• Bulletin of the Korean Chemical Society
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• 제35권4호
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• pp.1133-1138
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• 2014

Ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI- MS/MS) provides a high-speed method to screen a large number of samples for small molecules with specific properties. In this study, UPLC-ESI-MS/MS with multiple reaction monitoring (MRM) was employed to screen urinary phospholipid (PL) content for biomarkers of prostate cancer. From lists of urinary PLs structurally identified using nanoflow LC-ESI-MS/MS, 52 PL species were selected for quantitative analysis in urine samples between 22 cancer-free urologic patients as controls and 45 prostate cancer patients. Statistical treatment of data by receiver operating characteristic (ROC) analysis yielded 14 PL species that differed significantly in relative concentrations (area under curve (AUC) > 0.8) between the two groups. Among PLs present at higher levels in prostate cancer urine, phosphatidylcholines (PCs) and phosphatidylinositols (PIs) constituted the major head group PLs (3 PCs and 7 PIs). For technical reasons, PL species of low abundance may be underrepresented in data from UPLC-ESI-MS/MS performed in MRM mode. However, the proposed method enables the rapid screening of large numbers of plasma or urine samples in the search for biomarkers of human disease.

• Annals of Clinical Neurophysiology
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• 제23권1호
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• pp.46-52
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• 2021

Background: We aimed to investigate candidates for serological biomarkers of neuropathic pain in individuals with neuromyelitis optica spectrum disorder (NMOSD). Methods: We analyzed 38 sera samples from 38 participants with NMOSD in National Cancer Center. Neuropathic pain was evaluated using the painDETECT questionnaire. Pain with neuropathic components (painDETECT score ≥ 13) was observed in 22 participants, among whom 17 had definite neuropathic pain (painDETECT score ≥ 19). The remaining 16 participants had non-neuropathic pain (painDETECT score < 13). Serum glial fibrillary acidic protein (GFAP) levels were assessed using a single-molecule array assay. Several cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-10, and IL-17A, were measured by a multiplex bead-based immunoassay. Results: In comparison of NMOSD participants with neuropathic pain components (or definite neuropathic pain) and those with non-neuropathic pain, the absolute values of serum GFAP, TNF-α, IL-6, and IL-10 levels were higher in participants with neuropathic pain components (or definite neuropathic pain), but these findings did not reach statistical significance. Conclusions: Further larger-scale investigations to find reliable serological biomarkers for neuropathic pain in NMOSD are warranted.

• Molecules and Cells
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• 제40권7호
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• pp.466-475
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• 2017

Dietary supplements have exhibited myriads of positive health effects on human health conditions and with the advent of new technological advances, including in the fields of proteomics, genomics, and metabolomics, biological and pharmacological activities of dietary supplements are being evaluated for their ameliorative effects in human ailments. Recent interests in understanding and discovering the molecular targets of phytochemical-gene-protein-metabolite dynamics resulted in discovery of a few protein signature candidates that could potentially be used to assess the effects of dietary supplements on human health. Persimmon (Diospyros kaki) is a folk medicine, commonly used as dietary supplement in China, Japan, and South Korea, owing to its different beneficial health effects including anti-diabetic implications. However, neither mechanism of action nor molecular biomarkers have been discovered that could either validate or be used to evaluate effects of persimmon on human health. In present study, Mass Spectrometry (MS)-based proteomic studies were accomplished to discover proteomic molecular signatures that could be used to understand therapeutic potentials of persimmon leaf extract (PLE) in diabetes amelioration. Saliva, serum, and urine samples were analyzed and we propose that salivary proteins can be used for evaluating treatment effectiveness and in improving patient compliance. The present discovery proteomics study demonstrates that salivary proteomic profile changes were found as a result of PLE treatment in prediabetic subjects that could specifically be used as potential protein signature candidates.

• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7575-7581
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• 2014

MicroRNAs (miRNAs) play an essential role in the development and progression of nasopharyngeal carcinomas (NPC). Despite advances in the field of cancer molecular biology and biomarker discovery, the development of clinically validated biomarkers for primary NPC has remained elusive. In this study, we investigated the expression and clinical significance of miRNAs as novel primary NPC diagnostic biomarkers. We used an array containing 2, 500 miRNAs to identify 22 significant miRNAs, and these candidate miRNAs were validated using 67 fresh NPC and 25 normal control tissues via quantitative real-time PCR (qRT-PCR). Expression and correlation analyses were performed with various statistical approaches, in addition to logistic regression and receiver operating characteristic curve analyses to evaluate diagnostic efficacy. qRT-PCR revealed five differentially expressed miRNAs (miR-93-5p, miR-135b-5p, miR-205-5p and miR-183-5p) in NPC tissue samples relative to control samples (p<0.05), with miR-135b-5p and miR-205-5p being of significant diagnostic value (p<0.01). Moreover, comparison of NPC patient clinicopathologic data revealed a negative correlation between miR-93-5p and miR-183-5p expression levels and lymph node status (p<0.05). These findings display an altered expression of many miRNAs in NPC tissues, thus providing information pertinent to pathophysiological and diagnostic research. Ultimately, miR-135b-5p and miR-205-5p may be implicated as novel NPC candidate biomarkers, while miR-93-5p, miR-650 and miR-183-5p may find application as relevant clinical pathology and diagnostic candidate biomarkers.

