Identification of Biomarkers for Breast Cancer Using Databases

Eunhye Lee,,Aree Moon 대한암예방학회 2016 Journal of cancer prevention Vol.21 No.4

Breast cancer is one of the major causes of cancer death in women. Many studies have sought to identify specific molecules involved in breast cancer and understand their characteristics. Many biomarkers which are easily measurable, dependable, and inexpensive, with a high sensitivity and specificity have been identified. The rapidly increasing technology development and availability of epigenetic informations play critical roles in cancer. The accumulated data have been collected, stored, and analyzed in various types of databases. It is important to acknowledge useful and available data and retrieve them from databases. Nowadays, many researches utilize the databases, including The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), Surveillance, Epidemiology and End Results (SEER), and Embase, to find useful informations on biomarkers for breast cancer. This review summarizes the current databases which have been utilized for identification of biomarkers for breast cancer. The information provided by this review would be beneficial to seeking appropriate strategies for diagnosis and treatment of breast cancer.

Tumor-associated macrophages secrete CCL2 and induce the invasive phenotype of human breast epithelial cells through upregulation of ERO1-α and MMP-9

Lee, Seungeun,Lee, Eunhye,Ko, EunYi,Ham, Mina,Lee, Hye Min,Kim, Eun-Sook,Koh, Minsoo,Lim, Hyun Kyung,Jung, Joohee,Park, So Yeon,Moon, Aree Elsevier 2018 Cancer letters Vol.437 No.-

<P><B>Abstract</B></P> <P>Tumor-associated macrophages (TAMs) are major components of tumor microenvironment that promote invasion and metastasis of cancer cells. In this study, we investigated the effect of TAMs on phenotypic conversion of non-neoplastic MCF10A human breast epithelial cells using an indirect co-culture system. Co-culture with TAMs induced epithelial-to-mesenchymal transition, invasive phenotype, and MMP-9 upregulation in MCF10A cells. Comparative proteomic analysis revealed that endoplasmic reticulum oxidoreductase (ERO)1-α was increased in MCF10A cells co-cultured with TAMs compared to that in mono-cultured cells. ERO1-α was crucial for TAMs-induced invasive phenotype and MMP-9 upregulation involving transcription factors c-fos and c-Jun. Cytokine array analysis showed that levels of interleukin (IL)-6, C-X-C motif ligand (CXCL)1, C-C motif ligand (CCL)2, growth-regulated protein (GRO), IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) were increased in conditioned media of co-cultured cells. Among these cytokines increased in conditioned media of co-cultured cells, CCL2 was secreted from TAMs, leading to induction of ERO1-α, MMP-9 upregulation, and invasiveness in MCF10A cells. Our findings elucidated a molecular mechanism underlying the aggressive phenotypic change of non-neoplastic breast cells by co-culture with TAMs, providing useful information for prevention or treatment of recurrent breast cancer.</P> <P><B>Highlights</B></P> <P> <UL> <LI> TAMs induced invasion in MCF10A cells non-neoplastic human breast epithelial cells. </LI> <LI> ERO1-α was increased in MCF10A cells co-cultured with TAM. </LI> <LI> ERO1-α was crucial for the TAMs-induced invasive phenotype and MMP-9 upregulation. </LI> <LI> CCL2 was secreted from TAMs co-cultured with MCF10A cells. </LI> <LI> CCL2 induced ERO1-α, resulting MMP-9 upregulation and invasiveness in MCF10A cells. </LI> </UL> </P>

CNBP acts as a key transcriptional regulator of sustained expression of interleukin-6

Lee, Eunhye,Lee, Taeyun A.,Kim, Ji Hyun,Park, Areum,Ra, Eun A.,Kang, Sujin,Choi, Hyun jin,Choi, Junhee L.,Huh, Hyunbin D.,Lee, Ji Eun,Lee, Sungwook,Park, Boyoun Oxford University Press 2017 Nucleic acids research Vol.45 No.6

