• Environmental Mutagens and Carcinogens
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• v.22 no.4
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• pp.259-265
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• 2002

Phthalate analogues are a plasticizer and solvent used in industry. Phthalates were reported to be a potential carcinogen classified in the category of suspected endocrine disruptors. Most common human exposure to these compounds may occur with contaminated food. They may migrate into food from plastic wrap or may enter food from general environmental contamination. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of phthalates that possibly threaten the public health. Concern about their use has been mounting. To screen and elucidate the endocrine disrupting activity and their mechanism of phthalate analogues, first of all, E-screen assay was performed in MCF7 human breast cancer cells with seven phthalate analogues. In this cell proliferation assay, only dibutyl phthalate (DBP) showed weak estrogenic activity. Also the yeast-based transcription assay to assess the interactions of DBP with the estrogen, androgen, and progesterone receptors was conducted. DBP in the concentration ranges from 10$^{-16}$ to 10$^{-11}$ M was active in the estrogen transcriptional assay, but it did not show the effect on $\beta$-galactosidase activity in the progesterone and androgen transcriptional assays. These data indicate that DBP shows estrogenic potential and can be classified as weak and/or suspected endocrine disrupting chemicals.

• Journal of Microbiology and Biotechnology
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• v.18 no.8
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• pp.1475-1481
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• 2008

B3 antibody specifically binds the $Lewis^Y$-related carbohydrate antigen of many carcinomas, and it is used as a model antibody in this study. In a previous study, the Fab fragment of the antibody was fused to a 38 kDa truncated form of Pseudomonas exotoxin A, PE38, to make Fab-PE38, where PE38 is fused to the Fd fragment of the Fab domain. This parent monomer molecule, Fab-PE38, had no cysteine in the hinge region, and it could not make a disulfide bond to form a disulfide bond bridged homodimer. In this study, we constructed three different kinds of divalent Fab-toxin fusion homodimers where the toxin is fused to the light chain of Fab, $(Fab-PE38fl)_2$. In addition to the PE38 toxin fused to the light chain, these three molecules have different hinge sequences hi, h2, and h3 making Fabh1-, Fabh2-, and Fabh3-PE38fl monomers, respectively. These hinges contain only one cysteine on different positions of the hinge sequence. The disulfide bond between the hinge region of two monomers forms homodimers $(Fabh1-PE38fl)_2$, $(Fabh2-PE38fl)_2$, and $(Fabh3-PE38fl)_2$. The refolding yields of these dimers were 5-16-fold higher than a previously constructed dimer where the PE38 was fused to the Fd fragment $(Fabh2-PE38)_2$ [8]. Our data suggest that the steric repulsion between the two PE38s in $(Fabh1-PE38)_2$ during disulfide bridge formation is relieved by fusing it at the end of the light chain. The best cytotoxicity value of these dimers showed about 2.5-fold higher on an MCF7 cell line than that of the monovalent reference molecule in ng/ml scale, which is 15-fold higher in pM scale.

• CELLMED
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• v.3 no.4
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• pp.32.1-32.6
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• 2013

Plectranthus amboinicus Benth (Lamiaceae) is a medicinal plant native to India, and its leaves are widely used in several traditional medicinal preparations. The purpose of this study was to detect and quantify phenolics present in ethyl acetate and acetone extracts of P. amboinicus leaves, and evaluate their antioxidant, antibacterial, antimutagenic and anticancer activities. The HPLC chromatograms of crude leaf extracts indicated the presence of phenolics like caffeic acid, coumaric acid, rutin, quercetin and gallic acid, which were present in the range of 0.01 - 1.41 mg/g in ethyl acetate and 0.03 - 1.93 mg/g in the acetone extract. The acetone extract showed statistically (p < 0.05) higher antioxidant activity ($IC_{50}$, 99.59 ${\mu}g/ml$) than ethyl acetate extract ($IC_{50}$, 149.96 ${\mu}g/ml$). Statistically (p < 0.05) higher antimutagenicity was shown by acetone extract (46.16%) as compare to ethyl acetate extract (12.16%) at 500 ${\mu}g/plate$ concentration. The acetone extract showed higher antibacterial activity than ethyl acetate extract, and both the extracts showed highest activity against B. cereus (375 and 625 ${\mu}g/ml$, respectively) and lowest activity against Y. enterocolitica (1000 and 1125 ${\mu}g/ml$, respectively). Both the extracts also showed inhibitory effect on cancer cell lines HCT-15 and MCF-7. These results suggest that the leaves of P. amboinicus possess various biological activities, and validate the traditional use of the leaves of P. amboinicus against cold, infection and ulceration.

