大蒜素诱导LM-8细胞凋亡及相关基因表达的实验研究
摘要
背景和目的:近年来随着新辅助化疗在临床治疗中的广泛应用,骨肉瘤患者的总体生存率有了很大提高,但仍有部分患者对化疗药物不敏感或耐药,同时大剂量化疗药物容易导致一系列的毒副作用,寻找新的安全有效的化疗药物成为提高化疗效果的途径之一。大蒜素(Allicin)又名大蒜新素,化学名二烯丙基三硫化合物(diallyl trisulfide),是从大蒜球茎中分离出的硫化物,在中国作为药物治疗各种疾病已经有三千多年历史。许多流行病学调查发现居民食用大蒜的量与癌症发病率和死亡率呈显著负相关体外实验研究证实,大蒜素可以通过调节机体免疫功能、细胞周期、凋亡和成血管作用等抑制胃癌等多种上皮来源恶性肿瘤生长,但尚未有关于间叶组织来源如骨肉瘤细胞的报道。本论文就大蒜素对C3H小鼠骨肉瘤细胞株LM-8细胞增殖的影响及与Bcl-2、Bax凋亡蛋白表达的关系进行研究,并初步探索其诱导凋亡的机制。
方法:用水溶性四唑盐WST-8比色法检测大蒜素对LM-8细胞株生长的影响;倒置显微镜观察细胞形态变化;流式细胞仪检测细胞调亡率的变化;免疫细胞组化技术检测Bcl-2和Bax蛋白表达。
结果:大蒜素能明显抑制LM-8细胞的生长,与大蒜素的浓度和时间有关,在72h其药物半数抑制率浓度是11.09μg/ml;5μg/ml、10μg/ml和15μg/ml浓度的大蒜素作用72小时后可诱导LM-8细胞凋亡(P<0.05);大蒜素可以降低Bcl-2表达,增强Bax表达,经15μg/ml大蒜素作用72h,Bcl-2和Bax灰度值及bcl-2/bax分别为(6.61±1.07)%、(35.72±5.16)%和(0.19±0.03),对照组分别为(23.28±6.17)%、(12.52±2.18)%和1.86±0.48,与对照组比较,差异有显著性意义(P<0.05)。
结论:大蒜素可以诱导LM-8细胞凋亡,并且诱导作用呈剂量依赖性,其机制涉及到Bcl-2表达上调及Bax表达下调,有望成为临床防治骨肉瘤的理想药物。
Background and Objective:Although neoadjuvant chemotherapy has improved survival in osteosarcoma patients greatly since it was introdued in late 1970s, drug resistance and side effect caused by high-dose severely limited the therapeutic effectiveness. It is now in a high demand to find new feasible and well tolerated regimens to improve the current clinical outcomes. Allicin is a natural compound derived from the bulbodium of garlic and has been used as a remedy for a variety of diseases by Chinese tradition herbalist for thousands of years. Many epidemiological studies support the protective role of allicin against the development of certain human cancers. The anticarcinogenic and antitumorigenic characteristics appear to arise through both dose- and temporal-related changes in a number of cellular events involved with the cancer process, including those involving drug metabolism, immunocompetence, cell cycle regulation, apoptosis, and angiogenesis. The ability of allicin to block tumors from epithelium tissue such as colon, lung, breast, and liver has been proved except for osteosarcoma from mesenchymal tissue. This essay is to investigate the effects of Allicin on LM-8 cells and probe the possible mechanism.
Method:Water-soluble tetrazolium salt (WST-8) assay was used to detect the growth inhibitory effect in 24, 48 and 72h. Light microscopy was performed to observe the morphologic changes of cells. Flow cytometry was used to evaluate apoptosis. The protein expression of Bcl-2 and Bax was examined by immunohistochemistry.
Result: Allicin inhibited the growth of LM-8, and the inhibitory rate of cells treated by Allicin (11.09μg/ml) for 72h was 50%. Cells cultured with Allicin were observed to present typical apoptosis changes. Flow cytometry revealed a dose-dependent relationship. The positive protein expression rates of Bcl-2 and Bax were (7.24±1.18)% and (38.56±7.43)% respectively with Allicin of 15μg/ml at 72h, while the controls being (24.40±9.96)% and (11.28±1.31)%(p<0.05), and the value of Bcl-2/ Bax was related with the dose of Allicin(p<0.05).
Conclusion:Allicin can induce apoptosis in LM-8 cells by the downregulation of bcl-2 expression and upregulation of bax expression.
方法:用水溶性四唑盐WST-8比色法检测大蒜素对LM-8细胞株生长的影响;倒置显微镜观察细胞形态变化;流式细胞仪检测细胞调亡率的变化;免疫细胞组化技术检测Bcl-2和Bax蛋白表达。
结果:大蒜素能明显抑制LM-8细胞的生长,与大蒜素的浓度和时间有关,在72h其药物半数抑制率浓度是11.09μg/ml;5μg/ml、10μg/ml和15μg/ml浓度的大蒜素作用72小时后可诱导LM-8细胞凋亡(P<0.05);大蒜素可以降低Bcl-2表达,增强Bax表达,经15μg/ml大蒜素作用72h,Bcl-2和Bax灰度值及bcl-2/bax分别为(6.61±1.07)%、(35.72±5.16)%和(0.19±0.03),对照组分别为(23.28±6.17)%、(12.52±2.18)%和1.86±0.48,与对照组比较,差异有显著性意义(P<0.05)。
结论:大蒜素可以诱导LM-8细胞凋亡,并且诱导作用呈剂量依赖性,其机制涉及到Bcl-2表达上调及Bax表达下调,有望成为临床防治骨肉瘤的理想药物。
Background and Objective:Although neoadjuvant chemotherapy has improved survival in osteosarcoma patients greatly since it was introdued in late 1970s, drug resistance and side effect caused by high-dose severely limited the therapeutic effectiveness. It is now in a high demand to find new feasible and well tolerated regimens to improve the current clinical outcomes. Allicin is a natural compound derived from the bulbodium of garlic and has been used as a remedy for a variety of diseases by Chinese tradition herbalist for thousands of years. Many epidemiological studies support the protective role of allicin against the development of certain human cancers. The anticarcinogenic and antitumorigenic characteristics appear to arise through both dose- and temporal-related changes in a number of cellular events involved with the cancer process, including those involving drug metabolism, immunocompetence, cell cycle regulation, apoptosis, and angiogenesis. The ability of allicin to block tumors from epithelium tissue such as colon, lung, breast, and liver has been proved except for osteosarcoma from mesenchymal tissue. This essay is to investigate the effects of Allicin on LM-8 cells and probe the possible mechanism.
Method:Water-soluble tetrazolium salt (WST-8) assay was used to detect the growth inhibitory effect in 24, 48 and 72h. Light microscopy was performed to observe the morphologic changes of cells. Flow cytometry was used to evaluate apoptosis. The protein expression of Bcl-2 and Bax was examined by immunohistochemistry.
Result: Allicin inhibited the growth of LM-8, and the inhibitory rate of cells treated by Allicin (11.09μg/ml) for 72h was 50%. Cells cultured with Allicin were observed to present typical apoptosis changes. Flow cytometry revealed a dose-dependent relationship. The positive protein expression rates of Bcl-2 and Bax were (7.24±1.18)% and (38.56±7.43)% respectively with Allicin of 15μg/ml at 72h, while the controls being (24.40±9.96)% and (11.28±1.31)%(p<0.05), and the value of Bcl-2/ Bax was related with the dose of Allicin(p<0.05).
