Selective Regulation of Micro-RNAs in Breast Cancer

손유경 전주대학교 대학원 2017 국내석사

The effect of plant-derived nanovesicles on cancer metastasis in 3D microfluidic model : Anti-cancer effect of plant sap-derived nanovesicles

김기민 국제뇌교육종합대학원대학교 2018 국내석사

Natural plants are valuable drug candidates as they have been widely used as traditional therapeutics in Asia. The aim of this paper is to investigate the cytotoxic effects of the nanovesicles derived from four endemic plants: Dendropanax morbifera (DM), Pinus densiflora (PD), Chamaecyparis obtusa (CO) and Platycladus orientalis (PO) against cancerous cells. The crux of this research is to compare the cytotoxicity of the nanovesicles between malignant and benign tumor cells as this is a key factor in determining the targeting ability of the nanovesicles treatment to possibly replace chemotherapy that has detrimental side effects on normal healthy body cells. The results show that nanovesicles from DM and PD are promising as anti-cancer drug candidates while the others present less or no cytotoxicity to tumor cells. Mixing DM and PD derived nanovesicles together greatly increased their targeting ability against malignant breast and skin cancer cells. Then, we investigated the role of the sap in anti-metastatic effects using a cancer metastasis model based on 3D microfluidic system which closely mimics in vivo tumor environment. We found that the nano-sized vesicles of the sap were effective in suppression of cancer metastasis. This 3D microfluidic model could be widely used for various drug screening assays validating therapeutic effect on cancer metastasis and can be alternatives to animal testing.

나노 영가철을 이용한 국내 지하수내 질산성질소(NO3--N) 소거 활용성 조사

유선일 전주대학교 일반대학원 2017 국내석사

As a result of the survey on major domestic water pollutants in the recent five years, the average of total samples exceeding the guideline was 5.2%, among which NO3--N(Nitrate-Nitrogen) was 22.1%. The most commonly used removal method for the NO3--N was ion exchange resin so far. The research trend of the nZVI(nano-Zero Valent Iron) was conducted through keyword search during the last 10 years. The number of foreign documents was 1,410 and showed an increasing trend in every year. However, there were 33 domestic documents and difficult to find the increasing tendency. Typically, the nZVI is synthesisd through the consequence processes of mixing step, separation step, cleaning step, and drying step. However, the nZVI is so quickly oxidative, that corrosion and coagulation are happened together. Recently, additional methods such as surface modification, immobilization, dual solvent, and others have been applied to synthesis of the nZVI. In this study, the ability of nZVI prepared without washing process was compared with nZVI prepared by clean up process, including the GT-nZVI(Green Tea-nZVI) synthesized utilizing green tea. The NO3--N concentration was measured by using spectrophotometric method and the removal efficiency of NO3--N was ranged between 16% and 79%, depending on the experimental methods. The nZVI was used to remove heavy metals and ionic pollutants, especially NO3--N removal. Research has also shown that experiments involving the removal of NO3--N accounted for a large proportion. From this point of view, the nZVI will be expected to us as one of the practical ways in the future for removal of NO3--N rather than other pollutants. The ability of the nZVI to remove contaminants, synthesis and removal mechanisms of the literature are suggested. The comparison studies with existing methods were investigated for removing NO3--N from the ground water. As a result, the bio-degradable polymer of the surface modification of the nZVI was found to be the most practical method in the field. It is expected that surface deformation studies and trials for testing practical use of the nZVI will be worthy to pursue further. Keywords: nZVI, Green Tea, Nitrate-Nitrogen, Polymer, Surface modific- ation, mechanism, ground water

Selective Regulation of Epithelial-Mesenchymal Transition (EMT) Using Micro-RNAs in Breast Disease

