자살유전자가 도입된 인간신경줄기세포를 이용한 뇌종양치료법
Human neural stem cells target and deliver therapeutic gene to glioblastoma
- 주제(키워드) neural stem cell , glioma , human , migration , suicide gene
- 주제(KDC) 513
- 주제(DDC) 616
- 발행기관 아주대학교 일반대학원
- 지도교수 김승업
- 발행년도 2007
- 학위수여년월 2007. 2
- 학위명 석사
- 학과 및 전공 일반대학원 신경과학기술과정
- 본문언어 영어
초록/요약
신경줄기세포는 질병과 상처에 의해 손상된 신경세포를 대체할 수 있고 이를 위해 종양을 포함한 각각의 손상된 부분으로 이동할 수 있는 능력을 가지고 있다. 한편 자살유전자인 cytosine deaminase (CD) 유전자는 독성이 없는 전구약물인 5-Fluorocytosine을 5-Fluorouracil로 변환시킨 후 세포자살을 유도한다고 알려져 있다. 그래서 본 연구에서는 신경줄기세포를 이용한 종양치료를 위해 v-myc 유전자로 불멸화 시킨 F3 신경줄기세포주에 자살유전자인 cytosine deaminase(CD) 유전자를 도입하여 F3.CD 세포주를 만들었다. 그리고 동물종양모델의 종양 외 지역에 이식한 후 5-FC를 투여한 결과 F3.CD 세포주는 종양지역으로 이동하여 5-FU로 변환시킨 후 스스로 자살함으로써 주변의 뇌종양 세포까지 함께 사멸시켰고 이로 인해 대조군에 비해 종양의 크기가 90% 까지 줄어드는 효과를 볼 수 있었다. 이러한 결과는 종양치료에 있어서 뇌종양 뿐만 아니라 전이된 여러 종양에 적용할 수 있다는 점을 시사하고 있다.
more초록/요약
Neural stem cells (NSCs) could serve as a replacement source for cells in the central nervous system damaged by disease or injury. In addition to their potential value as a source of replacement cells, NSCs possess a remarkable ability to home sites of pathology, including tumors, in vivo. Selective migration of NSCs has been demonstrated in brain tumor models when administered intracranially or intravenously, and localization to brain tumor loci. Because of this ability to migrate to tumor sites, engineered NSCs could be used to deliver chemotherapeutic agents. In the previous studies, therapeutic efficacy has been demonstrated in animal models of glioma using murine NSCs to deliver the gene encoding E. coli cytosine deaminase (CD) to convert systemically administered 5-fluorocytosine (5-FC) to the chemotherapeutic agent 5-fluorouracil (5-FU). We investigated whether human NSCs might also be useful as a tumor-selective delivery vector for treatment of metastatic tumor. Previously we have generated a stable cell line of human NSC using a retroviral vector encoding v-myc oncogene (HB1.F3). F3 human NSC line was transduced to carry gene encoding CD. In animal models of glioblastoma in the brain, we show that F3. CD human NSCs target intracranial tumors and after systemic treatment with 5-FC, there were dramatic reductions in tumor masses. This NSC-enzyme-prodrug strategy increased median survival of animals markedly compared to controls, suggesting that NSC-mediated anti-tumor therapy may have therapeutic applications for treatment of patients with metastatic cancer in the brain.
more목차
Ⅰ. INTRODUCTION = 1
Ⅱ. MATERAIL AND METHODS = 4
Cell culture = 4
CD recombinant plasmids and F3 cell lines infection with pMSCV-puro/CD = 4
RNA extraction and RT-PCR = 5
Sensitivity of cells to 5FC and 5FU = 6
In vitro ‘Bystander effect’ experiments = 7
In vitro migration assay = 7
Confirmation of chemoattractant ligands and receptors = 8
In vivo migration assay = 9
In vivo intracerebral implantation of cancer cell lines = 10
Ⅲ. RESULTS = 13
Generation and Characterization of CD-transduced HB1.F3 cell lines = 13
Effects of the 5-Fluorocytosine on CD-transduced HB1.F3 CD cell lines = 13
Effects of the 5-Fluorouracil on tumor cell, CD- and CD+ HB1.F3 cell lines = 14
Bystander effect of tumor cell with F3.CD = 14
Migratory capacity of hNSCs in vitro = 15
Confirmation of chemoattractant ligand and receptors = 15
NSCs migrate throughout and “Surround” Tumor in vivo = 16
In vivo therapeutic efficacy of HB1.F3CD = 16
Ⅳ. DISCUSSION = 18
Ⅴ. REFERENCES = 36
국문 요약 = 42