Conclusion:Allicin can induce apoptosis in LM-8 cells by the downregulation of bcl-2 expression and upregulation of bax expression.
引文
1. Wittig JC, Bickels J, Priebat D, Jelinek J, Kellar-Graney K, Shmookler B, Malawer MM. Osteosarcoma: a multidisciplinary approach to diagnosis and treatment. Am Fam Physician. 2002 Mar 15;65(6):1123-32.
2. Bacci G, Lari S. Adjuvant and neoadjuvant chemotherapy in osteosarcoma. Chir Organi Mov. 2001 Oct-Dec;86(4):253-68.
3. Knowles LM, Milner JA. Diallyl disulfide inhibits p34(cdc2) kinase activity through changes in complex formation and phosphorylation. Carcinogenesis. 2000;21:1129–34.
4. Nakagawa H, Tsuta K, Kiuchi K, Senzaki H, Tanaka K, Hioki K, Tsubura A. Growth inhibitory effects of diallyl disulfide on human breast cancer cell lines. Carcinogenesis 2001;22:891–7.
5. Xiao D, Pinto JT, Soh JW, Deguchi A, Gundersen GG, Palazzo AF, Yoon JT, Shirin H, Weinstein IB. Induction of apoptosis by the garlic-derived compound S-allylmercaptocysteine (SAMC) is associated with microtubule depolymerization and c-Jun NH2-terminal kinase 1 activation. Cancer Res 2003;63:6825–37.
6. Filomeni G, Aquilano K, Rotilio G, Ciriolo MR. Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell death induced by diallyl disulfide. Cancer Res 2003;63:5940–9.
7. Herman-Antosiewicz A, Singh SV. Signal transduction pathways leading to cell cycle arrest and apoptosis induction in cancer cells by Allium vegetable-derived organosulfur compounds: a review. Mutat Res 2004;555:121–31.
8. Xiao D, Choi S, Johnson DE, Vogel VG, Johnson CS, Trump DL, Lee YJ, Singh SV. Diallyl trisulfide-induced apoptosis in human prostate cancer cells involves c-Jun N-terminal kinase and extracellular-signal regulated kinase-mediated phosphorylation of Bcl-2. Oncogene 2004;23:5594–606.
9. Hosono T, Fukao T, Ogihara J, Ito Y, Shiba H, Seki T, Ariga T. Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of ?-tubulin. J Biol Chem 2005;280:41487–93.
10. Takezaki T, Gao CM, Wu JZ, Ding JH, Liu YT, Zhang Y, Li SP, Su P, Liu TK, TajimaK. Dieatary protective and risk factors for esophageal and stomach cancers in a low-epidemic area for stomach cancer in Jiangsu province, China: comparison with those in a high-epidemic area. Jpn J Cancer Res, 2001,92:1157-1165.
11. Fenwick G. R., Hanley A. B. The genus Allium—Part 3. Crit. Rev. Food Sci. Nutr. 1985; 23:1-73.
12. Milner J. A. Garlic: its anticarcinogenic and antitumorigenic properties. Nutr. Rev. 1996; 54:S82-S86.
13. Milner J. A. Functional foods and health promotion. J. Nutr. 1999;129(suppl.): 1395S -1397S.
14. Orekhov A. N., Grunwald J. Effects of garlic on atherosclerosis. Nutrition 1997; 13:656-663.
15. Yoshida H., Katsuzaki H., Ohta R., Ishikawa K., Fukuda H., Fujino T., Suzuki A. An organosulfur compound isolated from oil-macerated garlic extract, and its antimicrobial effect. Biosci. Biotechnol. Biochem. 1999;63:588-590.
16. Amagase H., Milner J. A. Impact of various sources of garlic and their constituents on 7,12-dimethylbenz(a)anthracene binding to mammary cell DNA. Carcinogenesis 1993; 14:1627-1631.
17. Hussain S. P., Jannu L. N., Rao A. R. Chemopreventive action of garlic on methylcholanthrene-induced carcinogenesis in the uterine cervix of mice. Cancer Lett 1990;49:175-180.
18. Ip C., Lisk D. J., Stoewsand G. S. Mammary cancer prevention by regular garlic and selenium-enriched garlic. Nutr. Cancer 1992;7:279-286.
19. Liu J. Z., Lin R. I., Milner J. A. Inhibition of 7,12-dimethylbenz(a)-anthracene induced mammary tumors and DNA adducts by garlic powder. Carcinogenesis 1992;13:1847-1851.
20. Shukla Y., Singh A., Srivastava B. Inhibition of carcinogen-induced activity of gamma-glutamyl transpeptidase by certain dietary constituents in mouse skin. Biomed. Environ. Sci. 1999;12:110-115.
21. Song K., Milner J. A. Heating garlic inhibits its ability to suppress 7, 12-dimethylbenz(a) anthracene-induced DNA adduct formation in rat mammary tissue.J. Nutr. 1999;129: 657-661.
22. Sumiyoshi H., Wargovich M. J. Chemoprevention of 1,2-dimethylhydrazine-induced colon cancer in mice by natural occurring organosulfur compounds. Cancer Res 1990;50: 5084-5087.
23. Wargovich M. J., Woods C., Eng V. W., Stephens L. C., Gray K. Chemoprevention of N-nitrosomethylbenzylamine-induced esophageal cancer in rats by the naturally occurring thioether, diallyl sulfide. Cancer Res 1988;48:6872-6875.
24. Siegers CP, Steffen B, R?bke A, Pentz R. The effects of garlic preparations against human tumor cell proliferation. Phytomedicine. 1999 Mar;6(1):7-11.
25. Frantz DJ, Hughes BG, Nelson DR, Murray BK, Christensen MJ. Cell cycle arrest and differential gene expression in HT-29 cells exposed to an aqueous garlic extract. Nutr Cancer. 2000;38(2):255-64.
26. Filomeni G, Aquilano K, Rotilio G, Ciriolo MR. Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell death induced by diallyl disulfide. Cancer Res. 2003 Sep 15;63(18):5940-9.
27. Knowles LM, Milner JA. Diallyl disulfide induces ERK phosphorylation and alters gene expression profiles in human colon tumor cells. J Nutr. 2003 Sep;133(9):2901-6.
28. Wu CC, Chung JG, Tsai SJ, Yang JH, Sheen LY. Differential effects of allyl sulfides from garlic essential oil on cell cycle regulation in human liver tumor cells. Food Chem Toxicol. 2004 Dec;42(12):1937-47.
29.王美岭,韩金祥.大蒜与肿瘤.中国肿瘤1994;3:6-8
30. Sakamoto K, Lawson LD, Milner JA. Allyl sulfides from garlic suppress the in vitro proliferation of human A549 lung tumor cells. Nutr Cancer. 1997;29(2):152-6.
31. Bours V, Bentires-Alj M, Hellin AC, Viatour P, Robe P, Delhalle S, et al. Nuclear factor-κB, cancer, and apoptosis. Biochem Pharmacol 2000; 60:1085-1089.
32.扈启宽,李文梅,赵喜荣,吕有勇.大蒜素对胃癌细胞NFκB活性的影响[J].解放军药学学报,2002, 18(1):7～10.
33. Wong WW, MacDonald S, Langler RF, Penn LZ. Novel synthetic organosulfur compounds induce apoptosis of human leukemic cells. Anticancer Res2000;20:1367-74.
34. Sigounas G, Hooker JL, Li W, Anagnostou A, Steiner M. Sigounas G, Hooker JL, Li W, Anagnostou A, Steiner M. Nutr Cancer. 1997;28(2):153-9.
