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개선된 플라스미드 DNA 전달 효율을 위한 히알루론 아민 코팅 폴리에틸렌이민 기반 전달 시스템
오경연 ( Kyoung-yeon Oh ),장용호 ( Yongho Jang ),이은비 ( Eunbi Lee ),김태호 ( Tae-ho Kim ),김현철 ( Hyuncheol Kim ) 한국공업화학회 2022 공업화학 Vol.33 No.1
현재 진행 중인 코로나19의 세계적 유행을 기점으로 유전자 전달을 통한 면역 형성에 대한 연구가 활발히 진행되고 있다. 특히 바이러스를 통한 유전자 전달이 부작용이 다수 발견됨에 따라 비바이러스성 유전자 전달체에 대한 요구가 크게 증가하였다. 본 연구에서는 생체적합물질인 히알루론 아민으로 코팅한 폴리에틸렌이민-플라스미드 DNA 복합체를 통한 효율적인 유전자 전달 시스템을 제안했다. 다양한 조성에서 생성된 폴리에틸렌이민-플라스미드 DNA 복합체(polyplex)와 히알루론 아민으로 코팅한 폴리에틸렌이민-플라스미드 DNA 복합체(polyplex-HA)의 크기 및 플라스미드 DNA 발현 정도를 비교해 각 물질의 최적 비율을 찾아냈고 복합체의 크기 및 제타 전위, 에너지 필터링 투과 전자현미경(EF-TEM) 이미지를 통해 입자의 특성을 평가했다. 세포 내 전달 및 발현 효율을 형광현미경과 유세포분석기를 통해 상용화 되어있는 유전자 전달체인 lipofectamine과 비교 분석했다. 본 연구에서 제안된 polyplex-HA는 pDNA 뿐만 아니라 다양한 유전물질을 전달할 수 있으며, 전달체에 대한 면역반응이 적어 다회성 투여에 유리하여 미래의 백신플랫폼의 기반이 될 수 있을 것으로 기대할 수 있다. Since the pandemic of COVID-19, active investigation to develop immunity to infectious disease by delivering nucleic acids has been proceeded. Particularly, many studies have been conducted on non-viral vector as several vital side-effects which were found on nucleic acid delivery system using viral vectors. In this study, we have developed plasmid DNA (pDNA) loaded- hyaluronic acid derivative (HA) coated-polyethyleneimine (PEI) based polyplex for enhanced nucleic acid delivery efficiency. We have optimized the ratio of pDNA : PEI : HA by measuring size and protein transcription efficiency. The final product, polyplex-HA, was characterized through measuring size, zeta-potential and TEM image. Intracellular uptake and protein transcription efficiency were compared to commercially available transfection reagent, lipofectamine, through fluorescence image and flow cytometry. In conclusion, polyplex-HA presents a novel gene delivery system for efficient and stable protein transcription since it is available for delivering various genetic materials and has less immunoreactivity.
한호연,오경의 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.4
Acanthiophilus helianthi (Rossi) (safflower fly) is known to host a wide range of plant species belonging to the tribe Cardueae (Asteraceae). It is also known as a serious pest of safflower in the dry regions and marginal areas of the world where safflower is an important oilseed crop. Due to its possible introduction to Korean peninsula, it is listed as a managed quarantine pest by the Korean Animal and Plant Quarantine Agency. We here report a natural occurrence of A. helianthi for the first time in Korea, based on a single female specimen collected from northern part of South Korea. Since the discovery of A. helianthi is of agricultural and quarantine importance in this part of world, we here provide its detailed morphological redescription, diagnosis, and color photographs including genitalic structures for accurate identification. We also provide a brief account of its biology and pest status summarized from previous literature. In order to make sure the correct identification of the newly discovered Korean specimen of A. helianthi as well as to investigate systematic position of the genus Acanthiophilus, we conducted a DNA barcoding analysis. Finally, we here suggest that A. helianthi should be removed from the Korean quarantine pest list, since its natural occurrence in Korea is confirmed by the present study.
Photodynamic Therapy for the Treatment of Pancreatic Cancer Using Chlorin e6 loaded Exosome
정재훈,장용호,오경연,김민균,김현철 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-
Exosome, extracellular vesicles for cell-to-cell communications, has been utilized as drug delivery system. In this study, tumor-derived exosome was sonicated and reassembled for a drug delivery carrier. Chlorin e6 (Ce6), a representative photosensitizer, was loaded into tumor-derived exosome during re-assembly of exosomes. The reassembled exosomes was found to have membrane proteins like the exosomes. By removing the internal substances, cancer formation from tumor-derived exosomes could be resolved. In vitro experimental data showed that Ce6-loaded re-assembled exosomes were selectively targeted to original-derived cells. Due to the high specific targeting ability of exosome, the Ce6 could be effectively delivered. Furthermore, after effective intracellular delivery, sufficient reactive oxygen species (ROS) were generated with very small amounts of Ce6 in the target cells under the laser irradiation. In conclusion, photodynamic therapy could be obtained with Ce6- loaded exosome.
