Albaconol的抗肿瘤活性及对拓扑异构酶活性和血管生成的影响
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摘要
目的 研究地花菌(Albatrellus Confluens)提取物Albaconol的抗肿瘤活性，并进一步探讨Albaconol对DNA拓扑异构酶活性和新生血管生成的影响。方法 以人白血病细胞株K562、人乳腺癌细胞株Bcap-37、人胃腺癌细胞株BGC-823和人非小细胞肺癌细胞株A549为模型，采用MTT法测试Albaconol对肿瘤细胞增殖的影响；以小鼠肉瘤S180和小鼠肝癌H22为模型，检测Albaconol静脉给药对小鼠移植性肿瘤生长的影响；以pBR322 DNA为底物，采用琼脂糖凝胶电泳法测定Albaconol对肿瘤细胞DNA拓扑异构酶活性的影响；以HMEC细胞增殖、迁移和小管形成为模型，测试Albaconol对血管生成的作用。结果 Albaconol对K562、Bcap-37、BGC．823、A549的半数抑制浓度(IC_(50))分别为2.42+1.77、1.88±1.41、1.04+0.64、1.18±1.10μg/ml，均小于10μg／ml；Albaconol低、中、高剂量组(0.87、1.73、3.46mg／kg)对S180的抑瘤率分别为28.2％、43.2％、47.4％(P＜0.01)，对H22的抑瘤率分别为15.6％(P＞0.05)、22.4％(P＜0.05)、37.8％(P＜0.01)，且呈现出较好的剂量依赖性；Albaconol能明显影响DNA拓扑异构酶Ⅱ的活性，表现为促进其介导的DNA解旋或断裂，并抑制其介导的DNA再连接反应，但对DNA拓扑异构酶Ⅰ的活性几无影响；Albaconol能明显抑制HMEC细胞增殖、迁移和小管形成，且呈现出剂量依赖关系。结论 Albaconol具有较强的体内、外抗肿瘤活性；Albaconol能选择性地影响DNA拓扑异构酶Π 的活性，DNA拓扑异构酶Π是其抗肿瘤作用的细胞内靶点之一；Albaconol能明显抑制HMEC细胞增殖、迁移和小管形成，抗血管生成是其抗肿瘤作用的机制之一。
To study the antitumor activity of Albaconol from Albatrellus confluens, the effect on DNA topoisomerase (TOPO) and angiogenesis activity in vitro. METHODSrThe cytotoxicity of Albaconol on human leukemia K562 , breast carcinoma Bcap-37, gastric adenocarcinoma BGC-823 and lung adenocarcinoma A549 cell lines were determined by MTT assay and the results were expressed as IC50 values; The inhibitory effect of Albaconol on tumor growth was observed by the models of implanted sarcoma 180 (SI80) and hepatoma 22 (H22) in mice and the results were expressed as inhibitory rate of tumor growth; The effect of Albaconol on DNA topoisomerase activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate; The antiangiogenesis activity of Albaconol on human microvascular endothelial cells (HMECs) was investigated by HMECs growth inhibition assay ( MTT assay ) , Wounding Migration Assay and Matrigel Tube Formation Assay. RESULTS:The results showed that the IC50 values of Albaconol on K562, Bc
ap-37 > BGC-823 and A549 cell lines were 2.42+1.77, 1.8811.4K 1.0410.64 and 1.1811.10 ng/ml, respectively; The inhibitory rates of tumor growth at doses of 0.87, 1.74, 3.48 mg/kg of Albaconol on S180 and H22were 28.2 %, 43.2 %, 47.4 % and 15.6 %, 22.4 %, 37.8 %, respectively; Albaconol influenced obviously the activity of DNA TOPO II, stimulated DNA cleavage and inhibited DNA reunion mediated by TOPO II; Albaconol inhibited the proliferation of HMECs, the IC50 values were 17.52?.13 g/ml. Furthermore, Albaconol strongly inhibited the migration ability of HMECs induced by hEGF in a dose-dependent manner and greatly inhibited capillary-like networks development of HMECs. CONCLUSION:Albaconol exhibited significant antitumor activity in vitro and in vivo;
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Albaconol targeted specifically DNA TOPO II , which is one of the mechanisms of its antitumor action: Albaconol has strong angiogenic effect on three important sequential angiogenic cascades and antiangiogenesis is one of the mechanisms of its antitumor action.