김민지 전주대학교 대학원 2017 국내석사

Most of breast cancer (BC) metastasis is caused by epithelial-mesenchymal transition (EMT) that invades other organs by changing their characteristics. A new research is required for real-time information, detection of EMT processes, and better treatment with focusing. As a small non-coding RNA molecules, microRNA (miRNA) is to degrade the target mRNA by destabilizing and suppressing the further processes of translation. Because this miRNA can bind binds through seed sites which have 2 to 8 nucleotides. Thus, one gene can be regulated by a variety of miRNAs. It is necessary to study the effect of miRNAs as new molecular mechanistic modulators in tumor progression and inflammation. In this study, I tried to find common miRNAs which are linked with the several tumor suppressor genes using big data mining and validation. I identified that miR-93-5p and miR-221-3p are associated with inflammation of breast epithelial cells. Induction of EMT in mammary epithelial cell (MCF-10A) was triggered by transforming growth factor beta (TGF-β1) treatment. In TGF-β-induced EMT processes, miR-93-5p and miR-221-3p were double checked as a common miRNAs which are elevated and linked to the degradation of five tumor suppressor genes. MiR-93-5p and miR-221-3p overexpression levels were elucidated by real-time PCR. When I verified the protein expression level of EMT markers including E-cadherin, N-cadherin, and fibronectin, the expression level of N-cadherin and fibronectin were significantly increased in EMT- induced MCF-10A. However, the level of E-cadherin was significantly reduced vice versa. Likewise, expression level of tumor suppressor gene Smad4 is significantly increased in EMT- induced MCF-10A. After that when I transfected with miR-93-5p and miR-221-3p anti-sense oligonucleotide (ASO), the reduced tumor suppressor Smad4 protein expression was recovered. Through this study, I can conclude that the data mined miRNA candidates which can regulate the five tumor suppressor genes were validation as a common miRNA in EMT-induced breast cancer progression. Keywords: microRNA, big data mining, epithelial to mesenchymal transition (EMT), tumor suppressor gene, breast cancer prevention.

Selective regulation of micro-RNAs in breast cancer

손유경 전주대학교 일반대학원 2011 국내석사

Breast cancer classified in the presence and/or absence of estrogen receptor (ER), progesterone receptor (PR) and human EGF-like receptor type 2 (HER2). Triple negative breast cancer (TNBC) is a one kind of breast cancers that ER, PR, HER2 are totally negative. Breast cancer must be taken care of consistently because breast cancer often has a relapse frequently in 2 to 3 years after surgery include TNBC. It is necessary to elucidate the early detection molecular makers. Micro-RNAs (miRNAs) are approximately 22-nucleotide non-coding RNAs and play an important role in inhibition of translation or degradation of targeted messenger RNA (mRNA). A number of miRNAs were found to have oncogenic potential and tumor suppressors potential as well. Up to date, many studies of miRNA focus on only one or two single miRNA targets with one gene. However, miRNA’s multi-targeting ability to genes is barely carried out. By utilizing big data mining, I found the positive correlation between miRNAs and tumor suppressor genes such as PTEN, Smad4, Rb1, BRCA1, BRCA2. To validate the findings by big data mining, I could check the overexpressed miR-93-5p and miR-106a-5p are linked to the tumor suppressor genes regulations in human breast cancer cell lines MCF-7, MDA-MB-231 by real time-PCR. Moreover, transfection anti-sense oligonucleotide (ASO) of miR-93-5p and miR-106a-5p lead to increase of tumor suppressor PTEN protein levels in human breast v cancer cell lines MCF-7 and MDA-MB-231. Inversely, transfection ASO of miR-93-5p and miR-106a-5p lead to decrease the tumor suppressor PTEN protein levels in human breast normal cell lines MCF-10A. As a result, miR-93-5p and miR-106a-5p can regulate tumor suppressor PTEN. In conclusion, these results support that modulation of elevated miR-93-5p and miR-106a-5p can help in breast cancer prevention via regulate tumor suppressor PTEN. Keywords: micro-RNA, big data mining, target genes, breast cancer prevention