• 한국자원식물학회지
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• 제28권5호
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• pp.591-599
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• 2015

본 연구는 까마귀쪽나무 열매 70% 주정에탄올 추출물을 가지고 제작된 인체적용시험시료(LJTM)의 항염증 및 골관절염 biomarker의 변화를 통한 관절건강 기능성식품을 개발하기 위하여 인체적용시험시료를 제작한 후 이에 대한 유효성분의 함량을 평가하고, 인체적용시험전에 그 효력이 유지됨을 확인하기 위하여 수행되었다. 본 연구에 사용된 시료 LJTM은 NO생성이 억제되는 농도에서 세포독성이 관찰되지 않았으며, TNF-α와 IL-6 생성이 농도 의존적으로 억제되고 PGE₂를 억제하였다. 또한 동물시험에서 골관절염의 biomarker인 MMP-2, 3, 7, 9와 TIMP-1, 2에 대한 mRNA 발현이 농도 의존적으로 억제되었으며, 중추신경계 및 말초신경계에 의한 통증을 억제하는 것으로 평가되었다. 따라서 까마귀쪽나무 열매 70% 추출물이 함유된 인체적용시험시료(LJTM)는 골관절염과 진통억제에 우수한 효과가 있는 것으로 확인되었다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권4호
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• pp.1361-1365
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• 2015

Background: We aimed to discover potential gene biomarkers for gastric cancer (GC) diagnosis. Materials and Methods: Genechips of 10 GC tissues and 10 gastric mucosa (GM, para-carcinoma tissue, normal control) tissues were generated using an exon array of Affymetrix containing 30,000 genes. The differentially expressed genes (DEGs) between GC tissues and normal control were identified by the Limma package and analyzed by hierarchical clustering analysis. Gene ontology (GO) and pathway enrichment analyses were performed for investigating the functions of DEGs. Receiver operating characteristics (ROC) analysis was performed to measure the effects of biomarker candidates for diagnosis of GC. Results: Totals of 896 up-regulated and 60 down-regulated DEGs were identified to be differentially expressed between GC samples and normal control. Hierarchical clustering analysis showed that DEGs were highly differentially expressed and most DEGs were up-regulated. The most significantly enriched GO-BP term was revealed to be mitotic cell cycle and the most significantly enriched pathway was cell cycle. The intersection analysis showed that most significant DEGs were cyclin B1 (CCNB1) and cyclin B2 (CCNB2). The sensitivities and specificities of CCNB1 and CCNB2 were both high (p<0.0001). Areas under the ROC curve for CCNB1 and CCNB2 were both greater than 0.9 (p<0.0001). Conclusions: CCNB1 and CCNB2, which were involved in cell cycle, played significant roles in the progression and development of GC and these genes may be potential biomarkers for diagnosis and prognosis of GC.

• Genomics & Informatics
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• 제2권2호
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• pp.75-80
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• 2004

In a variety of animal species, the perinatal exposure of experimental animals to the 2,3,7,8-tetrachlorodibenzo­p-dioxin (TCDD) leads to the immune dysfunction, which is more severe and persistent than that caused by adult exposure. We report here the changes of gene expression and the identification of the marker-genes representing the dioxin exposure. The expressions of the transcripts were analyzed using the 11 K oligonucleotide­microarray from the bone marrow cells of male C57BL/6J mice after an intraperitoneal injection of $1{\mu}g$ TCDD/kg body weight at various time intervals: gestational 6.5 day(G6.5), 13.5 day(G13.5), 18.5 day(G18.5), and postnatal 3 (P3W)and 6 week (P6W). The type of self-organizing maps(SOM) representing the specific exposure dioxin could be identified as follows; G6.5D(C14), G13.5D(C0, C5, C10, C18), G18.5D(7): P3W(C2, C21), and P6W(C4, C15, C20). The candidate marker-genes were restricted to the transcripts, which could be consistently expressed greater than $\pm$2-fold in three experiments. The resulting candidates were 85 genes, the characteristics of that were involved in cell physiology and cell functions such as cell proliferation and immune function. We identified the biomarker-genes for dioxin exposure: smc -like 2 from SOM C14 for the dioxin exposure at G6.5D, focal adhesion kinase and 6 other genes from C0, and protein tyrosine phosphatase 4a2 and 3 other genes from C5 for G13.5D, platelet factor 4 from C7 for G18.5D, fos from C2 for P3W.

