confirm the antioxidative mechanism of HO-1 mediated by ERK activation, each selective inhibitor was employed in a protection assay, in which oxidative damage occurred by t-BHP. Lycopene, SnPP, and CoPP treatments reflected accelerated HO-1 expression could be a protective role against oxidative damage-initiated cell death. A selective inhibitor for ERK significantly inhibited the lycopene-induced cytoprotective effect but selective inhibitors for other signaling molecules did not attenuate the rate of t-BHP-induced cell death. Conclusion: In conclusion, lycopene potently scavenged intracellular ROS formation and enhanced the HO-1 mediated antioxidative potential through the modulation of Nrf2, MAPK signaling pathway in RAW 264.7 cells.
Laboratory and greenhouse studies were conducted to determine differential sensitivities on absorption of $^{14}$ C-oxyfluorfen and the anatomical responses in wheat and barley to protoporphyrinogen oxidase-inhibiting herbicides [oxyfluorfen (2-chloro-1- (3-ethoxy -nitrophen-oxy)-4-(trifluoromethyl) benzene, acifluorfen(5-[2-chloro-4-(trifluoromethyl) phenoxy]-2-nitro-benzoic acid), bifenox(methyl-5-(2, 4-dichlorophenoxy)-2-nitrobenzoate) and oxadiazon(5-tert-butyl-3-(2, 4-dichloro-5-isopropoxyphenyl)-1, 3, 4-oxadiazol-2-one)]. I$_{50}$ value of the tolerant wheat cultivars to oxyfluorfen was about 10$^{-4}$ , whereas that of the susceptible barley cultivars was about 10$^{-6}$ M, showing significant difference between the two groups. When foliage were applied with acifluorfen, bifenox or oxadiazon, the oxyfluorfen-tolerant wheat showed less decreased in shoot fresh weight and chlorophyll content than the susceptible barley. Also, when soil-applied with these herbicides test plants showed similar tendency in foliar application. Electrolyte leakage from the tissue treated with these compounds was the more influenced in the barley than the wheat. Malondialdehyde(MDA) production as index of lipid peroxidation was greater in the barley than the wheat by treatment of these compounds. Therefore, the differential sensitivities of wheat and barley to protoporphyrinogen oxidaseinhibiting herbicides was showed by our greenhouse and in vitro experiment. The absorption rates of $^{14}$ C-oxyfluorfen were higher in the barley than the wheat. And this tendency was showed appararitly difference by increase of treatment durations. After the oxfluorfen and oxadiazon treatment, the tolerant wheat did not show the structural damage in leaf surface, but the susceptible barley was damaged in the leaf waxy layer. However, the acifluorfen and bifenox treatment showed no difference between wheat and barley. The anatomical changes by these compounds treatment were not observed in the tolerant wheat but epidermal cell and mesophyll cell were highly broken in the susceptible barley.
This experiment was conducted to investigate the protoporphyrin Ⅸ (PPIX)accumulation, activity of antioxidative enzymes and contents of antioxidant in tolerant-wheat and susceptible-barley to protoporphyrinogen oxidase (Protox) inhibiting-herbicides [oxyfluorfen(2-chloro-l-(3-ethoxy-nitrophenoxy-4-(trifluoromethyl) benzene, acifluorfen (5-[2-chloro-4-(trifl-uoromethyl) phenoxy]-2-nitrobenzoic acid), bifenox(methyl-5-(2, 4-dichlorophenoxy) 2-nitroben-zoate), and oxadiazon (5-tert-butyl-3-(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one)]. The tolerant-wheat and susceptible-barley were soaked in these compounds at 10$^{-6}$ M for 2hrs and exposed to light for 2,4,6 or 8hrs to investigate change of the activity of antioxidative enzymes. The activities of monodehydroascorbate reductase(MDAR), catalase(CAL) and superoxide dismutase(SOD) were lower in the barley than in the wheat after the treatement of these compounds. The activity of peroxidase(POX) was lower in the barley than in the wheat at 8hrs after the treatment of oxyfluorfen but other compounds showed no difference in activity in wheat and barley. The activity of glutathione reductase(GR) was increased in wheat and barley according as hours of treatment of these compounds became increased but its activity was no difference between wheat and barley. In the case of the content of vitamin C due to the treatment of these compounds, the wheat decreased less than the barley. After the treatment of oxyfluorfen the content of vitamin E in the wheat was higher than in the barley but other compounds didn't have any difference between wheat and barley. And after the treatment of acifluorfen the content of carotenoid was greater in the wheat than in the barley but other compounds didn't have any difference between wheat and barley. The content of glutathione (GSH, GSSG) was greater in the barley than in the wheat. The content of protoporphyrin Ⅸ (PPIX) accumulation by the treatments of these compounds was more in the barley than in the wheat. Especially, the treatment of oxyfluorfen and acifluorfen were more accumulated 2.3 and 1.3 fold in the barley than in the wheat, respectively.
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