<P><B>Abstract</B></P><P>The transcription of inflammatory genes is an essential step in host defense activation. Here, we show that cellular nucleic acid-binding protein (CNBP) acts as a transcription regulator that is required for activating the innate immune response. We identified specific CNBP-binding motifs present in the promoter region of sustained inflammatory cytokines, thus, directly inducing the expression of target genes. In particular, lipopolysaccharide (LPS) induced <I>cnbp</I> expression through an NF-κB-dependent manner and a positive autoregulatory mechanism, which enables prolonged <I>il-6</I> gene expression. This event depends strictly on LPS-induced CNBP nuclear translocation through phosphorylation-mediated dimerization. Consequently, <I>cnbp</I>-depleted zebrafish are highly susceptible to <I>Shigella flexneri</I> infection <I>in vivo</I>. Collectively, these observations identify CNBP as a key transcriptional regulator required for activating and maintaining the immune response.</P>

<i>In vivo</i> antitumor effects of chitosan-conjugated docetaxel after oral administration

Lee, Eunhye,Kim, Hyungjun,Lee, In-Hyun,Jon, Sangyong Elsevier 2009 Journal of controlled release Vol.140 No.2

<P><B>Abstract</B></P><P>The purpose of this study is to evaluate <I>in vivo</I> antitumor efficacy and subacute toxicity of docetaxel (DTX) prodrug comprising a conjugate between DTX and low molecular weight chitosan (LMWC) after oral administration. DTX was covalently attached to LMWC via a cleavable linker so as to be released from LMWC-DTX conjugate in body. <I>In vitro</I> cytotoxicity of LMWC-DTX conjugate was evaluated by MTT assay against two human cancer cell lines, showing similar IC<SUB>50</SUB> values to the parent DTX. The pharmacokinetic data of the conjugate after oral administration revealed that half-life in blood circulation was increased by ~15-fold and AUC<SUB>(0–∞)</SUB> was 3.8–6.2 times higher in comparison with the intravenously injected DTX (i.v.). <I>In vivo</I> antitumor efficacy was evaluated in nude mice bearing human non-small cell lung carcinoma (NCI-H358) and glioblastoma (U87MG), respectively. The orally administered LMWC-DTX conjugate (10mg DTX equivalent/kg) showed comparable antitumor efficacy to the same dose of DTX (i.v.) for both NCI-H358 and U87MG models, but revealed much lower subacute toxicity as seen in body weight loss and hematological toxicity.</P> <P><B>Graphical abstract</B></P><P><ce:figure></ce:figure></P>

High Performance Liquid Chromatographic Method for Determination of Metazosulfuron Residue in Representative Crops

Lee, Hyeri,Kim, Eunhye,Lee, Young Deuk,Kim, Jeong-Han The Korean Society of Environmental Agriculture 2013 한국환경농학회지 Vol.32 No.2

BACKGROUND: This study was performed to develop a single residue analytical method for new herbicide metazosulfuron in crops. METHODS AND RESULTS: Brown rice, apple, mandarin, Kimchi cabbage and soybean were selected as representative crops, and clean-up system, partition solvent and extraction solvent were optimized. Instrumental limit of quantitation (ILOQ), linearity of calibration curve and method limit of quantitation (MLOQ) were determined based on the chromatography and whole procedures. For recovery tests, brown rice, apple, mandarin, Kimchi cabbage and soybean samples were macerated and fortified with metazosulfuron standard solution at three levels (MLOQ, 10 MLOQ and 100 MLOQ). And then those were extracted with acetonitrile, concentrated, and partitioned with ethyl acetate. Then the extracts were concentrated again and cleaned-up through $NH_2$ (aminopropyl) SPE cartridge with acetone : dichloromethane (1% acetic acid) (20 : 80, v/v) before concentration and analysis with HPLC. CONCLUSION(S): ILOQ of metazosulfuron was 2 ng (S/N${\geq}$10) and good linearity was achieved between 0.05 and 12.5 mg/Kg of metazosulfuron standard solutions, with coefficients of determination of 0.9999. MLOQ was 0.02 mg/Kg. Good recoveries from 74.1 to 116.9% with coefficients of variation (C.V.) of less than 10% were obtained, regardless of sample type, which satisfies the criteria of Korea Food and Drug Administration (KFDA). Those results were reconfirmed with LC-MS (SIM). The method established in this study is simple, economic and efficient to be applied to most of crops as an official and general method for residue analysis of metazosulfuron.