엑소좀에 의한 암-연관 섬유아세포 유도와 암세포의 침윤성 증가에 대한 연구

손예주 국제뇌교육종합대학원대학교 2019 국내석사

암이란 정상세포의 유전적인 돌연변이 및 결함, 환경적인 요인 등 여러 가지 복합적인 요인으로 인하여 세포가 비정상적으로 성장, 분열하여 정상 조직에도 생체 기능 이상이 생기는 것을 말한다. 특히 암 전이로 인한 사망률은 90%에 달하여 암 전이의 억제방법에 대한 연구의 필요성이 대두되고 있다. 또한, 최근에는 암세포 자체보다는 종양미세환경의 중요성을 인지하고 이에 대한 연구가 집중되고 있다. 본 연구는 미세유체소자를 이용해 생체 외에서 종양미세환경을 구현하고, 암세포의 이동 및 침윤성에 영향을 주는 다양한 인자들 중 암-연관 섬유아세포의 가치를 밝히고자 하였다. 더불어 암-연관 섬유아세포의 유도에 중요한 역할을 하는 암세포 유래 엑소좀과의 관계를 살펴보고, 암-연관 섬유아세포로 인한 암세포의 이동 및 침윤성의 관계를 분석하였다. 엑소좀은 암세포가 분비하는 인자로서 mRNA, miRNA, 단백질 등 유전 정보를 담고 있고, 입자가 작아서 다른 세포에 유입되기 쉬워 정상세포를 다른 세포로 분화시키기도 한다. 이렇게 정상세포가 분화되어 암-연관 섬유아세포가 되어 주변 조직의 단백질을 분해하여 물리적인 틈을 만들어 암세포의 이동 및 침윤에 영향을 준다. 이것을 3차원 미세유체소자를 이용해 전이와 관련이 있는 유방암, 악성 흑색종, 피부암의 암세포를 이용해 실험을 하였다. 각 암세포에서 엑소좀을 분리하고 분리한 엑소좀으로 인해 혈관내피세포가 암-연관 섬유아세포로 분화되는 것을 실시간 촬영하여 확인하고, 각 암세포별로 유도된 암-연관 섬유아세포의 개수, 면적, 거리를 비교해보았다. 또한 암-연관 섬유아세포가 암세포의 침윤성에 어떤 영향을 끼치는지 알아보기 위해 혈관에 암-연관 섬유아세포가 있는 경우와 그렇지 않은 경우에 암세포가 세포외 기질로 침윤하는 정도를 실시간으로 촬영하였다. 그리고 촬영한 이미지를 이용해 침윤한 암세포의 개수, 면적, 거리를 비교해보았다. 실험 결과, 암세포만 있을 때보다 암-연관 섬유아세포가 있을 때 침윤한 암세포의 개수가 많았다, 또한 암-연관 섬유아세포가 있을 때 침윤한 면적이 더 크고, 더 먼 거리로 침윤한다는 것으로 밝혀졌다. 이것을 통해 암-연관 섬유아세포가 암세포를 물리적인 방법으로 다른 장기로 침윤을 유도한다는 것을 알 수 있었다. 따라서 암 전이에 있어서 암-연관 섬유아세포가 중요한 역할을 하며, 암을 진단할 수 있는 바이오 마커로서 긍정적인 효과를 기대해 볼 수 있다. 그리고 이 플랫폼은 암-연관 섬유아세포를 억제하는 치료제 개발에 유용할 것이다.

다양한 기능성 소재가 피부에 미치는 영향에 대한 국내 문헌 연구

김종현 전주대학교 일반대학원 2017 국내석사

In the 21st century, people are getting more interested in skin care. Related industries and researches are actively being performed. Due to the lack of necessary nutrients and vitamins, bad eating habits, and excess fat intake, people are suffering from skin problems caused by internal health issues. The skin is wrapped around the outermost part of the human body and is continuously exposed to the outside conditions. It is stimulated by exposure to various environmental factors, stresses and ultraviolet rays, which proceed of exogenous aging. In addition, synthesis and accumulation of melanin bring problems such as erythema, age spots, freckles and dark spots. Recently, markets of functional cosmetics and health food are growing rapidly as demand for easier and more practical way to rejuvenate the skin health. Diverse products for pursuing this aim are also released accordingly. This study investigated the literature regarding the functional substances to keep and or rejuvenate skin health. For whitening by inhibition of melanin and tyrosinase, for removing wrinkles by inhibition of elastase and collagenase, and for protecting ultraviolet rays, multiple compelling experiments were analyzed. Domestic literature collected from 2007 to 2017 has been reported 66 functional substances and their molecular mechanisms. International literature searched from 2015 to 2017 has been reported 32 functional substances to help and keep skin health. Keyword : Whitening, Wrinkle, Againg, Melanin, Tyrosinase, Collagen,

