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논문 상세정보

누드마우스에 주입된 DU-145 전립샘암에서 엑스선과 중성자선에 의한 HIF-$1\\alpha$, Bcl-2, Bax 발현의 차이

Different Expressions of HIF-$1\\alpha$, Bcl-2 and Baxin DU145 Prostate Cancer Cells Transplanted in Nude Mouse between X-Ray and Neutron Irradiation

초록

목 적: 전립샘암 세포주 DU 145에서 엑스선과 중성자선에 의한 HIF-$1\alpha$와 아포프토시스 발현의 차이를 비교함으로써 엑스선과 중성자선의 방사선생물학적 차이의 기전을 알아보고자 한다. 대상 및 방법: 누드 마우스에 DU 145 전립샘암 세포주를 주입한 후 2 Gy 엑스선, 10 Gy 엑스선, 0.6 Gy 중성자선, 3.3 Gy 중성자선을 각각 조사했다. 엑스선을 조사한 군과 중성자선을 조사한 군에서 HIF-$1\alpha$, Bcl-2, Bax, 아포프토시스 발현 정도를 면역조직화학 염색과 western blotting을 이용하여 비교하였다. 아포토시스의 정도는 terminal deoxynucleotidyl biotin-dUTP nick end labeling (TUNEL) 염색을 이용하여 비교하였다. 결 과: 방사선 조사 1일째, X선을 조사한 군과 비교했을 때, 중성자선을 조사한 군에서 HIF-$1\alpha$와 Bcl-2의 발현은 감소하였고, Bax와 아포프토시스 세포의 수는 증가하였다. Bcl-2/Bax 비는 중성자선을 조사한 군에서 의미 있게 감소하였다. 이러한 HIF-$1\alpha$, Bcl-2, Bax, Bcl-2/Bax 비, 아포프토시스 발현의 차이는 방사선 조사 5일째에도 동일하게 유지되어 나타났다. 또한, HIF-$1\alpha$ 발현은 방사선 조사 5일째 Bcl-2 (p=0.031), Bax (p=0.037), TUNEL (p=0.016) 발현과 연관성을 보였다. 결 론: 중성자선 조사한 경우 엑스선에 비해 HIF-$1\alpha$와 Bcl-2 발현, Bcl-2/Bax 비가 감소하고, Bax 발현은 증가하였다. 중성자선 치료의 광자선과 다른 방사선생물학적인 반응은 HIF-$1\alpha$와 그로 인한 아포프토시스 관련 단백질 발현의 차이와 연관성이 있을 것으로 생각한다.

Abstract

Purpose: To investigate the radiobiologic effects of neutron and X-ray irradiation on DU-145 prostate carcinoma cells by identifying the differences of HIF-$1\alpha$ expression and apoptosis. Materials and Methods: Nude mice were injected with the human prostate cancer cell line, DU-145, and then irradiated with 2 Gy and 10 Gy X-rays, or 0.6 Gy and 3.3 Gy neutrons, respectively. The mice were sacrificed at 24 hours and 120 hours after irradiation. The expression levels of HIF-$1\alpha$, Bcl-2 and Bax were compared with immunohistochemical staining and western blotting. The apoptotic indexes were compared with the Terminal deoxynucleotidyl biotin-dUTP nick and labeling (TUNEL) assay. Results: At day 1, HIF-$1\alpha$ and Bcl-2 expression decreased, while Bax expression and the number of TUNEL positive cells increased in neutron irradiated groups for the control and X-ray irradiated groups. The Bcl-2/Bax ratio was significantly lower in the neutron irradiated groups regardless of dose (p=0.001). The same pattern of the differences in the expressions of the HIF-$1\alpha$, Bcl-2, Bax, Bcl-2/Bax ratio, and apoptotic indexes were indentified at day 5. HIF-$1\alpha$ expression was related with Bcl-2 (p=0.031), Bax (p=0.037) expressions and the apoptotic indexes (p=0.016) at day 5. Conclusion: Neutron irradiation showed a decrease in HIF-$1\alpha$, Bcl-2 expression, and Bcl-2/Bax ratio, but increased Bax expression regardless of dose. This study suggests that the differences radiobiological responses between photon and neutron irradiation may be related to different HIF-$1\alpha$ expression and subsequent apoptotic protein expressions.

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