Pyrazole C-region analogues of 2-(3-fluoro-4-methylsulfonylaminophenyl)propanamides as potent TRPV1 antagonists

Lee, Sunho,Kim, Changhoon,Ann, Jihyae,Thorat, Shivaji A.,Kim, Eunhye,Park, Jongmi,Choi, Sun,Blumberg, Peter M.,Frank-Foltyn, Robert,Bahrenberg, Gregor,Stockhausen, Hannelore,Christoph, Thomas,Lee, Jee Pergamon Press 2017 Bioorganic & medicinal chemistry letters Vol.27 No.18

<P><B>Abstract</B></P> <P>A series of 1-substituted 3-(<I>t</I>-butyl/trifluoromethyl)pyrazole C-region analogues of 2-(3-fluoro-4-methylsulfonamidophenyl)propanamides were investigated for <I>h</I>TRPV1 antagonism. The structure activity relationship indicated that the 3-chlorophenyl group at the 1-position of pyrazole was the optimized hydrophobic group for antagonistic potency and the activity was stereospecific to the <I>S</I>-configuration, providing exceptionally potent antagonists <B>13<I>S</I> </B> and <B>16<I>S</I> </B> with <I>K<SUB>i(CAP)</SUB> </I> =0.1nM. Particularly significant, <B>13<I>S</I> </B> exhibited antagonism selective for capsaicin and NADA and not for low pH or elevated temperature. Both compounds also proved to be very potent antagonists for <I>r</I>TRPV1, blocking <I>in vivo</I> the hypothermic action of capsaicin, consistent with their <I>in vitro</I> mechanism. The docking study of compounds <B>13<I>S</I> </B> and <B>16<I>S</I> </B> in our <I>h</I>TRPV1 homology model indicated that the binding modes differed somewhat, with that of <B>13<I>S</I> </B> more closely resembling that of <B>GRT12360</B>.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

A Study on Single Dose Toxicity of Mecasin Pharmacopuncture Injection in Muscle

Eunhye Cha,Hohyun Jeong,Jongchul Lee,Seongjin Lee,Manyong Park,Sungchul Kim 대한한의학회 2015 대한한의학회지 Vol.36 No.2

A Single Residue Method for the Determination of Chlorpropham in Representative Crops Using High Performance Liquid Chromatography

Lee, Hyeri,Riu, Myoungjoo,Kim, Eunhye,Moon, Joon-Kwan,Choi, Hoon,Do, Jung-A,Oh, Jae-Ho,Kwon, Ki-Sung,Lee, Young Deuk,Kim, Jeong-Han 한국응용생명화학회 2013 Applied Biological Chemistry (Appl Biol Chem) Vol.56 No.2

A single residue analytical method was developed for herbicide chlorpropham in various crops. Brown rice, apple, mandarin, Kimchi cabbage, green pepper, potato, and soybean were selected as representative crops, and clean-up system, partition solvent, and extraction solvent were optimized. For high performance liquid chromatography (HPLC), $C_{18}$ column was used with elution solvents of water and acetonitrile. Limit of quantitation (LOQ) of chlorpropham was 2 ng (S/N >20), and excellent linearity ($R^2$=1.000) was achieved. Method limit of quantitation (MLOQ) was 0.02 mg/kg. For recovery tests, crop samples were macerated and fortified with chlorpropham standard solution at three fortification levels (MLOQ, 10 MLOQ, and 100 MLOQ). And then those were extracted with acetonitrile, concentrated and partitioned with n-hexane. The n-hexane layer was then concentrated, cleaned-up through Florisil$^{(R)}$ column with ethyl acetate:n-hexane (5:95, v/v) prior to concentration and analysis with HPLC. Good recoveries from 76.8 to 107.9% with coefficients of variation of less than 10% were obtained, regardless of sample type, which satisfies the criteria of Korea Food and Drug Administration. Those results were confirmed with liquid chromatography-mass spectrometry (LC-MS). The method established in this study could be applied to most of crops as an official and general method for the analysis of chlorpropham residue.