• Environmental Analysis Health and Toxicology
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• v.19 no.2
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• pp.207-216
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• 2004

유기염소계 농약은 화학적으로 안정하고, 지용성이 크며 체내 대사에 대해 저항성을 가지고 있어서 인체 및 생태계의 생물체 중 축적성이 매우 크다. 또한 대부분 내분비계 장애물질로 분류되어 있어 관심의 대상이 되고 있다. 유기염소계 농약은 잔류성으로 인하여 사용이 금지되었음에도 불구하고 인체에서 검출되고 있으며 한국인의 지방조직을 분석한 결과 9종의 유기염소계 농약이 주로 축적되어 있음이 보고된 바 있다. 본 연구에서는 이들 유기염소계 농약의 에스트로겐 활성을 MCF-7 BUS cell을 이용한 E-screen assay competitive binding assay 및 pS$_2$ gene experession assay에 의해 조사하였다. o,p'-DDT,p, p'-DDT,p,p'-DDD,p,p'-DDE등 4종의 유기염소계 농약은 에스트로겐 수용체에 대한 ligand의존적인 작용기전에 의해 에스트로겐 활성을 가지며, $\alpha$-, $\beta$-, ${\gamma}$-, $\delta$-BHC, dieldrin등 5종의 유기염소계 농약은 lingand-비의존적 작용기전에 의해 에스트로겐 활성을 보였다. 또한 이들 유기염소계 농약을 혼합 투여하여 에스트로겐 활성을 관찰한 결과 DDT류의 경우에는 단독투여시 보다 그 대사체와 혼합 투여할 때 에스트로겐 활성에 상승적 효과가 나타났으며 o,p'-와 p,p'-DDT의 두 이성질체를 혼합 투여할 경우가 단독 투여시 보다 상승적 효과가 나타났다. 따라서 지방조직에서 검출되는 유기염소계농약은 상호작용에 의해 개별 물질이 나타내는 내분비계장애작용 보다 실제로는 강한 효과를 나타낼 것으로 추정되었다.

• Environmental Engineering Research
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• v.21 no.2
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• pp.188-195
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• 2016

$17{\alpha}$-Ethynylestradiol (EE2) has gotten growing concerns due to its widely detected in the environment and high estrogenic potency. However, the knowledge on the photochemical behaviors of EE2 in natural waters is still limited. Herein, the photodegradation and estrogenic potency variation of EE2 induced by nitrate were studied using a sunlight simulator consisted by a 300 W medium pressure mercury lamp and 290 nm cut-off filters. It was found that EE2 could be photodegraded at a rate of $0.0193h^{-1}$ in pure aqueous solutions, and the photodegradation of EE2 could be significantly promoted by nitrate. The photodegradation removal rate of EE2 was increased from 9% in Milli-Q water to 85% in 2.0 mM nitrate solutions. Reactive species scavenging experiments demonstrated that the photogenerated $HO{\bullet}$ contributed about 55% to EE2 degradation. Fe(III), Cl- and dissolved humic acid (DHA) could inhibit the photodegradation of EE2 by competing the incident light and photogenerated $HO{\bullet}$, while $HCO_3{^-}$ had no influence on EE2 photodegradation. EE2 was determined to be phototransformed into organic chemicals without estrogenic potency by GC-MS and MCF-7 cell proliferation toxicity tests. These findings could extend our knowledge on the photochemical behaviors of steroid estrogens and provide information for ecological risk assessment.