To study the antitumor activity of Albaconol from Albatrellus confluens, the effect on DNA topoisomerase (TOPO) and angiogenesis activity in vitro. METHODSrThe cytotoxicity of Albaconol on human leukemia K562 , breast carcinoma Bcap-37, gastric adenocarcinoma BGC-823 and lung adenocarcinoma A549 cell lines were determined by MTT assay and the results were expressed as IC50 values; The inhibitory effect of Albaconol on tumor growth was observed by the models of implanted sarcoma 180 (SI80) and hepatoma 22 (H22) in mice and the results were expressed as inhibitory rate of tumor growth; The effect of Albaconol on DNA topoisomerase activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate; The antiangiogenesis activity of Albaconol on human microvascular endothelial cells (HMECs) was investigated by HMECs growth inhibition assay ( MTT assay ) , Wounding Migration Assay and Matrigel Tube Formation Assay. RESULTS:The results showed that the IC50 values of Albaconol on K562, Bc
ap-37 > BGC-823 and A549 cell lines were 2.42+1.77, 1.8811.4K 1.0410.64 and 1.1811.10 ng/ml, respectively; The inhibitory rates of tumor growth at doses of 0.87, 1.74, 3.48 mg/kg of Albaconol on S180 and H22were 28.2 %, 43.2 %, 47.4 % and 15.6 %, 22.4 %, 37.8 %, respectively; Albaconol influenced obviously the activity of DNA TOPO II, stimulated DNA cleavage and inhibited DNA reunion mediated by TOPO II; Albaconol inhibited the proliferation of HMECs, the IC50 values were 17.52?.13 g/ml. Furthermore, Albaconol strongly inhibited the migration ability of HMECs induced by hEGF in a dose-dependent manner and greatly inhibited capillary-like networks development of HMECs. CONCLUSION:Albaconol exhibited significant antitumor activity in vitro and in vivo;
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Albaconol targeted specifically DNA TOPO II , which is one of the mechanisms of its antitumor action: Albaconol has strong angiogenic effect on three important sequential angiogenic cascades and antiangiogenesis is one of the mechanisms of its antitumor action.
引文
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[3] Wane LG, Liu XM, Ji XJ. Determination of DNA topoisomerase Ⅱ activity from L_(1210) cells——a target for screening antitumor agent.Acta Pharmacol Sin. 1991, 12:108
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[9] Yang WM, Liu JK, Qing C, et al. Albaconol from the Mushroom Albatrellus confluens induces contraction and desensitization in guinea pig trachea. Planta Med, 2003; 69:715-9
[10] Sullivan DM, Glisson BS, Hodges PK, et al. Proliferation dependence of topoisomerase Ⅱ mediated drug action. Biochemistry. 1986; 25:2248-56
[11] 周建军，乐秀芳，韩家娴等．评价抗癌物质活性的改良MTT方法．中国医药工业杂志，1993，24(10)：455-457
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[13] 卿晨，纪舒昱，王蕾．青山抗癌注射液抑制小鼠肿瘤生长的实验研究．昆明医学院学报，2003；24(2)：18～9
[14] 蒙凌华，蒋超，刘兆乾．螺旋藻提取物对DNA拓扑异构酶活性的抑制及对DNA的直接影响．癌症，2000，19(8)：768-771
[15] Mi-Sook L, Eun-Joung M, Sae-Won L,et al. Angiogenic activity of pyruvic acid in vivo and in vitro angiogenesis models. Cancer Research .2001 ;61:3290-3293.
[16] 沈关心，等主编．现代免疫学实验技术．武汉：湖北科学技术出版社，2001，505
[17] J.Folkman. How is blood vessel growth regulated in normal andneoplastic tissue. Cancer Res, 1986; 46: 467-473.
[18] Corinne L, Reimer, Naoki A, et al. Antieoplastic effects of chemotherapeutic agents are potentiated by NM-3, an inhibitor of angiogenesis. Cancer Research .2002; 62:789-795.
[19] 李宗金，韩忠朝．血管发生和血管新生的体外模型．中国微循环．2003；7(1)：52-55
[20] 新药(西药)临床前研究指导原则汇编．中华人民共和国卫生部药政局，1993，137-139
[21] Pourquier P, Pommier Y. topoisomerases Ⅰ: new targets for the treatment of cancer and mechanisms of resistance. Bull Cancer, 1998;9:5～10.
[22] Peng Y, Bao DY, Wang LG, Liu XM, Ji XJ. Effects of siwenmycin on DNA topoisomerases mediated strand cleavage and relaxation in vitro. J Chin Pharm Sci. 1994; 3: 120-5
[23] Eckhardt SG.Angiogenesis inhibitors as cancer therapy. Hospital practice, 1999; 34(1):63-69.
[24] Robert J, Schatieman GC. Angiogenesis: new insights and therapeutic potential. The Anatomical Record(new anat),2000; 261:126-135.
[25] Gnffioen AW, Barendsz-Janson AF, Mayo KH, et al. Angiogenesis, a target for tumor therapy. J Lab Clin Med. 1998; 132:363-368.
[26] Kuwano M, Fukushi J, Okamoto M, et al. Angiogenesis factors.Intern Med, 2001 ;40(7):565-72.
