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 Irradiation원문보기
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.
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.
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, ...
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.
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.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
제안 방법
In the report using the intermediate grade non-Hodgkin’s lymphoma cell line, WSU-DLCL2, which is resistant to chemotherapy, the authors demonstrated that neutron radiation at doses of 0.5 and 1.5 Gy caused apoptosis as a result of down-regulation of Bcl-2 and increased expression of Bax at 24, 48, 72 hours compared with the control.
From the pooled data on the radiation sensitivity of the neutron beam of the cyclotron,9,10) the biologically equivalent doses between X-ray and neutron were determined. The animals were divided into five groups (n=10 in each group): the tumor-treated non-irradiated group as a control, the tumor- treated 2 Gy, the tumor-treated 10 Gy X-ray irradiated group, the tumor-treated 0.6 Gy neutron (biologically equivalent dose to 2 Gy X-ray), the tumor treated 3.3 Gy neutron (biologically equivalent dose to 10 Gy X-ray) irradiated group.
The animals were housed under specific pathogen-free conditions (20±2℃) lighting from 07:00 to 19:00 and were supplied with auto-claved food and water ad libitum.
The sections were rinsed and visualized using Converter-POD with 0.02% 3,3’-diaminoben-zidine (DAB).
대상 데이터
The human prostate carcinoma cell line DU-145 was purchased from the Korean Cell Line Bank (Seoul, Korea). The cells were cultured in Roswell Park Memorial Institute 1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (Gibco BRL) at 37℃ in 5% CO2, 95% O2 in a humidified cell incubator.
성능/효과
For the comparison between groups, one-way analysis of variance between groups (ANOVA) and Duncan’s post-hoc test were performed and differences among groups were considered statistically signi-ficant at p＜0.05.
In conclusion, the results that the expression of HIF-1α after neutron treatment was significantly suppressed compared with X-ray may suggest neutron irradiation for prostate cancer patients may have radiobiological benefits through suppression of apoptotic protein expression.
This results suggest that HIF-1α and apoptosis are associated in DU-145 prostate cancer cells and there was significant difference of apoptosis between neutron and X-ray.
참고문헌 (26)
Halperin EC. Particle therapy and treatment of cancer. Lancet Oncol 2006;7:676-685
Wambersie A, Hendry J, Gueulette J, Gahbauer R, Potter R, Gregoire V. Radiobiological rationale and patient selection for high-LET radiation in cancer therapy. Radiother Oncol 2004;73 Suppl 2:S1-14
Forman JD, Yudelev M, Bolton S, Tekyi-Mensah S, Maughan R. Fast neutron irradiation for prostate cancer. Cancer Metastasis Rev 2002;21:131-135
Russell KJ, Caplan RJ, Laramore GE, et al. Photon versus fast neutron external beam radiotherapy in the treatment of locally advanced prostate cancer: results of a randomized prospective trial. Int J Radiat Oncol Biol Phys 1994;28:47-54
Hall EJ, Giaccia AJ. Radiobiology for the radiologist. 6th ed. Philadelphia: Lippincott Williams & Wilkins, 2006:109
Zhong H, Semenza GL, Simons JW, De Marzo AM. Up-regulation of hypoxia-inducible factor 1alpha is an early event in prostate carcinogenesis. Cancer Detect Prev 2004;28:88-93
Strowbridge A, Wall N, Aref A, et al. Neutron radiation induces the modulation of BCL-2 and BAX protein resulting in apoptosis in a chemotherapy resistant intermediate grade non-Hodgkin's lymphoma cell line. Proc Am Soc Clin Oncol 2000;19:abstr 2601
Eom KY, Wu HG, Park HJ, et al. Evaluation of biological characteristics of neutron beam generated from MC50 cyclotron. J Korean Soc Ther Radiol Oncol 2006;24:280-284
Lee HJ, Kim JS, Moon C, Kim JC, Jo SK, Kim SH. Relative biological effectiveness of fast neutrons in a multiorgan assay for apoptosis in mouse. Environ Toxicol 2008;23:233-239
Moeller BJ, Richardson RA, Dewhirst MW. Hypoxia and radiotherapy: opportunities for improved outcomes in cancer treatment. Cancer Metastasis Rev 2007;26:241-248
Moeller BJ, Cao Y, Li CY, Dewhirst MW. Radiation activates HIF-1 to regulate vascular radiosensitivity in tumors:role of reoxygenation, free radicals, and stress granules. Cancer Cell 2004;5:429-441
Holl V, Coelho D, Weltin D, Dufour P, Gueulette J, Bischoff P. Ex vivo determination of the effect of whole-body exposure to fast neutrons on murine spleen cell viability and apoptosis. Radiat Res 2000;154:301-306
Vral A, Cornelissen M, Thierens H, et al. Apoptosis induced by fast neutrons versus 60Co gamma-rays in human peripheral blood lymphocytes. Int J Radiat Biol 1998;73:289-295
Meijer AE, Kronqvist US, Lewensohn R, Harms-Ringdahl M. RBE for the induction of apoptosis in human peripheral lymphocytes exposed in vitro to high-LET radiation generated by accelerated nitrogen ions. Int J Radiat Biol 1998;73:169-177
Kyprianou N, King ED, Bradbury D, Rhee JG. Bcl-2 over-expression delays radiation-induced apoptosis without affecting the clonogenic survival of human prostate cancer cells. Int J Cancer 1997;70:341-348
Mackey TJ, Borkowski A, Amin P, Jacobs SC, Kyprianou N. bcl-2/bax ratio as a predictive marker for therapeutic response to radiotherapy in patients with prostate cancer. Urology 1998;52:1085-1090
Khor LY, Moughan J, Al-Saleem T, et al. Bcl-2 and Bax expression predict prostate cancer outcome in men treated with androgen deprivation and radiotherapy on radiation therapy oncology group protocol 92-02. Clin Cancer Res 2007;13:3585-3590
Khor LY, Desilvio M, Li R, et al. Bcl-2 and bax expression and prostate cancer outcome in men treated with radiotherapy in Radiation Therapy Oncology Group protocol 86-10. Int J Radiat Oncol Biol Phys 2006;66:25-30
※ AI-Helper는 부적절한 답변을 할 수 있습니다.