• Korean Journal of Pharmacognosy
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• v.45 no.2
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• pp.174-180
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• 2014

D-Pinitol, an anti-diabetic substance, is a naturally occurring compound found in legumes. In this study, we investigated the inhibitory effect of D-pinitol on growth and recurrence of breast cancer. When D-pinitol was treated on MDA-MB-231 or MCF-7 breast cancer cells, it was observed that the viability of the two cancer cell lines was reduced in MTT assay. In order to examine the effect on the growth of breast tumor, mouse xenograft assay was carried out. On day 0, nine millions cells of MDA-MB-231 were injected subcutaneously into nude mouse and D-pinitol was administered orally at the dose of 500 mg/kg or 1000 mg/kg body weight for consecutive 45 days. Tumor size was reduced in dose-dependent manner upto 95.4% in 1000 mpk-treated group, compared with the non-treated control group. When D-pinitol was co-administrated with $4{\mu}g$ of doxorubicin, recurrence of breast tumor was delayed by two weeks, compared with the mouse group of doxorubicin monotherapy. Consistent with this data, it was observed that the population of cancer stem cells (CSCs), responsible for recurrence of cancer, within tumor mass was significantly reduced. Taken together, D-pinitol inhibits the growth of breast cancer and relapse of the tumor by suppressing the proliferation of CSCs.

• Genomics & Informatics
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• v.17 no.3
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• pp.26.1-26.12
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• 2019

Identification of fusion gene is of prominent importance in cancer research field because of their potential as carcinogenic drivers. RNA sequencing (RNA-Seq) data have been the most useful source for identification of fusion transcripts. Although a number of algorithms have been developed thus far, most programs produce too many false-positives, thus making experimental confirmation almost impossible. We still lack a reliable program that achieves high precision with reasonable recall rate. Here, we present FusionScan, a highly optimized tool for predicting fusion transcripts from RNA-Seq data. We specifically search for split reads composed of intact exons at the fusion boundaries. Using 269 known fusion cases as the reference, we have implemented various mapping and filtering strategies to remove false-positives without discarding genuine fusions. In the performance test using three cell line datasets with validated fusion cases (NCI-H660, K562, and MCF-7), FusionScan outperformed other existing programs by a considerable margin, achieving the precision and recall rates of 60% and 79%, respectively. Simulation test also demonstrated that FusionScan recovered most of true positives without producing an overwhelming number of false-positives regardless of sequencing depth and read length. The computation time was comparable to other leading tools. We also provide several curative means to help users investigate the details of fusion candidates easily. We believe that FusionScan would be a reliable, efficient and convenient program for detecting fusion transcripts that meet the requirements in the clinical and experimental community. FusionScan is freely available at http://fusionscan.ewha.ac.kr/.

• Biomolecules & Therapeutics
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• v.20 no.3
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• pp.326-331
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• 2012

We recently reported that Val-SN-38, a novel valine ester prodrug of SN-38, had greatly improved the intracellular accumulation of SN-38 in MCF-7 cell line, probably through enhanced uptake via amino acid transporters. In the present study, the efficacy of Val-SN-38 was further investigated both in vitro and in vivo. It was found that the in vitro cytotoxic effect of Val-SN-38 was similar to that of SN-38. Moreover, Val-SN-38 exhibited an equal potency to that of SN-38 in survival experiments in vivo. Because these results seemed to be contrary to the previous finding, further investigation was performed to find out the underlying cause of the contradiction. As only the lactone form is known to have cytotoxic activity, the proportion of lactone in Val-SN-38 and SN-38 was determined, but no differences were found. However, it turned out that Val-SN-38 had poor stability compared with SN-38, which resulted in a decrease in beneficial efficacy for Val-SN-38. Overall, the present study showed that a valine-added prodrug approach could be advantageous provided that the stability of the compound can be ensured. We believe this is a noteworthy study that unravels the discrepancy between intracellular accumulation and efficacy of valine-added prodrug.