35. Sengupta A, Ghosh S, Bhattacharjee S, Das S. Indian food ingredients and cancer prevention - an experimental evaluation of anticarcinogenic effects of garlic in rat colon. Asian Pac J Cancer Prev. 2004 Apr-Jun;5(2):126-32.
36.孙丽,王绪,潘友金.大蒜素对人胃腺癌SGC-7901细胞株端粒酶活性和细胞凋亡的影响[J].解放军医学杂志,2003,28(5):445-6.
37.李晓光,谢锦玉,李文梅,季加孚,赵敏,崔建涛,孙梅,吕有勇.大蒜油对人胃癌细胞恶性增生能力和细胞间通讯的影响.中国科学(C辑) 1998;28:366.
38. Lee JM, Bernstein A. Lee JM, Bernstein A. p53 mutations increase resistance to ionizing radiation. p53 mutations increase resistance to ionizing radiation.
39.李晓光,谢锦玉,李文梅,季加孚,赵敏,崔建涛,孙梅,吕有勇.大蒜油对人胃癌细胞恶性增生能力和细胞间通讯的影响.中国科学(C辑) 1998;28:366.
40. Kwon KB, Yoo SJ, Ryu DG, Yang JY, Rho HW, Kim JS, Park JW, Kim HR, Park BH. Induction of apoptosis by diallyl disulfide through activation of caspase-3 in human leukemia HL-60 cells. Biochem Pharmacol. 2002 Jan 1;63(1):41-7.
41. Green D R,Reed J C.Mitochondrion and apoptosis[J].Science,1998,281(28):1309-12.
42. Yang E, Zha J, Jockel J, Boise LH, Thompson CB, Korsmeyer SJ. Yang E, Zha J, Jockel J, Boise LH, Thompson CB, Korsmeyer SJ. Cell. 1995 Jan 27;80(2):285-91.
43. Kaseta MK, Khaldi L, Gomatos IP, Tzagarakis GP, Alevizos L, Leandros E, Papagelopoulos PJ, Soucacos PN. Prognostic value of bax, bcl-2, and p53 staining in primary osteosarcoma. J Surg Oncol. 2008 Mar 1;97(3):259-66.
1. Rydholm A. Prognostic factors in soft tissue sarcoma. Acta Orthop Scand Suppl. 1997 Feb;273:148-55.
2. Rosen G. Preoperative (neoadjuvant) chemotherapy for osteogenic sarcoma: a ten year experience.Orthopedics. 1985 May;8(5):659-64.
3. Fagioli F, Aglietta M, Tienghi A, Ferrari S, Brach del Prever A, Vassallo E, Palmero A, Biasin E, Bacci G, Picci P, Madon E. High-dose chemotherapy in the treatment of relapsed osteosarcoma: an Italian sarcoma group study.J Clin Oncol. 2002 Apr 15;20(8):2150-6
4. Bacci G, Ferrari S, Bertoni F, Picci P, Bacchini P, Longhi A, Donati D, Forni C, Campanacci L, Campanacci M. Histologic response of high-grade nonmetastatic osteosarcoma of the extremity to chemotherapy. Clin Orthop Relat Res. 2001 May;(386): 186-96.
5. Reddick WE ,Wang S ,Xiong X , et al . Dynamic magnetic resonance imaging of regional contrast access as an additional prognostic factor in pediatric osteosarcoma [J]. Cancer ,2001 ,91(12) :2230 - 7.
6. Wittig JC, Bickels J, Priebat D, Jelinek J, Kellar-Graney K, Shmookler B, Malawer MM. Osteosarcoma: a multidisciplinary approach to diagnosis and treatment. Am Fam Physician. 2002 Mar 15;65(6):1123-32.
7. Rosen G, Nirenberg A. Neoadjuvant chemotherapy for osteogenic sarcoma: a five year follow-up (T-10) and preliminary report of new studies (T-12).Prog Clin Biol Res. 1985;201:39-51.
8. Rosen G. Neoadjuvant chemotherapy for osteogenic sarcoma: a model for the treatment of other highly malignant neoplasms.Recent Results Cancer Res. 1986;103:148-57.
9. Eilber FR, Rosen G. Adjuvant chemotherapy for osteosarcoma.Semin Oncol. 1989 Aug;16(4):312-22.
10. Bielack S, Kempf-Bielack B, Schwenzer D, Birkfellner T, Delling G, Ewerbeck V, Exner GU, Fuchs N, G?bel U, Graf N, Heise U, Helmke K, von Hochstetter AR, Jürgens H, Maas R, Münchow N, Salzer-Kuntschik M, Treuner J, Veltmann U, WernerM, Winkelmann W, Zoubek A, Kotz R. [Neoadjuvant therapy for localized osteosarcoma of extremities. Results from the Cooperative osteosarcoma study group COSS of 925 patients] Klin Padiatr. 1999 Jul-Aug;211(4):260-70.
11. Bacci G, Ferrari S, Bertoni F, Ruggieri P, Picci P, Longhi A, Casadei R, Fabbri N, Forni C, Versari M, Campanacci M. Long-term outcome for patients with nonmetastatic osteosarcoma of the extremity treated at the istituto ortopedico rizzoli according to the istituto ortopedico rizzoli/osteosarcoma-2 protocol: an updated report.J Clin Oncol. 2000 Dec 15;18(24):4016-27.
12. Trapeznikov NN, Aliev MD, Machak GN, Kushlinskii NE, Sokolovskii VA, Tepliakov VV, Kharatishvili TK, Sobolevskii VA, Musaev ER, Mistakopulo NF. [Treatment of limb osteosarcoma at the turn of the century (half century of experience in research)] Vestn Ross Akad Med Nauk. 2001;(9):46-9.
13. Bacci G, Longhi A, Versari M, Mercuri M, Briccoli A, Picci P. Prognostic factors for osteosarcoma of the extremity treated with neoadjuvant chemotherapy: 15-year experience in 789 patients treated at a single institution.Cancer. 2006 Mar 1;106(5):1154-61.
14. de Wilt JH, Bout A, Eggermont AM, van Tiel ST, de Vries MW, ten Hagen TL, de Roos WK, Valerio D, van der Kaaden ME. Adenovirus-mediated interleukin 3 beta gene transfer by isolated limb perfusion inhibits growth of limb sarcoma in rats.Hum Gene Ther. 2001 Mar 20;12(5):489-502.
15. de Wilt JH, Bout A, Eggermont AM, van Tiel ST, de Vries MW, ten Hagen TL, de Roos WK, Valerio D, van der Kaaden ME. Adenovirus-mediated interleukin 3 beta gene transfer by isolated limb perfusion inhibits growth of limb sarcoma in rats.Hum Gene Ther. 2001 Mar 20;12(5):489-502.
16. Lejeune FJ, Liénard D, Matter M, Rüegg C. Efficiency of recombinant human TNF in human cancer therapy.Cancer Immun. 2006 Mar 22;6:6.
17. van Etten B, de Vries MR, van IJken MG, Lans TE, Guetens G, Ambagtsheer G, van Tiel ST, de Boeck G, de Bruijn EA, Eggermont AM, ten Hagen TL. Degree of tumour vascularity correlates with drug accumulation and tumour response upon TNF-alpha-based isolated hepatic perfusion.Br J Cancer. 2003 Jan 27;88(2):314-9.