홍삼 유래물질이 희소돌기아교세포의 분화와 막형성에 미치는 영향

정귀련 국제뇌교육종합대학원대학교 2021 국내석사

Ginseng is a plant that used a lot of medicinal roots. The use of saponin, which is a major pharmacological component, contains nucleic acids, alkaloids, and vitamins, and uses enhancement, fatigue recovery, control of functions such as the nervous and metabolic systems. Saponin is classified as a glycoside called ginsenoside, and in red ginseng made by steaming ginseng, it is present in the ginsenoside and in the gin senoside. Since some ginsenosides have effective effects on the central nervous system, in this paper, we will use ginsenosides in oligodendrocytes that generate myelin sheaths to determine whether red ginseng-derived substances are treated with rare stones. Red ginseng extract and its main components of red ginseng, saponin and non-saponin, and then the major ginsenosides contained in saponin, were tested on oligodendrocytes. We would like to analyze how it affects it to see if its effectiveness is unique. As a result of the study, although saponin can induce membrane formation well, there was a significant difference according to the concentration difference. 11 ginsenosieds, among the saponins, Rb1 induces better differentiation compared to DMSO. Ginsenosides Rb1 is substances that have good efficacy on nerve cells and central nervous system, and have a positive effect on the formation of oligodendrocytes from differentiation. The most significant effect of each substance is further studied, such as the optimal concentration, and the items that ginsenosides have a significant advantage in the differentiation of oligodendrocytes are meaningful. Key Words : Red ginseng, saponin, ginsenoside, myelin, oligodendrocyte 인삼은 뿌리를 약용으로 많이 사용하던 식물이다. 주요 약리 성분인 사포닌을 비롯하여 아미노산, 핵산, 알칼로이드, 비타민들이 함유되어있으며, 체력 증강, 피로 회복, 신경계, 대사계 등의 기능 조절 등을 하는데 사용되었다. 사포닌은 진세노사이드라는 배당체로 분류되는데 인삼을 증숙하여 만든 홍삼에는 인삼의 진세노사이드에 더해, 홍삼에는 존재하지 않는 진세노사이드들도 존재한다. 진세노사이드에는 중추신경계에 대한 유효한 효능을 가진 성분들이 많이 있으므로 본 논문에서는 수초를 생성하는 희소돌기아교세포에 진세노사이드를 비롯한 홍삼유래 물질을 처리하여, 물질들이 희소돌기아교세포에 어떠한 영향을 가지는 지를 보고자 하였다. 홍삼 진액과 그 홍삼의 주요 성분인 Saponin과 non-saponin을, 그 다음으로 saponin에 포함되어있는 주요 ginsenosides를 희소돌기아교세포에 시험하여 희소돌기아교세포의 분화상태에서 세포수, 분화, 막형성에 어떠한 영향을 가지는 지를 분석하여 유효성을 가지는 지를 알아보고자 하였다. 연구 결과, 사포닌이 막형성은 잘 유도하였으나, 농도 차에 따른 분석에서는 유의한 차이가 나타나지 않았다. 사포닌 중에서도 진세노사이드 11종 간에서는 Rb1이 분화를 control인 DMSO에 비해 더 잘 유도하는 것으로 나타났다. 진세노사이드 Rb1은 신경세포와 중추신경계에 대한 효능이 알려져 있는 물질들로 희소돌기아교세포의 분화와 막형성을 잘 유도하였다는 것은 Rb1이 희소돌기아교세포의 수초의 형성에도 긍정적인 영향을 가질 것이라고 볼 수 있다. 각 물질이 in vivo에서도 유의한 효과를 나타내는지 추가적으로 연구를 하여야하지만, 사포닌과 진세노사이드가 희소돌기아교세포의 분화에 유의한 효능을 보였다는 것이 의미가 있다고 보여 진다.