• Molecules and Cells
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• v.45 no.6
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• pp.425-434
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• 2022

The post-translational modification (e.g., phosphorylation) of estrogen receptor α (ERα) plays a role in controlling the expression and subcellular localization of ERα as well as its sensitivity to hormone response. Here, we show that ERα is also modified by UFM1 and this modification (ufmylation) plays a crucial role in promoting the stability and transactivity of ERα, which in turn promotes breast cancer development. The elevation of ufmylation via the knockdown of UFSP2 (the UFM1-deconjugating enzyme in humans) dramatically increases ERα stability by inhibiting ubiquitination. In contrast, ERα stability is decreased by the prevention of ufmylation via the silencing of UBA5 (the UFM1-activating E1 enzyme). Lys171 and Lys180 of ERα were identified as the major UFM1 acceptor sites, and the replacement of both Lys residues by Arg (2KR mutation) markedly reduced ERα stability. Moreover, the 2KR mutation abrogated the 17β-estradiol-induced transactivity of ERα and the expression of its downstream target genes, including pS2, cyclin D1, and c-Myc; this indicates that ERα ufmylation is required for its transactivation function. In addition, the 2KR mutation prevented anchorage-independent colony formation by MCF7 cells. Most notably, the expression of UFM1 and its conjugating machinery (i.e., UBA5, UFC1, UFL1, and UFBP1) were dramatically upregulated in ERα-positive breast cancer cell lines and tissues. Collectively, these findings implicate a critical role attributed to ERα ufmylation in breast cancer development by ameliorating its stability and transactivity.

• Journal of Microbiology and Biotechnology
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• v.32 no.9
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• pp.1195-1208
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• 2022

Silver nanoparticles (AgNPs) have potential applications in medicine, photocatalysis, agriculture, and cosmetic fields due to their unique physicochemical properties and strong antimicrobial activity. Here, AgNPs were synthesized using actinobacterial SL19 strain, isolated from acidic forest soil in Poland, and confirmed by UV-vis and FTIR spectroscopy, TEM, and zeta potential analysis. The AgNPs were polydispersed, stable, spherical, and small, with an average size of 23 nm. The FTIR study revealed the presence of bonds characteristic of proteins that cover nanoparticles. These proteins were then studied by using liquid chromatography with tandem mass spectrometry (LC-MS/MS) and identified with the highest similarity to hypothetical protein and porin with molecular masses equal to 41 and 38 kDa, respectively. Our AgNPs exhibited remarkable antibacterial activity against Escherichia coli and Pseudomonas aeruginosa. The combined, synergistic action of these synthesized AgNPs with commercial antibiotics (ampicillin, kanamycin, streptomycin, and tetracycline) enabled dose reductions in both components and increased their antimicrobial efficacy, especially in the case of streptomycin and tetracycline. Furthermore, the in vitro activity of the AgNPs on human cancer cell lines (MCF-7, A375, A549, and HepG2) showed cancer-specific sensitivity, while the genotoxic activity was evaluated by Ames assay, which revealed a lack of mutagenicity on the part of nanoparticles in Salmonella Typhimurium TA98 strain. We also studied the impact of the AgNPs on the catalytic and photocatalytic degradation of methyl orange (MO). The decomposition of MO was observed by a decrease in intensity of absorbance within time. The results of our study proved the easy, fast, and efficient synthesis of AgNPs using acidophilic actinomycete SL19 strain and demonstrated the remarkable potential of these AgNPs as anticancer and antibacterial agents. However, the properties and activity of such particles can vary by biosynthesized batch.