18. Lejeune FJ, Pujol N, Liénard D, Mosimann F, Raffoul W, Genton A, Guillou L, Landry M, Chassot PG, Chiolero R, Bischof-Delaloye A, Leyvraz S, Mirimanoff RO, Bejkos D, Leyvraz PF. Limb salvage by neoadjuvant isolated perfusion with TNFalpha and melphalan for non-resectable soft tissue sarcoma of the extremities.Eur J Surg Oncol. 2000 Nov;26(7):669-78.
19. Lejeune FJ, Liénard D, Matter M, Rüegg C. Efficiency of recombinant human TNF in human cancer therapy. Cancer Immun. 2006 Mar 22;6:6.
20.范镜林,杨劲松,李鼎锋,崔秋,袁海涛,黄宗贵.皮下埋植式动脉介入联合咖啡因治疗恶性骨肿瘤14例分析.中国肿瘤临床与康复. 2005年03期.
21. Arpaci F, Ataergin S, Ozet A, Erler K, Basbozkurt M, Ozcan A, Komurcu S, Ozturk B, Celasun B, Kilic S, Kuzhan O. The feasibility of neoadjuvant high-dose chemotherapy and autologous peripheral blood stem cell transplantation in patients with nonmetastatic high grade localized osteosarcoma: results of a phase II study. Cancer. 2005 Sep 1;104 (5):1058-65.
22. Sauerbrey A, Bielack S, Kempf-Bielack B, Zoubek A, Paulussen M, Zintl F. High-dose chemotherapy (HDC) and autologous hematopoietic stem cell transplantation (ASCT) as salvage therapy for relapsed osteosarcoma. Bone Marrow Transplant. 2001 May; 27 (9):933-7.
23. Bacci G, Loro L, Longhi A, Bertoni F, Bacchini P, Versari M, Picci P, Serra M. No correlation between methotrexate serum level and histologic response in the pre-operative treatment of extremity osteosarcoma. Anticancer Drugs. 2006 Apr;17(4):411-5.
24. Tomoda R, Seto M, Hioki Y, Sonoda J, Matsumine A, Kusuzaki K, Uchida A. Low-dose methotrexate inhibits lung metastasis and lengthens survival in rat osteosarcoma. Clin Exp Metastasis. 2005;22(7):559-64. Epub 2006 Feb 11.
25. Chu F, Chou PM, Zheng X, Mirkin BL, Rebbaa A. Control of multidrug resistance gene mdr1 and cancer resistance to chemotherapy by the longevity gene sirt1. Cancer Res. 2005 Nov 15;65(22):10183-7.
26. Hattinger CM, Reverter-Branchat G, Remondini D, Castellani GC, Benini S, Pasello M, Manara MC, Scotlandi K, Picci P, Serra M. Genomic imbalances associated withmethotrexate resistance in human osteosarcoma cell lines detected by comparative genomic hybridization-based techniques. Eur J Cell Biol. 2003 Sep;82(9):483-93.
27. Bruheim S, Bruland OS, Breistol K, Maelandsmo GM, Fodstad O. Human osteosarcoma xenografts and their sensitivity to chemotherapy. Pathol Oncol Res. 2004;10(3):133-41. Epub 2004 Sep 25.
28. Martín-Duque P, Quintanilla M, McNeish I, Lopes R, Romero J, Romero D, Lemoine NR, Ramón y Cajal S, Vassaux G.. Caspase-1 as a radio- and chemo-sensitiser in vitro and in vivo. Int J Mol Med. 2006 May;17(5):841-7.
29. Pasello M, Hattinger CM, Stoico G, Manara MC, Benini S, Geroni C, Mercuri M, Scotlandi K, Picci P, Serra M. 4-Demethoxy-3'-deamino-3'-aziridinyl-4' -methylsulphonyl- daunorubicin (PNU-159548): a promising new candidate for chemotherapeutic treatment of osteosarcoma patients. Eur J Cancer. 2005 Sep;41(14): 2184-95.
30. Pool BL, Berger M, Schlehofer JR, Wingen F. In vivo and in vitro investigations on biological effects of aromatic bis-(2-chloroethyl)amino-bisphosphonic acids, new agents proposed for chemotherapy of bone tumors: cytostatic activity in rat osteosarcoma; toxicity and genotoxicity in liver and bone marrow; mutagenicity in S. typhimurium. Invest New Drugs. 1988 Jun;6(2):67-78.
31. Pool BL, Berger M, Schlehofer JR, Wingen F. Antitumor effect of nutrient synergy on human osteosarcoma cells U-2OS, MNNG-HOS and Ewing's sarcoma SK-ES.1. Oncol Rep. 2005 Feb;13(2):253-7.
32. Kubista B, Trieb K, Sevelda F, Toma C, Arrich F, Heffeter P, Elbling L, Sutterlüty H, Scotlandi K, Kotz R, Micksche M, Berger W. Anticancer effects of zoledronic acid against human osteosarcoma cells. J Orthop Res. 2006 Jun;24(6):1145-52.
33. Kubista B, Trieb K, Sevelda F, Toma C, Arrich F, Heffeter P, Elbling L, Sutterlüty H, Scotlandi K, Kotz R, Micksche M, Berger W. RhoA GTPase inactivation by statins induces osteosarcoma cell apoptosis by inhibiting p42/p44-MAPKs-Bcl-2 signaling independently of BMP-2 and cell differentiation. Cell Death Differ. 2006 Feb 10.
34. Alleva R, Benassi MS, Pazzaglia L, Tomasetti M, Gellert N, Borghi B, Neuzil J, Picci P. Alpha-tocopheryl succinate alters cell cycle distribution sensitising humanosteosarcoma cells to methotrexate-induced apoptosis. Cancer Lett. 2006 Feb 8;232(2):226-35.
35. Kang HJ, Jo C, Kwon JH, Son JH, An BJ, Byun MW. Antioxidant and cancer cell proliferation inhibition effect of citrus pectin-oligosaccharide prepared by irradiation. J Med Food. 2006 Fall;9(3):313-20.
36. Zou DM, Brewer M, Garcia F, Feugang JM, Wang J, Zang R, Liu H, Zou C. Cactus pear: a natural product in cancer chemoprevention. Nutr J. 2005 Sep 8;4:25.
37. Aires V, Adote S, Hichami A, Moutairou K, Boustani ES, Khan NA. Modulation of intracellular calcium concentrations and T cell activation by prickly pear polyphenols. Mol Cell Biochem. 2004 May;260(1-2):103-10.
38. Su CC, Lin JG, Li TM, Chung JG, Yang JS, Ip SW, Lin WC, Chen GW. Curcumin-induced apoptosis of human colon cancer colo 205 cells through the. production of ROS, Ca2+ and the activation of caspase-3. Anticancer Res. 2006 Nov-Dec;26(6B):4379-89.
39. Holmes FA, Madden T, Newman RA, Valero V, Theriault RL, Fraschini G, Walters RS, Booser DJ, Buzdar AU, Willey J, Hortobagyi GN. Sequence-dependent alteration of doxorubicin pharmacokinetics by paclitaxel in a phase I study of paclitaxel and doxorubicin in patients with metastatic breast cancer. J Clin Oncol. 1996 Oct;14(10): 2713-21.
40.王兵,王国俊,徐钧.氧化苦参碱对肿瘤诱导血管内皮细胞增殖的抑制作用[J].实用肿瘤杂志,2000,15(5):297-300.
41.周钢,杨述华,何宇.大蒜素诱导LM-8细胞凋亡及相关基因表达的实验研究.中国中医骨伤科杂志. 2007年08期.