• Journal of Ginseng Research
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• 제45권2호
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• pp.273-286
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• 2021

Background: Prostate carcinoma is the second most common cancer among men worldwide. Developing new therapeutic approaches and diagnostic biomarkers for prostate cancer (PC) is a significant need. The Chinese herbal medicine Panax quinquefolius saponins (PQS) have been reported to show anti-tumor effects. We hypothesized that PQS exhibits anti-cancer activity in human PC cells and we aimed to search for novel biomarkers allowing early diagnosis of PC. Methods: We used the human PC cell line DU145 and the prostate epithelial cell line PNT2 to perform cell viability assays, flow cytometric analysis of the cell cycle, and FACS-based apoptosis assays. Microarray-based gene expression analysis was used to display specific gene expression patterns and to search for novel biomarkers. Western blot and quantitative real-time PCR were performed to demonstrate the expression levels of multiple cancer-related genes. Results: Our data showed that PQS inhibited the viability of DU145 cells and induced cell cycle arrest at the G1 phase. A significant decrease in DU145 cell invasion and migration were observed after 24 h treatment by PQS. PQS up-regulated the expression levels of p21, p53, TMEM79, ACOXL, ETV5, and SPINT1 while it down-regulated the expression levels of bcl2, STAT3, FANCD2, DRD2, and TMPRSS2. Conclusion: PQS promoted cells apoptosis and inhibited the proliferation of DU145 cells, which suggests that PQS may be effective for treating PC. TMEM79 and ACOXL were expressed significantly higher in PNT2 than in DU145 cells and could be novel biomarker candidates for PC diagnosis.

• 한국산학기술학회논문지
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• 제14권8호
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• pp.4086-4092
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• 2013

Mycosporine-like 아미노산(MAAs)은 UV 흡수물질이며, 다양한 해양생물들은 MAAs의 합성과 축적을 통하여 환경 자외선의 직 간접적인 영향을 감소시키는 기능을 진화시켜 왔다. 이 연구에서 우리는 radiofrequency(RF) 발생장치를 제작하였고, 이를 미세조류 배양에 적용하였다. $0.35{\pm}0.05$ mHz의 RF를 Chlamydomonas sp. 배양기에 공급하였고, 정해진 시간(0, 0.5, 1 및 2 시간)에 시료를 채취하였다. MAAs 생합성 관련 유전자인 dehydroquinate synthase homolog (DHQS-like)와 nonribosomal peptide synthetase homolog (NRPS-like) 유전자를 Chlamydomonas sp.로부터 클로닝하였고, RF 노출에 대한 유전자 발현을 qRT-PCR을 이용하여 분석하였다. 연구결과 RF에 노출된 Chlamydomonas sp.의 DHQS-like와 NRPS-like 유전자의 발현은 노출 1시간에 각각 1.46 배 및 1.19 배 증가하였다. 이러한 결과는 DHQS-like와 NRPS-like 유전자가 Chlaydomonas sp.의 MAAs 생합성을 진단할 수 있는 좋은 바이오마커 후보가 될 수 있음을 의미한다.

• 한국식품조리과학회지
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• 제15권2호
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• pp.108-113
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• 1999

참마(Dioscorea japonica Thunb.), 달개비(Commelina communis L.) 전초, 하늘타리(Trichosanthes kiri1owii Max.) 및 둥굴레(Polygonatum odoratum var. Plurifloum Ohwi) 뿌리의 MeOH 추출물과 petroleum ether/diethyl ether 및 EtOAc 가용성 용매분획물을 인체 폐암 및 난소선암세포주에 적용하여 암세포주 성장율을 측정하였다. 실험 결과 하늘타리 뿌리의 MeOH 추출물이 인체 폐암 및 난소선암세포주에 대해 모두 강한 성장억제를 보여주었으며 40$\mu\textrm{g}$/ml의 농도에서 30% 이하의 성장율을 나타내었다. 다음으로 용매분획물에 대한 세포독성은 하늘타리의 EtOAc 가용성 분획물에서 폐암과 난소선암세포주에 대한 성장억제율이 30% 이하로 매우 높게 나타나고 있으며, 참마의 EtOAc 가용성 용매분획물이 인체 폐암세포주에 대하여 30% 이하의 높은 성장억제를 나타내고 있다. 그밖에 달개비 전초의 EtOAc 가용성 분획물이 인체 폐암세포주에 대해 30%에서 60%의 성장율을 나타내고, 둥굴레 뿌리의 petroleum ether/diethyl ether 및 EtOAc 분획물이 인체 난소선암세포주에 대해 동일한 정도의 성장율을 보여주고 있다. 이상의 결과를 통해 한국산 야생식용식물로 부터 세포 독성작용을 갖는 새로운 식품 개발의 가능성을 확인할 수 있었을 뿐 아니라, 이를 위한 지속적인 연구로서 폐암 및 난소선암세포주에 대한 세포독성 작용이 가장 강하게 나타난 하늘타리 및 참마의 EtOAc 용매 세분획물을 이용하여 활성이 높은 순수 물질을 분리하므로 추후 새로운 항암활성 물질 연구를 위한 후보식물로서의 개발가능성을 보여주었다.