Big data mining and validation study of microRNAs as a potential target for colon cancer prevention

강진욱 전주대학교 대학원 2016 국내석사

MicroRNA(miRNAs)는 작은 non-coding RNA 분자로써, mRNA를 불안정하게 하는 방법을 통해 표적으로 하는 mRNA를 분해하거나 번역을 억제하는 역할을 한다. miRNA는 세포사멸사, 세포증식, 세포분열, 발암 등을 포함하는 세포의 다양한 생물학적 과정들을 조절한다. 하나의 miRNA는 다양한 표적유전자를 억제할 수 있고 하나의 유전자는 다양한 miRNA에의해 조절될 수 있다. 다양한 암에서의 miRNA 발현 패턴의 변화는 세포의 발암과정과 관련이 있다. miRNA에 대한 많은 연구들은 단일 miRNA-단일 표적유전자에 대해 연구를 진행 중이다. 반면에, miRNA들의 조합이 대상으로 하는 질병에 시너지 효과를 줄 수 있는지에 대해서는 알려지지 않았다. 대장암 환자 샘플에서 공개로 제공되는 미국보건원(The Cancer Genome Atlas)으로부터 IluminaGA_miRNASeq platform을 통해 유용한 빅데이터를 추출하게 되었다. 데이터를 수집한 후, 의미 없는 데이터는 발현양 (read count/read per million)의 기준에 따라 제거되었다. 5배 이상 발현, 2.5배~5배 발현, 그 이하 등 3개의 기준에 따라 데이터는 분류되었다. 각 그룹의 miRNA들은 miRNA의 표적유전자에 관한 5개의 데이터베이스 (miRWalk, miRanda, miRDB, RNA22, and TargetScan)에 의해 분석되었다. 표적유전자와 miRNA간의 중요한 데이터는 출판된 논문들을 통해 재확인 되었다. 선별된 데이터들은 RET, Smad4, PTEN, Smad4, P53 등 암억제 유전자들과 miRNA들 간의 긍정적인 상호연관성에 관한 중요한 단서를 제공한다. 데이터 마이닝의 결과를 확인하기 위해 검증 실험을 수행한 결과, 대장암 세포주에서 miR-335-3p의 과발현과 암억제유전자인 PTEN과 연관성을 확인할 수 있었다. miR-335-3p의 억제는 PTEN의 단백질 발현을 증가시켰고, miR-335-3p의 PTEN 조절능력을 확인할 수 있었다. 데이터 마이닝 분석을 통해 도출된 miRNA들은 이전연구의 결과보다 더 효과적일 수 있다. 이는, 각각의 공통 miRNA관한 anti-sense oligonucleotide의 혼합을 통해 시너지 효과를 증명할 것이기 때문이다. miRNA에 관한 각각의 anti-sense는 새로운 기능성식품 뿐만아니라 동물의 사료로 발전할 수 있는 큰 가능성을 가지고 있다. 키워드 : 마이크로RNA, 빅데이터분석, 타겟유전자, 대장암예방

옥수수수염, 들깻잎 및 포도줄기 복합추출물의 피부면역기능 개선효과에 대한 연구

황영민 전주대학교 대학원 2017 국내석사

This study was conducted to evaluate the synergistic protective effects of mixtures of corn silk, perilla leaf and grape stem extracts (CPG mixture) in UVB-induced mouse skin damage and pruritus induced by compound 48/80. To assess this, anti-oxidant, anti-inflammation and anti-pruritus activities of CPG mixture were investigated. Results showed that CPG mixture exhibited potent 2,2′-azino-bi(3-ethylbenzthiazoline-6-sulfonicacid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activities and reducing power. In addition CPG mixture suppressed nitric oxide production in lipopolysaccharide-stimulated Raw 264.7 cells. CPG mixture exhibited the strongest suppressive effects of erythema and melanin index as well as skin damage compared to treatment with ascorbic acid (AA). Upon the examination of histological sections, CPG mixture significantly reduced epidermal thickness and the infiltration of mast cells in UVB-induced skin damaged mice. Treatment with CPG mixture also inhibited glutathione (GSH) depletion and lipid peroxidation in UVB-irradiated mice. Furthermore the treatment with CPG mixture inhibited compound 48/80-induced scratching behavior in mice. In conclusion, these results indicated that CPG mixture can be used as a functional food and cosmetic ingredient for treatment of UVB-related skin damage and compound 48/80-induced scratching. Keywords: CPG mixture, anti-oxidant, anti-inflammation, scratching, erythema, melanin.