녹차 폴리페놀이 감마선조사에 의한 백혈병과 림프구모세포의 손상에 미치는 영향의 차이

Differential Effects of Green Tea Polyphenol in the ${\gamma}-irradiation$ Induced Human Leukemic and Lymphoblastic Cell Damage

  • 정환정 (원광대학교 의과대학 핵의학교실) ;
  • 김은미 (원광대학교 의과대학 핵의학교실) ;
  • 민정준 (전남대학교 의과대학 핵의학교실) ;
  • 범희승 (전남대학교 의과대학 핵의학교실) ;
  • 김영호 (조선대학교 자연대학 생물학과) ;
  • 정영도 (전남대학교 의과대학 미생물학교실) ;
  • 김창근 (원광대학교 의과대학 핵의학교실)
  • Jeong, Hwan-Jeong (Department of Nuclear Medicine, Wonkwang University School of Medicine) ;
  • Kim, Eun-Mi (Department of Nuclear Medicine, Wonkwang University School of Medicine) ;
  • Min, Jung-Jun (Department of Nuclear Medicine, Wonkwang University School of Medicine) ;
  • Bom, Hee-Seung (Department of Nuclear Medicine, Wonkwang University School of Medicine) ;
  • Kim, Young-Ho (Department of Biology, College of Natural Science, Chosun University) ;
  • Jeong, Young-Do (Department of Microbiology, Chonnam National Univerity School of Medicine) ;
  • Kim, Chang-Guhn (Department of Nuclear Medicine, Wonkwang University School of Medicine)
  • 발행 : 2003.10.30

초록

목적: 녹차 추출물(GTPP)은 암 예방과 암세포 성장억제 효과 외에 항산화제의 효능이 있는 것으로 알려져 있다. 이번 연구에서는 암세포에 감마 방사선을 이용하여 치료하는 경우 GTPP를 첨가함으로써 암세포 억제 증폭 효과와 정상세포에서의 방사선방호 효과가 함께 나타나는지 여부를 확인하고자 하였다. 대상 및 방법: GTPP (EGCG > 45%, catechin 80% 포함)를 사람 백혈병 세포주인 HL60과 사람 림프구 모세포인 NC37에 방사선을 쪼이기 전에 미리 첨가한 후 실험을 하였다. 두 세포주에서 각각의 GTPP 농도와 방사선양에 따라서 생존능을 평가하여 GTPP 농도와 방사능 양을 결정하였으며, 이를 이용하여 GTPP농도에 따른 NC37에서 방사선방호 효과와 HL60에서의 암세포 억제 효과에 대한 실험을 시행하였다. 결과: NC37과 HL60 세포주에서 암세포 억제효과를 보이면서 정상세포에 큰 영향을 미치지 않는 방사선 조사량은 1 Gy와 3 Gy정도이고, GTPP의 농도는 $10{\mu}g/ml$$20{\mu}g/ml$였다. NC37 세포주에서 GTPP를 농도별로 첨가하고 1 Gy와 3 Gy의 방사선을 각각 조사하였을 때 $10{\mu}g/ml$의 경우에는 3 Gy를 조사한 경우에만 대조군에 비해 유의한 차이를 보였으며(1 Gy;P=0.126, 3 Gy;P=0.010), $20{\mu}g/ml$를 첨가한 경우는 1 Gy와 3 Gy를 조사한 군 모두 대조군과 비교하여 유의한 차이를 보이지 않았다(1 Gy;P=0.946, 3 Gy;P=0.096). HL60 세포주에서는 방사선 조사량에 큰 상관없이 GTPP의 농도의 증가에 따라 암세포 성장이 크게 억제됨을 알 수 있었다($1\;Gy;\;10{\mu}g/ml;\;69.0{\pm}1.7%\;vs\;20{\mu}g/ml;\;42.4{\pm}1.3%,\;3\;Gy;\;10{\mu}g/ml;\;66.9{\pm}3.9%\;vs\;20{\mu}g/ml;\;44.2{\pm}1.6%$). 결론: 시험관 내 실험을 통하여 내부 방사선 치료를 시행하는 경우 GTPP를 첨가함으로써 정상세포에서 방사선방호 효과와 암세포에서 성장 억제 효과를 동시에 나타낼 수 있음을 확인할 수 있었다. 이러한 결과를 바탕으로 추후 생체 내 실험을 통한 녹차 추출물의 정상 세포에 대한 방사선방호 작용을 확인할 필요가 있을 것으로 사료된다. 녹차 추출물을 제공하여 주신 태평양 녹차 연구소의 소정 박사님께 감사드립니다.

Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.

키워드

참고문헌

  1. Otsuka T, Ogo T, Eto T, Asano Y, Suganuma M, Niho Y. Growth inhibition of leukemic cells by (-)-epigallocatechin gallate, the main constituent of green tea. Life Sci 1998;63:1397-403 https://doi.org/10.1016/S0024-3205(98)00406-8
  2. Yang GY, Liao J, Kim K, Yurkow EJ, Yang CS. Inhibition of growth and induction of apoptosis in human cancer cell lines by tea polyphenols. Carcinogenesis 1998;19:611-6 https://doi.org/10.1093/carcin/19.4.611
  3. Chen ZP, Schell JB, Ho CT, Chen KY. Green tea epigallocatechin gallate shows a pronounced growth inhibitory effect on cancerous cells but not on their normal counterparts. Cancer Lett 1998;129:173-9 https://doi.org/10.1016/S0304-3835(98)00108-6
  4. Suganuma M, Okabe S, Kai Y, Sueoka N, Sueoka E, Fujiki H. Synergistic effects of (-)-epigallocatechin gallate with (-)-epicatechin, sulindac, or tamoxifen on cancer-preventive activity in the human lung cancer cell line PC-9. Cancer Res 1999;59:44-7
  5. Bohuslavizki KH, Brenner W, Klutmann S, Hubner RH, Lassmann S, Feyerabend B, et al. Radioprotection of salivary glands by amifostine in high-dose radioiodine therapy. J Nucl Med 1998;39:1237-42
  6. Steele VE, Kelloff GJ, Balentine D, Boone CW, Mehta R, Bagheri D, et al. Comparative chemopreventive mechanisms of green tea, black tea and selected polyphenol extracts measured by in vitro bioassays. Carcinogenesis 2000 ;21:63-7 https://doi.org/10.1093/carcin/21.1.63
  7. Parshad R, Sanford KK, Price FM, Steele VE, Tarone RE, Kelloff GJ, et al. Protective action of plant polyphenols on radiation-induced chromatid breaks in cultured human cells. Anticancer Res 1998;18:3263-6
  8. Castillo J, Benavente-Garcia O, Lorente J, Alcaraz M, Redondo A, Ortuno A, et al. Antioxidant activity and radioprotective effects against chromosomal damage induced in vivo by X-rays of flavan-3-ols (Procyanidins) from grape seeds (Vitis vinifera): comparative study versus other phenolic and organic compounds. J Agric Food Chem 2000;48:1738-45 https://doi.org/10.1021/jf990665o
  9. Benua RS, Cicale NR, Sonenberg M, Rawson RW. The relation of radioiodine dosimetry to results and complications in the treatment of metastatic thyroid cancer. AJR. 1962;87:171-82
  10. Kim JC, Yoon JH, Bom HS, Jaegal YJ, Song HC, Min JJ, et al. Development and assessment individual maximum permissible dose method of I-131 therapy in high risk patients with differentiated papillary thyroid cancer. Kor J Nucl Med 2003;37:110-9
  11. Maxon HR, Thomas SR, Hertzberg VS, Kereiakes JG, Chen IW, Sperling MI, et al. Relation between effective radiation dose and outcome of radioiodine therapy for thyroid cancer. N Engl J Med 1983;309:937-41 https://doi.org/10.1056/NEJM198310203091601
  12. Karam M, Gianoukakis A, Feustel PJ, Cheema A, Postal ES, Cooper JA. Influence of diagnostic and therapeutic doses on thyroid remnant ablation rates. Nucl Med Commun 2003;24:489-95 https://doi.org/10.1097/00006231-200305000-00002
  13. Uchida S, Ozaki M, Suzuki K, Shikita M. Radioprotective effects of (-)-epigallocatechin 3-O-gallate (green-tea tannin) in mice. Life Sci 1992;50:147-52 https://doi.org/10.1016/0024-3205(92)90296-2
  14. Kim S, Lee MJ, Hong J, Li C, Smith TJ, Yang GY, et al. Plasma and tissue levels of tea catechins in rats and mice during chronic consumption of green tea polyphenols. Nutr Cancer 2000;37:41-8 https://doi.org/10.1207/S15327914NC3701_5