[27] Gazdar AF, Minna JD. Angiogenesis and the Multistage Development of Lung Cancers. Clinical Cancer Research, 2000; 6:1611-1612.
[28] Bergers G., Javaherian K, Lo K. M, et al. Effects of angiogenesis inhibitors on multistage carcinogenesis in mice. Science (Washington DC), 1999; 284: 808-812.
[2] Osheroff N Biochemical basis for the interactions of type Ⅰ and type Ⅱ topoisomerases. with DNA Pharmacol Ther. 1989; 41 : 223-41
[3] Wane LG, Liu XM, Ji XJ. Determination of DNA topoisomerase Ⅱ activity from L_(1210) cells——a target for screening antitumor agent.Acta Pharmacol Sin. 1991, 12:108
[4] Risau W. Mechanisms of angiogenesis. Nature, 1997;Apr 17;386:671-674
[5] Folkman J. Angoigenesis in cancer, vascular, reumatoid and other disease[J].Nature Med,1995, 1:27-31.
[6] Folkrnan J.Tumor angiogenesis:therapeutic implications.N.Engl.J.Med, 1971,285:1182 - 1186
[7] Folkman J. J Natl Cancer Inst,2000;92(2):94
[8] Ding ZH, Dong ZJ, Liu JK. Albaconol, a novel prenylated resorcinol (= Benzene-1,3-diol) from Basidiomycetes Albatrellus confluens. Helv Chim Acta, 2001; 84:259-62
[9] Yang WM, Liu JK, Qing C, et al. Albaconol from the Mushroom Albatrellus confluens induces contraction and desensitization in guinea pig trachea. Planta Med, 2003; 69:715-9
[10] Sullivan DM, Glisson BS, Hodges PK, et al. Proliferation dependence of topoisomerase Ⅱ mediated drug action. Biochemistry. 1986; 25:2248-56
[11] 周建军，乐秀芳，韩家娴等．评价抗癌物质活性的改良MTT方法．中国医药工业杂志，1993，24(10)：455-457
[12] 徐叔云，卞如濂，陈修主编．药理实验方法学，第三版，北京：人民卫生出版社，2002：1757～79
[13] 卿晨，纪舒昱，王蕾．青山抗癌注射液抑制小鼠肿瘤生长的实验研究．昆明医学院学报，2003；24(2)：18～9
[14] 蒙凌华，蒋超，刘兆乾．螺旋藻提取物对DNA拓扑异构酶活性的抑制及对DNA的直接影响．癌症，2000，19(8)：768-771
[15] Mi-Sook L, Eun-Joung M, Sae-Won L,et al. Angiogenic activity of pyruvic acid in vivo and in vitro angiogenesis models. Cancer Research .2001 ;61:3290-3293.
[16] 沈关心，等主编．现代免疫学实验技术．武汉：湖北科学技术出版社，2001，505
[17] J.Folkman. How is blood vessel growth regulated in normal andneoplastic tissue. Cancer Res, 1986; 46: 467-473.
[18] Corinne L, Reimer, Naoki A, et al. Antieoplastic effects of chemotherapeutic agents are potentiated by NM-3, an inhibitor of angiogenesis. Cancer Research .2002; 62:789-795.
[19] 李宗金，韩忠朝．血管发生和血管新生的体外模型．中国微循环．2003；7(1)：52-55
[20] 新药(西药)临床前研究指导原则汇编．中华人民共和国卫生部药政局，1993，137-139
[21] Pourquier P, Pommier Y. topoisomerases Ⅰ: new targets for the treatment of cancer and mechanisms of resistance. Bull Cancer, 1998;9:5～10.
[22] Peng Y, Bao DY, Wang LG, Liu XM, Ji XJ. Effects of siwenmycin on DNA topoisomerases mediated strand cleavage and relaxation in vitro. J Chin Pharm Sci. 1994; 3: 120-5
[23] Eckhardt SG.Angiogenesis inhibitors as cancer therapy. Hospital practice, 1999; 34(1):63-69.
[24] Robert J, Schatieman GC. Angiogenesis: new insights and therapeutic potential. The Anatomical Record(new anat),2000; 261:126-135.
[25] Gnffioen AW, Barendsz-Janson AF, Mayo KH, et al. Angiogenesis, a target for tumor therapy. J Lab Clin Med. 1998; 132:363-368.
[26] Kuwano M, Fukushi J, Okamoto M, et al. Angiogenesis factors.Intern Med, 2001 ;40(7):565-72.
[27] Gazdar AF, Minna JD. Angiogenesis and the Multistage Development of Lung Cancers. Clinical Cancer Research, 2000; 6:1611-1612.
[28] Bergers G., Javaherian K, Lo K. M, et al. Effects of angiogenesis inhibitors on multistage carcinogenesis in mice. Science (Washington DC), 1999; 284: 808-812.