Exposure and Risk Assessment of Insecticide Methomyl for Applicator during Treatment on Apple Orchard

Eunhye Kim,문준관,Hoon Choi,Su-Myoung Hong,Dong-Hyuk Lee,Hyomin Lee,김정한 한국응용생명화학회 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.1

Exposure and risk assessments were conducted to evaluate safety of speed spayer (SS) and power sprayer (PS) used for treatment of insecticide methomyl in apple orchard on the operator. Dermal patches, gloves, socks, and masks were used to monitor the potential dermal exposure, and personal air monitor with XAD-2 resins was used to evaluate the potential inhalation exposure. Validation of methods for limit of detection, limit of quantitation, recovery, reproducibility, linearity of calibration, trapping efficiency, and breakthrough tests were performed to obtain reasonable results for quantitative exposure study of methomyl. During application of methomyl, PS resulted in more dermal exposure than SS. Important contaminated parts of body were upper arms, thigh, chest, shin, hand, forearm, and head for both SS and PS. Exposure rate was 44–176 mL/h. Although the level of inhalation exposure was very low during application, relatively higher level was observed for PS than for SS. During mixing/loading, more dermal exposure occurred by SS than that of PS probably due to drift of wettable powder (WP) formulation. Exposure was mostly observed on hand, and 99.9% of hand exposure to soluble liquid formulation (215 mg) in PS was from spill of liquid formulation on gloves. However, the body exposure ratio to total mixing/loading amount and inhalation exposure during mixing/loading was very low. Margin of safety in risk assessment was much larger than 1 in all cases, indicating low risk.

Rapid and Simultaneous Analysis of 360 Pesticides in Brown Rice, Spinach, Orange, and Potato Using Microbore GC-MS/MS

Lee, Jonghwa,Kim, Leesun,Shin, Yongho,Lee, Junghak,Lee, Jiho,Kim, Eunhye,Moon, Joon-Kwan,Kim, Jeong-Han American Chemical Society 2017 Journal of agricultural and food chemistry Vol.65 No.16

<P>A multiresidue method for the simultaneous and rapid analysis of 360 pesticides in representative agricultural produce (brown rice, orange, spinach, and potato) was developed using a modified QuEChERS procedure combined with gas chromatography tandem mass spectrometry (GC-MS/MS). Selected reaction monitoring transition parameters (e.g., collision energy, precursor and product ions) in MS/MS were optimized to achieve the best selectivity and sensitivity for a wide range of GC-amenable pesticides. A short (20 m) microbore (0.18 mm i.d.) column resulted in better signal-to-noise ratio with reduced analysis time than a conventional narrowbore column (30 m X 0.25 mm i.d.). The priming injection dramatically increased peak areas by masking effect on a new GC liner. The limit of quantitation was <0.01 mg/kg, and the correlation coefficients (r(2)) of matrix-matched standards were >0.99 within the range of 0.0025-0.1 mg/kg. Acetonitrile with 0.1% formic acid without additional buffer salts was used for pesticide extraction, whereas only primary secondary amine (PSA) was used for dispersive solid phase extraction (dSPE) cleanup, to achieve good recoveries for most of the target analytes. The recoveries ranged from 70 to 120% with relative standard deviations of <= 20% at 0.01 and 0.05 mg/kg spiking levels (n = 6) in all samples, indicating acceptable accuracy and precision of the method. Seventeen real samples from local markets were analyzed by using the optimized method, and 14 pesticides in 11 incurred samples were found at below the maximum residue limits.</P>