유방암 분자표지자 ECRG4가 세포생장에 미치는 영향

다국어 초록 (Multilingual Abstract)

Breast cancer is the second most common type of cancer after lung cancer, and the fifth most common cause of cancer death. The expression of multiple genes in a cancer sample may provide useful information about cancer or normal tissue. These genes are called biomarkers. In our previous study, we searched biomarkers that increased or decreased in breast cancer from the currently available gene-expression data sets. And these biomarkers were verified by relative quantitative realtime PCR. ECRG4 is one of the biomarkers and may work as a tumor suppressor gene and affect on nuclear factor κB (NF-κB) pathway. The aim of this study was to investigate whether a breast cancer biomarker influences on cell proliferation. In this study, effects of ECRG4 on cell growth is evaluated by indirectly detecting and directly counting cell numbers in cancer cell lines transiently expressing ECRG4. Before investigating function of ECRG4 in cell, we needed to select cell lines to maximize effect of ECRG4. MIApaca2, Hs578T, and MDA-MB-231 cell lines were selected by relative quantitative real time PCR and cell line screening. According to expectation, transiently transfected MIApaca2 and Hs578T cell lines with ECRG4 grow more slowly than transiently transfected cell lines with empty vectors, but MDA-MB-231 cell line expressing ECRG4 showed faster growth. Therefore, ECRG4 has opposite effect in MDA-MB-231 cell line. This phenomenon is able to be explained by different effect of Akt pathway in MDA-MB-231. In MDA-MB-231 cell, introduced Akt, a well-known oncogene, inhibits cell proliferation and plays an opposite role. Therefore, we suggest that ECRG4 protein is involved in Akt signal transduction network.

목차 (Table of Contents)

• 제1장. 서 론 1
• 제2장. 재료 및 방법 5
• 1. 세포주, E. coli 균주 및 플라스미드 5
• 2. 배지 및 배양 조건 5
• 3. 유방암세포주에서 유방암 분자 표지자의 발현 정도 비교 9
• 4. 형질 전환과 DNA 조작 12
• 5. 도입 유전자 발현의 확인 15
• 6. ECRG4 단백질의 정제 18
• 7. ECRG4 형질 전환 세포주에서 세포 생장 관찰 18
• 제3장. 결과 및 고찰 20
• 1. 유방암세포주에서 유방암 분자 표지자 발현 정도 비교 20
• 2. 완성한 벡터의 발현 확인 20
• 3. 재조합 ECRG4의 정제 21
• 4. 세포 내 발현 ECRG4의 정제 22
• 5. 역전사 PCR을 이용한 유방암세포주에서의 ECRG4 발현 확인 23
• 6. 암세포주의 세포 생장 속도 분석 23
• 제4장. 결 론 39
• 제5장. 참고문헌 42