초등학생의 골성숙도에 따른 체력 군집화 : 군집분석 중심으로
김대훈,윤형기,오세이,이영준,조석연,송대식,서동녘,김주원,나규민,김민준,오경아 한국응용과학기술학회 2022 한국응용과학기술학회지 Vol.39 No.1
The aim of this study was to cluster according to the bone age of elementary school students in order to analyze the physique, physical fitness, and skeletal maturation of each cluter group and to provide basic data for the balanced development of elementary school students through data analysis. The subjects of this study were 2243 students aged 8 to 13 years, and the skeletal maturation were calculated by applying them to the TW3 method score conversion table after the X-ray films were taken. A total of 2 components in physique were measured using a stadiometer(Hanebio, Korea, 2021) and the Inbody 270(Biospace, Korea, 2019), and a total of 7 components in physical fitness, which included muscular strength(Hand Grip Strength), balance(Bass Stick Test), agility(Plate Tapping), power(Standing Long Jump), flexibility(Sit&Reach), muscular endurance(Sit-Up), and cardiovascular endurance(Shuttle Run) were measured as well. K-Means clustering method, cross-tabulation analysis, and one-way variable analysis(ANOVA) were conducted for data processing using the SPSS PC/Program(Version 26.0) and Bristics Studio Tool, and it was considered significant at the level of p< .05. The results of this study may be summarized as follow. First, as a result of clustering using three components of skeletal maturation: retarded, normal, and advanced, cluster 1(Retarded) showed excellence in muscular strength, balance, and agility. cluster 2(Normal) showed poor flexibility, whereas cluster 3(Advanced) showed excellence in muscular strength. Second, as a result of analyzing the differences in physique according to the clustering of elementary school students by their individual characteristics, cluster 3(Advanced) showed excellence in height, weight, and body fat percentage. Third, as a result of analyzing the differences in physical fitness according to the clustering of elementary school students by their individual characteristics, cluster 3(Advanced) showed excellence in Hand Grip Strength(Left, Right), whereas cluster 1(Retarded) showed excellence in Bass Stick Test, and cluster 3(Advanced) showed excellence in Standing Long Jump. 본 연구는 초등학생의 골연령에 따라 군집화 시켜 각 군집 그룹의 체격, 체력 및 골성숙도를 분석하고 자료 분석을 통해 초등학생들의 균형적인 발달을 위한 기초자료를 제공하는 데 있다. 연구대상은 8세∼13세에 해당하는 2243명을 대상으로 하였으며 골성숙도 산출을 위해 X-ray필름을 촬영한 후 TW3 방법 점수 환산표에 적용시켜 골성숙도를 산출했다. 신장계(Hanebio, Korea, 2021)와 Inbody 270 (Biospace, Korea, 2019)를 사용하여 총 2개의 체격 요소를 측정하였으며, 체력은 근력(악력), 평형성(외발 서기), 민첩성(플랫테핑), 순발력(제자리멀리뛰기), 유연성(좌전굴), 근지구력(윗몸일으키기), 심폐지구력(셔 틀런)으로 총 7개 체력 요소의 종목을 측정하였다. 자료처리 방법은 SPSS PC/Program(Version 26.0)과 Britics Studio Tool을 이용하여 K-Means 클러스터링 기법, 교차분석, 일원변량분석(One-Way ANOVA) 을 실시하였으며, p< .05 수준에서 유의한 것으로 간주하였다. 본 연구의 결과는 다음과 같다. 첫째, 미숙, 보통, 조숙의 3가지 골성숙도를 사용하여 군집화한 결과, 군집 1(미숙)은 근력, 평형성, 민첩성에서 높게 나타났다. 군집 2(보통)는 유연성에서 낮게 나타났으며, 군집 3(조숙)은 근력에서 높게 나타났다. 둘째, 초등 학생의 개인특성별 군집화에 따른 체격 차이를 분석한 결과, 신장, 체중, 체지방률 모두 군집 3(조숙)이 높게 나타났다. 셋째, 초등학생의 개인특성별 군집화에 따른 체력 차이를 분석한 결과, 악력검사(좌, 우)는 군집 3(조숙)이 높게 나타났고 외발서기의 경우 군집 1(미숙)이 높게 나타났으며, 제자리멀리뛰기의 경우 군집 3(조숙)이 높게 나타났다.
