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Establishment and Characterization of Nineteen Human Colorectal Cancer Cell Lines
인체 대장암 세포주 19종의 특성 분석

DC Field Value Language
dc.contributor.advisor구자록-
dc.contributor.author김현수-
dc.date.accessioned2020-05-07T05:39:04Z-
dc.date.available2020-05-07T05:39:04Z-
dc.date.issued2020-
dc.identifier.other000000160069-
dc.identifier.urihttp://dcollection.snu.ac.kr/common/orgView/000000160069ko_KR
dc.description학위논문(석사)--서울대학교 대학원 :의과대학 협동과정 종양생물학전공,2020. 2. 구자록.-
dc.description.abstractColorectal cancer (CRC) was the second most prevalent cancer and cause of cancer-related death in Korea in 2017. Known for its difficulty in management, studies to find effective treatments for CRC are on-going. To assist further in vitro study, nineteen CRC cell lines, SNU-1566, SNU-1983, SNU-2172, SNU-2297, SNU-2303, SNU-2353B, SNU-2359, SNU-2373B, SNU-2407, SNU-2423, SNU-2431, SNU-2465, SNU-2493, SNU-2536C, SNU-2621B, SNU-NCC-61, SNU-NCC-81, SNU-NCC-376, SNU-NCC-377 derived from Korean patients were characterized. Growth rate varied between CRC cell lines where the slowest growing cell line had doubling time of 5.759 days and the fastest growing cell line had its doubling time of 1.346 days. DNA finger printing analysis was carried out for authentication of cell lines. Whole exome sequencing was performed to detect prevalent mutations. As a result, single nucleotide variation, frame shift, inframe deletions and insertions, start codon deletion and splice stop codon mutation of various genes were found and classified based on their pathogenicity reports. In addition, cell viability assay was carried out to measure the sensitivity to 24 anti-cancer drugs that are widely used for various cancers including anti-metabolite, kinase inhibitor, histone deacetylase inhibitor, alkylating inhibitor, topoisomerase inhibitor. As the results of microsatellite instability (MSI) test, 5 CRC cell lines showed MSI, of which the MLH1orMSH6 genes were mutated. Furthermore, western blot analysis showed that the expression of HER2 was increased in nine of CRC and EGFR was augmented in five of CRC cell lines. The expression of mismatch repair, MLH1 and MSH2 was significantly decreased in five of CRC cell lines. The newly established CRC cell lines can be used in investigation of the biological characteristics of CRC, particularly for investigations related to gene alterations associated with CRC.-
dc.description.abstract대장암은 한국에서 발병률과 환자 사망률이 높은 암 중 하나이다. 치료 효율이 낮아 이를 극복하고자 관련 연구들이 활발히 진행되고 있다. 대장암 관련 연구에 일조하고자 본 논문에서는 한국인 대장암 환자 유래 대장암 19개 세포주 SNU-1566, SNU-1983, SNU-2172, SNU-2297, SNU-2303, SNU-2353B, SNU-2359, SNU-2373B, SNU-2407, SNU-2423, SNU-2431, SNU-2465, SNU-2493, SNU-2536C, SNU-2621B, SNU-NCC-61, SNU-NCC-81, SNU-NCC-376, SNU-NCC-377의 특성 분석 연구를 진행하였다. 각각의 세포주마다 성장 속도가 다르게 나타났으며 가장 느리게 자라는 세포주는 두 배로 증식하기까지5.759 일이 소요되었고 가장 빠르게 자라는 세포주는 1.346 일이 소요되었다. 각 세포주가 겹치지 않고 각각 다른 세포주라는 것을 확인을 위하여 DNA finger printing을 하였습니다. Whole exome sequencing을 이용하여 각각의 세포주에 대한 돌연변이 검사를 하였다. 이 검사 결과로 얻은 돌연변이 목록은 돌연변이의 종류 (single nucleotide variation, frame shift, inframe deletions and insertions, start codon deletion, splice stop codon mutation) 또는 임상적 영향에 따라 분류하였다. 또한 19개 대장암 세포주에 24종의 항암제를 처리하여 AUC값을 통하여 항암제 민감성을 확인하였다. MSI 연구 결과 5개 대장암 세포주에서 MLH1, MSH6 유전자에서 돌연변이가 나타나고 암화 과정과 관련성이 높았다. 그리고 19개 대장암 세포주에서 HER2와 EGFR 발현이 각각 9개와 5개 세포주에서 높았으며, MLH1 와 MSH2 발현은 MSI라고 판별된 세포주들에서 발현이 되지 않았다. SNU-NCC-81은 항문 유래 흑색종으로서 임상적으로도 매우 드물며 위험한 종양 이다. 본 논문에서 확인한 대장암 세포주의 성장 특징, 돌연변이 목록, 항암제에 대한 민감성, 세포 내 단백질 발현 변화 결과가 후속 연구에 활용되어 도움이 될 것이라고 생각된다.-
dc.description.tableofcontentsIntroduction 1

Material and methods 4
Establishment of Cell Lines and Cell culture 4
Mycoplasma test 4
Growth properties and morphology in vitro 5
DNA fingerprinting 6
Drug sensitivity test 6
Western blotting analysis 7
Whole exome sequencing 8
MSI test 9

Results 10
General characteristics of CRC cell lines 10
Morphology in vitro and growth properties of CRC cell lines 13
DNA fingerprinting of 19 CRC cell lines. 16
MSI analysis of 19 CRC cell lines. 18
The expression level of growth factor receptor and EMT protein of 19 CRC cell lines 21
Anticancer drug response of 19 CRC cell lines. 23
Mutation analysis of CRC-related genes. 25

Discussion 37
References 44
Abstract in Korean 51
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dc.language.isoeng-
dc.publisher서울대학교 대학원-
dc.subject.ddc616.994-
dc.titleEstablishment and Characterization of Nineteen Human Colorectal Cancer Cell Lines-
dc.title.alternative인체 대장암 세포주 19종의 특성 분석-
dc.typeThesis-
dc.typeDissertation-
dc.contributor.department의과대학 협동과정 종양생물학전공-
dc.description.degreeMaster-
dc.date.awarded2020-02-
dc.identifier.uciI804:11032-000000160069-
dc.identifier.holdings000000000042▲000000000044▲000000160069▲-
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College of Medicine/School of Medicine (의과대학/대학원)Program in Cancer Biology (협동과정-종양생물학전공)Theses (Master's Degree_협동과정-종양생물학전공)
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