42.杨迪生,解先宽,叶招明,陶惠民.腺病毒介导反义c-myc增强骨肉瘤细胞对顺铂化疗敏感性的实验研究.中华外科杂志. 2005 Jun 15;43(12):799-802.
43.高大新,张海燕,吕刚,李永军,黄涛,王岩峰. Survivin反义寡核苷酸联合顺铂治疗裸鼠荷骨肉瘤.中华实验外科杂志. 2005 Nov;22(8): 1003-4.
44. Van Valen F, Fulda S, Sch?fer KL, Truckenbrod B, Hotfilder M, Poremba C, Debatin KM, Winkelmann W. Selective and nonselective toxicity of TRAIL/Apo2L combinedwith chemotherapy in human bone tumour cells vs. normal human cells. Int J Cancer. 2003 Dec 20;107(6):929-40.
45. Yang DS, Miao XD, Ye ZM, Xu YS. Synergistic induction of apoptosis by the combination of TRAIL and low dose adriamycin in human osteosarcoma cell line U2OS. Zhonghua Zhong Liu Za Zhi. 2005 Oct;27(10):595-7.
46. Nardin A, Lefebvre ML, Labroquère K, Faure O, Abastado JP. Liposomal muramyl tripeptide phosphatidylethanolamine: Targeting and activating macrophages for adjuvant treatment of osteosarcoma. Curr Cancer Drug Targets. 2006 Mar;6(2):123-33.
47. Sherr CJ. Principles of tumor suppression. Cell. 2004 Jan 23;116(2):235-46.
48. McCormick F. Cancer gene therapy: Fringe or cutting edge? Nat Rev Cancer,2001, 1:130-41.
49. Van Meir EG, Polverini PJ, Chazin VR, Su Huang HJ, de Tribolet N, Cavenee WK. Release of an inhibitor of angiogenesis upon induction of wild type p53 expression in glioblastoma cells. Nat Genet. 1994 Oct;8(2):171-6.
50. Takaoka A, Hayakawa S, Yanai H, Stoiber D, Negishi H, Kikuchi H, Sasaki S, Imai K, Shibue T, Honda K, Taniguchi T. Integration of interferon-alpha/beta signalling to p53 responses in tumour suppression and antiviral defence. Nature 2003;424:516–523.
51. Tataria M, Quarto N, Longaker MT, Sylvester KG. Absence of the p53 tumor suppressor gene promotes osteogenesis in mesenchymal stem cells. J Pediatr Surg. 2006 Apr;41(4): 624-32; discussion 624-32.
52. Graat HC, Wuisman PI, van Beusechem VW, Carette JE, Gerritsen WR, Bras J, Schaap GR, Kaspers GJ, Ogose A, Gu W, Kawashima H, Hotta T. Coxsackievirus and adenovirus receptor expression on primary osteosarcoma specimens and implications for gene therapy with recombinant adenoviruses. Clin Cancer Res. 2005 Mar 15;11(6):2445-7; author reply 2447-8.
53. Ternovoi VV, Curiel DT, Smith BF, Siegal GP. Adenovirus-mediated p53 tumor suppressor gene therapy of osteosarcoma. Lab Invest. 2006 Aug;86(8):748-66. Epub 2006 Jun 5. Review.
54. Kwon KB, Yoo SJ, Ryu DG, Yang JY, Rho HW, Kim JS, Park JW, Kim HR, Park BH. Induction of apoptosis by diallyl disulfide through activation of caspase-3 in humanleukemia HL-60 cells. Biochem Pharmacol. 2002 Jan 1;63(1):41-7.
55. Green R, Reed C. Mitochondria and apoptosis. Science, 1998;281(6);1309-1312
56. Kaseta MK, Khaldi L, Gomatos IP, Tzagarakis GP, Alevizos L, Leandros E, Papagelopoulos PJ, Soucacos PN. Kaseta MK, Khaldi L, Gomatos IP, Tzagarakis GP, Alevizos L, Leandros E, Papagelopoulos PJ, Soucacos PN. J Surg Oncol. 2008 Mar 1;97(3):259-66.
2. Bacci G, Lari S. Adjuvant and neoadjuvant chemotherapy in osteosarcoma. Chir Organi Mov. 2001 Oct-Dec;86(4):253-68.
3. Knowles LM, Milner JA. Diallyl disulfide inhibits p34(cdc2) kinase activity through changes in complex formation and phosphorylation. Carcinogenesis. 2000;21:1129–34.
4. Nakagawa H, Tsuta K, Kiuchi K, Senzaki H, Tanaka K, Hioki K, Tsubura A. Growth inhibitory effects of diallyl disulfide on human breast cancer cell lines. Carcinogenesis 2001;22:891–7.
5. Xiao D, Pinto JT, Soh JW, Deguchi A, Gundersen GG, Palazzo AF, Yoon JT, Shirin H, Weinstein IB. Induction of apoptosis by the garlic-derived compound S-allylmercaptocysteine (SAMC) is associated with microtubule depolymerization and c-Jun NH2-terminal kinase 1 activation. Cancer Res 2003;63:6825–37.
6. Filomeni G, Aquilano K, Rotilio G, Ciriolo MR. Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell death induced by diallyl disulfide. Cancer Res 2003;63:5940–9.
7. Herman-Antosiewicz A, Singh SV. Signal transduction pathways leading to cell cycle arrest and apoptosis induction in cancer cells by Allium vegetable-derived organosulfur compounds: a review. Mutat Res 2004;555:121–31.
8. Xiao D, Choi S, Johnson DE, Vogel VG, Johnson CS, Trump DL, Lee YJ, Singh SV. Diallyl trisulfide-induced apoptosis in human prostate cancer cells involves c-Jun N-terminal kinase and extracellular-signal regulated kinase-mediated phosphorylation of Bcl-2. Oncogene 2004;23:5594–606.
9. Hosono T, Fukao T, Ogihara J, Ito Y, Shiba H, Seki T, Ariga T. Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of ?-tubulin. J Biol Chem 2005;280:41487–93.
10. Takezaki T, Gao CM, Wu JZ, Ding JH, Liu YT, Zhang Y, Li SP, Su P, Liu TK, TajimaK. Dieatary protective and risk factors for esophageal and stomach cancers in a low-epidemic area for stomach cancer in Jiangsu province, China: comparison with those in a high-epidemic area. Jpn J Cancer Res, 2001,92:1157-1165.
11. Fenwick G. R., Hanley A. B. The genus Allium—Part 3. Crit. Rev. Food Sci. Nutr. 1985; 23:1-73.
12. Milner J. A. Garlic: its anticarcinogenic and antitumorigenic properties. Nutr. Rev. 1996; 54:S82-S86.
13. Milner J. A. Functional foods and health promotion. J. Nutr. 1999;129(suppl.): 1395S -1397S.
14. Orekhov A. N., Grunwald J. Effects of garlic on atherosclerosis. Nutrition 1997; 13:656-663.
15. Yoshida H., Katsuzaki H., Ohta R., Ishikawa K., Fukuda H., Fujino T., Suzuki A. An organosulfur compound isolated from oil-macerated garlic extract, and its antimicrobial effect. Biosci. Biotechnol. Biochem. 1999;63:588-590.
16. Amagase H., Milner J. A. Impact of various sources of garlic and their constituents on 7,12-dimethylbenz(a)anthracene binding to mammary cell DNA. Carcinogenesis 1993; 14:1627-1631.
17. Hussain S. P., Jannu L. N., Rao A. R. Chemopreventive action of garlic on methylcholanthrene-induced carcinogenesis in the uterine cervix of mice. Cancer Lett 1990;49:175-180.
