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Mechanism of Synergy between RAS and HOX in Leukemia Induction

Title
Mechanism of Synergy between RAS and HOX in Leukemia Induction
Other Titles
백혈병 유발에 있어 RAS와 HOX의 상승효과 및 그 작용기전에 관한 연구
Author
이소현
Alternative Author(s)
Lee, So Hyun
Advisor(s)
정희용
Issue Date
2015-02
Publisher
한양대학교
Degree
Master
Abstract
RAS protein is an important signaling protein which relays information from environment to the cell. However, RAS mutant that are constitutively active are found to be involved in the incidence of various cancers. Previous study has shown that constitutive activation of RAS alone is inadequate for promoting tumor malignancy. Therefore, the effort of finding RAS partner-in-crime in promoting malignant tumor will elucidate the mechanism of cancer advancement. In this study, we use leukemia as a model for studying complex signaling pathway between RAS and HOX genes. We hypothesize that RAS and HOX work together to induce leukemia. We co-expressed human RAS (NRASG12D or KRASQ61H) and human HOX cDNA using retroviral vector into normal mouse bone marrow cells, which were then transplanted into gamma-irradiated mice. Mice which developed leukemia were sacrificed and their genomic DNAs were isolated for genetic analysis. The cDNA inserts that contributed to the formation of leukemia were sequenced for identification. We established that HOXA1, HOXA6, HOXB6, HOXB7, HOXB9, and HOXD8 were crucial, together with NRASG12D or KRASQ61H, for inducing leukemia in vivo. Furthermore, we confirmed this finding in vitro by using colony formation assay, proliferation assay, and FACS analysis on the mouse primary BM cell with exogenous expression of RAS and HOX. This study will shed light on the molecular crosstalk between RAS and HOX genes during the development of leukemia and other malignant cancer, and may lead to the new avenue of drug target discovery for leukemia treatment.|RAS 단백질은 외부환경으로부터 세포로 정보를 전달하는 중요한 신호전달 단백질이며, 항상 활성화된 RAS 돌연변이는 다양한 암의 발생과 관련이 있다고 알려져 있다. 하지만 지금까지의 연구에서는 항상 활성화된 RAS 단독으로는 악성 종양을 촉진하는데 불충분하다는 것으로 나타났다. 그러므로, 악성 종양을 유도하는 RAS의 파트너를 찾아 암으로 발전하는 기전을 밝히기 위하여 본 연구를 수행 하였다. 본 연구실에서 기존에 수립되어진 암유전자 라이브러리 시스템을 분석하여, RAS와 HOX 유전자 사이에 복잡한 신호 전달 경로를 연구하기 위한 모델로서 백혈병을 사용했고, RAS와 HOX가 함께 작용하여 백혈병을 유발한다는 가설을 세웠다. 인간 RAS (NRASG12D 또는 KRASQ61H) 와 HOX의 cDNA를 레트로바이러스 벡터를 사용하여 정상 쥐 골수세포에서 동시 발현하도록 한 후 이 세포들을 감마를 조사한 쥐에 이식하였다. 백혈병이 발병한 쥐들의 유전체 DNA를 유전 분석을 위해 분리하였다. 백혈병 형성의 원인이 된 삽입 cDNA를 확인하기 위해 시퀀싱을 하여, HOXA1, HOXA6, HOXB6, HOXB7, HOXB9, HOXD8이 NRASG12D나 KRASQ61H와 함께 생체 내에서 백혈병을 유발하는데 중요하다는 것을 밝혔다. 뿐만 아니라, 우리는 이 결과를 RAS와 HOX를 외래 발현하는 쥐의 골수 세포를 가지고 생체 외에서 콜로니 형성 분석, 증식 분석, FACS 분석을 함으로써 이중으로 검증하였다. 그러므로, 이 연구는 백혈병과 다른 악성 종양이 발생하는 동안 RAS와 HOX 유전자 간의 분자적 크로스 토크가 있음을 밝혔으며, 추가 연구를 통하여 백혈병 치료를 위한 의약품 표적 발견의 새로운 방안이 되기를 기대한다.; RAS protein is an important signaling protein which relays information from environment to the cell. However, RAS mutant that are constitutively active are found to be involved in the incidence of various cancers. Previous study has shown that constitutive activation of RAS alone is inadequate for promoting tumor malignancy. Therefore, the effort of finding RAS partner-in-crime in promoting malignant tumor will elucidate the mechanism of cancer advancement. In this study, we use leukemia as a model for studying complex signaling pathway between RAS and HOX genes. We hypothesize that RAS and HOX work together to induce leukemia. We co-expressed human RAS (NRASG12D or KRASQ61H) and human HOX cDNA using retroviral vector into normal mouse bone marrow cells, which were then transplanted into gamma-irradiated mice. Mice which developed leukemia were sacrificed and their genomic DNAs were isolated for genetic analysis. The cDNA inserts that contributed to the formation of leukemia were sequenced for identification. We established that HOXA1, HOXA6, HOXB6, HOXB7, HOXB9, and HOXD8 were crucial, together with NRASG12D or KRASQ61H, for inducing leukemia in vivo. Furthermore, we confirmed this finding in vitro by using colony formation assay, proliferation assay, and FACS analysis on the mouse primary BM cell with exogenous expression of RAS and HOX. This study will shed light on the molecular crosstalk between RAS and HOX genes during the development of leukemia and other malignant cancer, and may lead to the new avenue of drug target discovery for leukemia treatment.
URI
https://repository.hanyang.ac.kr/handle/20.500.11754/128804http://hanyang.dcollection.net/common/orgView/200000425819
Appears in Collections:
GRADUATE SCHOOL OF BIOMEDICAL SCIENCE AND ENGINEERING[S](의생명공학전문대학원) > BIOMEDICAL SCIENCE(의생명과학과) > Theses (Master)
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