Inhibition of ATR Increases the Sensitivity to WEE1 Inhibitor in Biliary Tract Cancer
남아롱,JINMEIHUA,방주희,오경석,서혜림,오도연,방영주 대한암학회 2020 Cancer Research and Treatment Vol.52 No.3
Purpose Currently, the DNA damage response (DDR) pathway represents a key target for new cancer drug development. Advanced biliary tract cancer (BTC) has a poor prognosis because of the lack of efficacious treatment options. Although DNA repair pathway alterations have been reported in many patients with BTC, little is known regarding the effects of DDR-targeted agents against BTC. Materials and Methods In this study, nine BTC cell lines were exposed to the WEE1 inhibitor (AZD1775). In vitro, MTT assay, colony-forming assay, cell cycle analysis, phospho-histone H3 staining assay, Transwell migration assay, and western blot were performed. Then, to enhance the antitumor effect of AZD1775, the combination treatment of WEE1 inhibitor and ataxia telangiectasia mutated and Rad3 related (ATR) inhibitor (AZD6738) was conducted using MTT assay and comet assay. Finally, HuCCT-1 and SNU2670 xenograft models were established to confirm the anti-tumor effect of AZD1775 alone. Furthermore, the combination treatment was also evaluated in SNU2670 xenograft models. Results AZD1775 blocked the phosphorylation of CDC2 and CDC25C in all cell lines, but significantly increased apoptosis and S phase arrest in sensitive cells. However, increased p-ATR and phosphorylated ataxia telangiectasia mutated levels were observed in less sensitive cells. In addition, in vitro and in vivo data illustrated that AZD1775 combined with AZD6738 exerted more potent anti-tumor effects than either drug alone. Although WEE1 inhibition has promising anti-tumor effects in some BTC cells, the addition of ATR inhibitors could enhance its efficacy. Conclusion Taken together, this study supports further clinical development of DDR-targeted strategies as monotherapy or combination regimens for BTC.
Inhibition of WEE1 Potentiates Sensitivity to PARP Inhibitor in Biliary Tract Cancer
서혜림,남아롱,방주희,오경석,김재민,윤지선,김태용,오도연 대한암학회 2022 Cancer Research and Treatment Vol.54 No.2
Purpose Up to 20% of patients with biliary tract cancer (BTC) have alterations in DNA damage response (DDR) genes, including homologous recombination (HR) genes. Therefore, the DDR pathway could be a promising target for new drug development in BTC. We aim to investigate the anti-tumor effects using poly(ADP-ribose) polymerase (PARP) and WEE1 inhibitors in BTC. Materials and Methods We used 10 BTC cell lines to evaluate an anti-tumor effect of olaparib (a PARP inhibitor) and AZD1775 (a WEE1 inhibitor) in in vitro. Additionally, we established SNU869 xenograft model for in vivo experiments. Results In this study, we observed a modest anti-proliferative effect of olaparib. DNA double-strand break (DSB) and apoptosis were increased by olaparib in BTC cells. However, olaparib-induced DNA DSB was repaired through the HR pathway, and G2 arrest was induced to secure the time for repair. As AZD1775 typically regulates the G2/M checkpoint, we combined olaparib with AZD1775 to abrogate G2 arrest. We observed that AZD1775 downregulated p-CDK1, a G2/M cell cycle checkpoint protein, and induced early mitotic entry. AZD1775 also decreased CtIP and RAD51 expression and disrupted HR repair. In xenograft model, olaparib plus AZD1775 treatment reduced tumor growth more potently than did monotherapy with either drug. Conclusion This is the first study to suggest that olaparib combined with AZD1775 can induce synergistic anti-tumor effects against BTC. Combination therapy that blocks dual PARP and WEE1 has the potential to be further clinically developed for BTC patients. Purpose Up to 20% of patients with biliary tract cancer (BTC) have alterations in DNA damage response (DDR) genes, including homologous recombination (HR) genes. Therefore, the DDR pathway could be a promising target for new drug development in BTC. We aim to investigate the anti-tumor effects using poly(ADP-ribose) polymerase (PARP) and WEE1 inhibitors in BTC.Materials and Methods We used 10 BTC cell lines to evaluate an anti-tumor effect of olaparib (a PARP inhibitor) and AZD1775 (a WEE1 inhibitor) in in vitro. Additionally, we established SNU869 xenograft model for in vivo experiments.Results In this study, we observed a modest anti-proliferative effect of olaparib. DNA double-strand break (DSB) and apoptosis were increased by olaparib in BTC cells. However, olaparib-induced DNA DSB was repaired through the HR pathway, and G2 arrest was induced to secure the time for repair. As AZD1775 typically regulates the G2/M checkpoint, we combined olaparib with AZD1775 to abrogate G2 arrest. We observed that AZD1775 downregulated p-CDK1, a G2/M cell cycle checkpoint protein, and induced early mitotic entry. AZD1775 also decreased CtIP and RAD51 expression and disrupted HR repair. In xenograft model, olaparib plus AZD1775 treatment reduced tumor growth more potently than did monotherapy with either drug.Conclusion This is the first study to suggest that olaparib combined with AZD1775 can induce synergistic anti-tumor effects against BTC. Combination therapy that blocks dual PARP and WEE1 has the potential to be further clinically developed for BTC patients.