18. Ip C., Lisk D. J., Stoewsand G. S. Mammary cancer prevention by regular garlic and selenium-enriched garlic. Nutr. Cancer 1992;7:279-286.
19. Liu J. Z., Lin R. I., Milner J. A. Inhibition of 7,12-dimethylbenz(a)-anthracene induced mammary tumors and DNA adducts by garlic powder. Carcinogenesis 1992;13:1847-1851.
20. Shukla Y., Singh A., Srivastava B. Inhibition of carcinogen-induced activity of gamma-glutamyl transpeptidase by certain dietary constituents in mouse skin. Biomed. Environ. Sci. 1999;12:110-115.
21. Song K., Milner J. A. Heating garlic inhibits its ability to suppress 7, 12-dimethylbenz(a) anthracene-induced DNA adduct formation in rat mammary tissue.J. Nutr. 1999;129: 657-661.
22. Sumiyoshi H., Wargovich M. J. Chemoprevention of 1,2-dimethylhydrazine-induced colon cancer in mice by natural occurring organosulfur compounds. Cancer Res 1990;50: 5084-5087.
23. Wargovich M. J., Woods C., Eng V. W., Stephens L. C., Gray K. Chemoprevention of N-nitrosomethylbenzylamine-induced esophageal cancer in rats by the naturally occurring thioether, diallyl sulfide. Cancer Res 1988;48:6872-6875.
24. Siegers CP, Steffen B, R?bke A, Pentz R. The effects of garlic preparations against human tumor cell proliferation. Phytomedicine. 1999 Mar;6(1):7-11.
25. Frantz DJ, Hughes BG, Nelson DR, Murray BK, Christensen MJ. Cell cycle arrest and differential gene expression in HT-29 cells exposed to an aqueous garlic extract. Nutr Cancer. 2000;38(2):255-64.
26. Filomeni G, Aquilano K, Rotilio G, Ciriolo MR. Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell death induced by diallyl disulfide. Cancer Res. 2003 Sep 15;63(18):5940-9.
27. Knowles LM, Milner JA. Diallyl disulfide induces ERK phosphorylation and alters gene expression profiles in human colon tumor cells. J Nutr. 2003 Sep;133(9):2901-6.
28. Wu CC, Chung JG, Tsai SJ, Yang JH, Sheen LY. Differential effects of allyl sulfides from garlic essential oil on cell cycle regulation in human liver tumor cells. Food Chem Toxicol. 2004 Dec;42(12):1937-47.
29.王美岭,韩金祥.大蒜与肿瘤.中国肿瘤1994;3:6-8
30. Sakamoto K, Lawson LD, Milner JA. Allyl sulfides from garlic suppress the in vitro proliferation of human A549 lung tumor cells. Nutr Cancer. 1997;29(2):152-6.
31. Bours V, Bentires-Alj M, Hellin AC, Viatour P, Robe P, Delhalle S, et al. Nuclear factor-κB, cancer, and apoptosis. Biochem Pharmacol 2000; 60:1085-1089.
32.扈启宽,李文梅,赵喜荣,吕有勇.大蒜素对胃癌细胞NFκB活性的影响[J].解放军药学学报,2002, 18(1):7～10.
33. Wong WW, MacDonald S, Langler RF, Penn LZ. Novel synthetic organosulfur compounds induce apoptosis of human leukemic cells. Anticancer Res2000;20:1367-74.
34. Sigounas G, Hooker JL, Li W, Anagnostou A, Steiner M. Sigounas G, Hooker JL, Li W, Anagnostou A, Steiner M. Nutr Cancer. 1997;28(2):153-9.
35. Sengupta A, Ghosh S, Bhattacharjee S, Das S. Indian food ingredients and cancer prevention - an experimental evaluation of anticarcinogenic effects of garlic in rat colon. Asian Pac J Cancer Prev. 2004 Apr-Jun;5(2):126-32.
36.孙丽,王绪,潘友金.大蒜素对人胃腺癌SGC-7901细胞株端粒酶活性和细胞凋亡的影响[J].解放军医学杂志,2003,28(5):445-6.
37.李晓光,谢锦玉,李文梅,季加孚,赵敏,崔建涛,孙梅,吕有勇.大蒜油对人胃癌细胞恶性增生能力和细胞间通讯的影响.中国科学(C辑) 1998;28:366.
38. Lee JM, Bernstein A. Lee JM, Bernstein A. p53 mutations increase resistance to ionizing radiation. p53 mutations increase resistance to ionizing radiation.
39.李晓光,谢锦玉,李文梅,季加孚,赵敏,崔建涛,孙梅,吕有勇.大蒜油对人胃癌细胞恶性增生能力和细胞间通讯的影响.中国科学(C辑) 1998;28:366.
40. Kwon KB, Yoo SJ, Ryu DG, Yang JY, Rho HW, Kim JS, Park JW, Kim HR, Park BH. Induction of apoptosis by diallyl disulfide through activation of caspase-3 in human leukemia HL-60 cells. Biochem Pharmacol. 2002 Jan 1;63(1):41-7.
41. Green D R,Reed J C.Mitochondrion and apoptosis[J].Science,1998,281(28):1309-12.
42. Yang E, Zha J, Jockel J, Boise LH, Thompson CB, Korsmeyer SJ. Yang E, Zha J, Jockel J, Boise LH, Thompson CB, Korsmeyer SJ. Cell. 1995 Jan 27;80(2):285-91.
43. Kaseta MK, Khaldi L, Gomatos IP, Tzagarakis GP, Alevizos L, Leandros E, Papagelopoulos PJ, Soucacos PN. Prognostic value of bax, bcl-2, and p53 staining in primary osteosarcoma. J Surg Oncol. 2008 Mar 1;97(3):259-66.
1. Rydholm A. Prognostic factors in soft tissue sarcoma. Acta Orthop Scand Suppl. 1997 Feb;273:148-55.
2. Rosen G. Preoperative (neoadjuvant) chemotherapy for osteogenic sarcoma: a ten year experience.Orthopedics. 1985 May;8(5):659-64.
3. Fagioli F, Aglietta M, Tienghi A, Ferrari S, Brach del Prever A, Vassallo E, Palmero A, Biasin E, Bacci G, Picci P, Madon E. High-dose chemotherapy in the treatment of relapsed osteosarcoma: an Italian sarcoma group study.J Clin Oncol. 2002 Apr 15;20(8):2150-6
4. Bacci G, Ferrari S, Bertoni F, Picci P, Bacchini P, Longhi A, Donati D, Forni C, Campanacci L, Campanacci M. Histologic response of high-grade nonmetastatic osteosarcoma of the extremity to chemotherapy. Clin Orthop Relat Res. 2001 May;(386): 186-96.
5. Reddick WE ,Wang S ,Xiong X , et al . Dynamic magnetic resonance imaging of regional contrast access as an additional prognostic factor in pediatric osteosarcoma [J]. Cancer ,2001 ,91(12) :2230 - 7.
6. Wittig JC, Bickels J, Priebat D, Jelinek J, Kellar-Graney K, Shmookler B, Malawer MM. Osteosarcoma: a multidisciplinary approach to diagnosis and treatment. Am Fam Physician. 2002 Mar 15;65(6):1123-32.
7. Rosen G, Nirenberg A. Neoadjuvant chemotherapy for osteogenic sarcoma: a five year follow-up (T-10) and preliminary report of new studies (T-12).Prog Clin Biol Res. 1985;201:39-51.
8. Rosen G. Neoadjuvant chemotherapy for osteogenic sarcoma: a model for the treatment of other highly malignant neoplasms.Recent Results Cancer Res. 1986;103:148-57.
9. Eilber FR, Rosen G. Adjuvant chemotherapy for osteosarcoma.Semin Oncol. 1989 Aug;16(4):312-22.
10. Bielack S, Kempf-Bielack B, Schwenzer D, Birkfellner T, Delling G, Ewerbeck V, Exner GU, Fuchs N, G?bel U, Graf N, Heise U, Helmke K, von Hochstetter AR, Jürgens H, Maas R, Münchow N, Salzer-Kuntschik M, Treuner J, Veltmann U, WernerM, Winkelmann W, Zoubek A, Kotz R. [Neoadjuvant therapy for localized osteosarcoma of extremities. Results from the Cooperative osteosarcoma study group COSS of 925 patients] Klin Padiatr. 1999 Jul-Aug;211(4):260-70.
11. Bacci G, Ferrari S, Bertoni F, Ruggieri P, Picci P, Longhi A, Casadei R, Fabbri N, Forni C, Versari M, Campanacci M. Long-term outcome for patients with nonmetastatic osteosarcoma of the extremity treated at the istituto ortopedico rizzoli according to the istituto ortopedico rizzoli/osteosarcoma-2 protocol: an updated report.J Clin Oncol. 2000 Dec 15;18(24):4016-27.
12. Trapeznikov NN, Aliev MD, Machak GN, Kushlinskii NE, Sokolovskii VA, Tepliakov VV, Kharatishvili TK, Sobolevskii VA, Musaev ER, Mistakopulo NF. [Treatment of limb osteosarcoma at the turn of the century (half century of experience in research)] Vestn Ross Akad Med Nauk. 2001;(9):46-9.
13. Bacci G, Longhi A, Versari M, Mercuri M, Briccoli A, Picci P. Prognostic factors for osteosarcoma of the extremity treated with neoadjuvant chemotherapy: 15-year experience in 789 patients treated at a single institution.Cancer. 2006 Mar 1;106(5):1154-61.
14. de Wilt JH, Bout A, Eggermont AM, van Tiel ST, de Vries MW, ten Hagen TL, de Roos WK, Valerio D, van der Kaaden ME. Adenovirus-mediated interleukin 3 beta gene transfer by isolated limb perfusion inhibits growth of limb sarcoma in rats.Hum Gene Ther. 2001 Mar 20;12(5):489-502.
15. de Wilt JH, Bout A, Eggermont AM, van Tiel ST, de Vries MW, ten Hagen TL, de Roos WK, Valerio D, van der Kaaden ME. Adenovirus-mediated interleukin 3 beta gene transfer by isolated limb perfusion inhibits growth of limb sarcoma in rats.Hum Gene Ther. 2001 Mar 20;12(5):489-502.
16. Lejeune FJ, Liénard D, Matter M, Rüegg C. Efficiency of recombinant human TNF in human cancer therapy.Cancer Immun. 2006 Mar 22;6:6.
17. van Etten B, de Vries MR, van IJken MG, Lans TE, Guetens G, Ambagtsheer G, van Tiel ST, de Boeck G, de Bruijn EA, Eggermont AM, ten Hagen TL. Degree of tumour vascularity correlates with drug accumulation and tumour response upon TNF-alpha-based isolated hepatic perfusion.Br J Cancer. 2003 Jan 27;88(2):314-9.
18. Lejeune FJ, Pujol N, Liénard D, Mosimann F, Raffoul W, Genton A, Guillou L, Landry M, Chassot PG, Chiolero R, Bischof-Delaloye A, Leyvraz S, Mirimanoff RO, Bejkos D, Leyvraz PF. Limb salvage by neoadjuvant isolated perfusion with TNFalpha and melphalan for non-resectable soft tissue sarcoma of the extremities.Eur J Surg Oncol. 2000 Nov;26(7):669-78.
19. Lejeune FJ, Liénard D, Matter M, Rüegg C. Efficiency of recombinant human TNF in human cancer therapy. Cancer Immun. 2006 Mar 22;6:6.
20.范镜林,杨劲松,李鼎锋,崔秋,袁海涛,黄宗贵.皮下埋植式动脉介入联合咖啡因治疗恶性骨肿瘤14例分析.中国肿瘤临床与康复. 2005年03期.
21. Arpaci F, Ataergin S, Ozet A, Erler K, Basbozkurt M, Ozcan A, Komurcu S, Ozturk B, Celasun B, Kilic S, Kuzhan O. The feasibility of neoadjuvant high-dose chemotherapy and autologous peripheral blood stem cell transplantation in patients with nonmetastatic high grade localized osteosarcoma: results of a phase II study. Cancer. 2005 Sep 1;104 (5):1058-65.
22. Sauerbrey A, Bielack S, Kempf-Bielack B, Zoubek A, Paulussen M, Zintl F. High-dose chemotherapy (HDC) and autologous hematopoietic stem cell transplantation (ASCT) as salvage therapy for relapsed osteosarcoma. Bone Marrow Transplant. 2001 May; 27 (9):933-7.
23. Bacci G, Loro L, Longhi A, Bertoni F, Bacchini P, Versari M, Picci P, Serra M. No correlation between methotrexate serum level and histologic response in the pre-operative treatment of extremity osteosarcoma. Anticancer Drugs. 2006 Apr;17(4):411-5.
24. Tomoda R, Seto M, Hioki Y, Sonoda J, Matsumine A, Kusuzaki K, Uchida A. Low-dose methotrexate inhibits lung metastasis and lengthens survival in rat osteosarcoma. Clin Exp Metastasis. 2005;22(7):559-64. Epub 2006 Feb 11.
25. Chu F, Chou PM, Zheng X, Mirkin BL, Rebbaa A. Control of multidrug resistance gene mdr1 and cancer resistance to chemotherapy by the longevity gene sirt1. Cancer Res. 2005 Nov 15;65(22):10183-7.
26. Hattinger CM, Reverter-Branchat G, Remondini D, Castellani GC, Benini S, Pasello M, Manara MC, Scotlandi K, Picci P, Serra M. Genomic imbalances associated withmethotrexate resistance in human osteosarcoma cell lines detected by comparative genomic hybridization-based techniques. Eur J Cell Biol. 2003 Sep;82(9):483-93.
27. Bruheim S, Bruland OS, Breistol K, Maelandsmo GM, Fodstad O. Human osteosarcoma xenografts and their sensitivity to chemotherapy. Pathol Oncol Res. 2004;10(3):133-41. Epub 2004 Sep 25.
28. Martín-Duque P, Quintanilla M, McNeish I, Lopes R, Romero J, Romero D, Lemoine NR, Ramón y Cajal S, Vassaux G.. Caspase-1 as a radio- and chemo-sensitiser in vitro and in vivo. Int J Mol Med. 2006 May;17(5):841-7.
29. Pasello M, Hattinger CM, Stoico G, Manara MC, Benini S, Geroni C, Mercuri M, Scotlandi K, Picci P, Serra M. 4-Demethoxy-3'-deamino-3'-aziridinyl-4' -methylsulphonyl- daunorubicin (PNU-159548): a promising new candidate for chemotherapeutic treatment of osteosarcoma patients. Eur J Cancer. 2005 Sep;41(14): 2184-95.
30. Pool BL, Berger M, Schlehofer JR, Wingen F. In vivo and in vitro investigations on biological effects of aromatic bis-(2-chloroethyl)amino-bisphosphonic acids, new agents proposed for chemotherapy of bone tumors: cytostatic activity in rat osteosarcoma; toxicity and genotoxicity in liver and bone marrow; mutagenicity in S. typhimurium. Invest New Drugs. 1988 Jun;6(2):67-78.
31. Pool BL, Berger M, Schlehofer JR, Wingen F. Antitumor effect of nutrient synergy on human osteosarcoma cells U-2OS, MNNG-HOS and Ewing's sarcoma SK-ES.1. Oncol Rep. 2005 Feb;13(2):253-7.
32. Kubista B, Trieb K, Sevelda F, Toma C, Arrich F, Heffeter P, Elbling L, Sutterlüty H, Scotlandi K, Kotz R, Micksche M, Berger W. Anticancer effects of zoledronic acid against human osteosarcoma cells. J Orthop Res. 2006 Jun;24(6):1145-52.
33. Kubista B, Trieb K, Sevelda F, Toma C, Arrich F, Heffeter P, Elbling L, Sutterlüty H, Scotlandi K, Kotz R, Micksche M, Berger W. RhoA GTPase inactivation by statins induces osteosarcoma cell apoptosis by inhibiting p42/p44-MAPKs-Bcl-2 signaling independently of BMP-2 and cell differentiation. Cell Death Differ. 2006 Feb 10.
34. Alleva R, Benassi MS, Pazzaglia L, Tomasetti M, Gellert N, Borghi B, Neuzil J, Picci P. Alpha-tocopheryl succinate alters cell cycle distribution sensitising humanosteosarcoma cells to methotrexate-induced apoptosis. Cancer Lett. 2006 Feb 8;232(2):226-35.
35. Kang HJ, Jo C, Kwon JH, Son JH, An BJ, Byun MW. Antioxidant and cancer cell proliferation inhibition effect of citrus pectin-oligosaccharide prepared by irradiation. J Med Food. 2006 Fall;9(3):313-20.
36. Zou DM, Brewer M, Garcia F, Feugang JM, Wang J, Zang R, Liu H, Zou C. Cactus pear: a natural product in cancer chemoprevention. Nutr J. 2005 Sep 8;4:25.
37. Aires V, Adote S, Hichami A, Moutairou K, Boustani ES, Khan NA. Modulation of intracellular calcium concentrations and T cell activation by prickly pear polyphenols. Mol Cell Biochem. 2004 May;260(1-2):103-10.
38. Su CC, Lin JG, Li TM, Chung JG, Yang JS, Ip SW, Lin WC, Chen GW. Curcumin-induced apoptosis of human colon cancer colo 205 cells through the. production of ROS, Ca2+ and the activation of caspase-3. Anticancer Res. 2006 Nov-Dec;26(6B):4379-89.
39. Holmes FA, Madden T, Newman RA, Valero V, Theriault RL, Fraschini G, Walters RS, Booser DJ, Buzdar AU, Willey J, Hortobagyi GN. Sequence-dependent alteration of doxorubicin pharmacokinetics by paclitaxel in a phase I study of paclitaxel and doxorubicin in patients with metastatic breast cancer. J Clin Oncol. 1996 Oct;14(10): 2713-21.
40.王兵,王国俊,徐钧.氧化苦参碱对肿瘤诱导血管内皮细胞增殖的抑制作用[J].实用肿瘤杂志,2000,15(5):297-300.
41.周钢,杨述华,何宇.大蒜素诱导LM-8细胞凋亡及相关基因表达的实验研究.中国中医骨伤科杂志. 2007年08期.
42.杨迪生,解先宽,叶招明,陶惠民.腺病毒介导反义c-myc增强骨肉瘤细胞对顺铂化疗敏感性的实验研究.中华外科杂志. 2005 Jun 15;43(12):799-802.
43.高大新,张海燕,吕刚,李永军,黄涛,王岩峰. Survivin反义寡核苷酸联合顺铂治疗裸鼠荷骨肉瘤.中华实验外科杂志. 2005 Nov;22(8): 1003-4.
44. Van Valen F, Fulda S, Sch?fer KL, Truckenbrod B, Hotfilder M, Poremba C, Debatin KM, Winkelmann W. Selective and nonselective toxicity of TRAIL/Apo2L combinedwith chemotherapy in human bone tumour cells vs. normal human cells. Int J Cancer. 2003 Dec 20;107(6):929-40.
45. Yang DS, Miao XD, Ye ZM, Xu YS. Synergistic induction of apoptosis by the combination of TRAIL and low dose adriamycin in human osteosarcoma cell line U2OS. Zhonghua Zhong Liu Za Zhi. 2005 Oct;27(10):595-7.
46. Nardin A, Lefebvre ML, Labroquère K, Faure O, Abastado JP. Liposomal muramyl tripeptide phosphatidylethanolamine: Targeting and activating macrophages for adjuvant treatment of osteosarcoma. Curr Cancer Drug Targets. 2006 Mar;6(2):123-33.
47. Sherr CJ. Principles of tumor suppression. Cell. 2004 Jan 23;116(2):235-46.
48. McCormick F. Cancer gene therapy: Fringe or cutting edge? Nat Rev Cancer,2001, 1:130-41.
49. Van Meir EG, Polverini PJ, Chazin VR, Su Huang HJ, de Tribolet N, Cavenee WK. Release of an inhibitor of angiogenesis upon induction of wild type p53 expression in glioblastoma cells. Nat Genet. 1994 Oct;8(2):171-6.
50. Takaoka A, Hayakawa S, Yanai H, Stoiber D, Negishi H, Kikuchi H, Sasaki S, Imai K, Shibue T, Honda K, Taniguchi T. Integration of interferon-alpha/beta signalling to p53 responses in tumour suppression and antiviral defence. Nature 2003;424:516–523.
51. Tataria M, Quarto N, Longaker MT, Sylvester KG. Absence of the p53 tumor suppressor gene promotes osteogenesis in mesenchymal stem cells. J Pediatr Surg. 2006 Apr;41(4): 624-32; discussion 624-32.
52. Graat HC, Wuisman PI, van Beusechem VW, Carette JE, Gerritsen WR, Bras J, Schaap GR, Kaspers GJ, Ogose A, Gu W, Kawashima H, Hotta T. Coxsackievirus and adenovirus receptor expression on primary osteosarcoma specimens and implications for gene therapy with recombinant adenoviruses. Clin Cancer Res. 2005 Mar 15;11(6):2445-7; author reply 2447-8.
53. Ternovoi VV, Curiel DT, Smith BF, Siegal GP. Adenovirus-mediated p53 tumor suppressor gene therapy of osteosarcoma. Lab Invest. 2006 Aug;86(8):748-66. Epub 2006 Jun 5. Review.
54. Kwon KB, Yoo SJ, Ryu DG, Yang JY, Rho HW, Kim JS, Park JW, Kim HR, Park BH. Induction of apoptosis by diallyl disulfide through activation of caspase-3 in humanleukemia HL-60 cells. Biochem Pharmacol. 2002 Jan 1;63(1):41-7.
55. Green R, Reed C. Mitochondria and apoptosis. Science, 1998;281(6);1309-1312
56. Kaseta MK, Khaldi L, Gomatos IP, Tzagarakis GP, Alevizos L, Leandros E, Papagelopoulos PJ, Soucacos PN. Kaseta MK, Khaldi L, Gomatos IP, Tzagarakis GP, Alevizos L, Leandros E, Papagelopoulos PJ, Soucacos PN. J Surg Oncol. 2008 Mar 1;97(3):259-66.