扶正肝康颗粒对H_（22）荷瘤ICR小鼠肿瘤和免疫功能影响及毒性研究
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摘要
本研究通过建立ICR小鼠Hep-A-22(以下称为H22)肝癌移植性实体肿瘤模型,对扶正肝康颗粒对H22荷瘤ICR小鼠肿瘤、抗肿瘤免疫功能、肝肾毒性进行了初步考察。结果表明,扶正肝康颗粒对H22荷瘤ICR小鼠有一定体内抑瘤作用,使H22实体瘤异型性有所改善;对H22荷瘤ICR小鼠抗肿瘤免疫功能无显著影响,胸腺、脾脏重量和脏器指数、血清TNF-α、IL-2无显著改变,血清IFN-γ显著降低;对H22荷瘤ICR小鼠肝脏无明显毒性,肝脏重量和脏器指数无显著改变,血清TP、ALB、GLO、ALT、ALP以及肝组织tSOD、GSH-PX无显著变化;中剂量表现为H22荷瘤ICR小鼠血清BUN显著下降。综上认为,扶正肝康颗粒对肝脏肿瘤未见有明显抑制作用,未发现有促进荷瘤小鼠免疫功能作用,未见对荷瘤小鼠肝功能有显著影响。本次试验结果表明扶正肝康颗粒在肝脏肿瘤研究方面结果不甚理想,但仍为该药物在肝病相关领域的应用提供了一部分基础性数据,对该药物在有效领域的研究提供一个对比参考。
Objective:
Fuzhenggankang Granule, an compound recipe, is determined as an assistant drug for the therapy of liver diseases. The preliminary study found that Fuzhenggankang Granule had an efficacy on liver cirrhotic ascites based on the rat model of that disease induced by CCL4 compositing gavage. In order to investigate whether this drug had some effects on the other types of common liver diseases besides the liver cirrhotic ascites, the ICR mouse was used in this study and also the mouse hepatoma cell line H22 to establish the H22-bearing ICR mouse model, and then we observed the effests of Fuzhenggankang Granule in the aspects of the tumor , the immune function and the function of other organs such as the liver and the kidney, looking forward to expanding the awareness and the therapeutic applications of the drug.
Methods:
The resuscitated H22 cell line was recovered in ICR mouse peritoneal cavity, with 1.0×108 cells per milliliter as the fina use concentration and then inoculated to ICR mouse by axillary subcutaneous transplantation with 0.2mL cell suspension of each model mouse to construct the hepatoma solid tumor model. The model mice were sent to positive control, negaive control and the drug assay groups randomly. The final concentration of the positive control drug CTX (powder) dissoved in the normal saline was 4 g·L-1, and its daily dose was 0.02 g for each kilogram body weight by intraperitoneal injection once a day, equal to the daily volume of 0.05 mL CTX solution for 10 g body weight. As the negative control, normal saline was given 0.2 mL for 10 g body weight once a day as the daily dose by gavage. The high, middle and low doses of Fuzhenggankang Granule are respectively 7.20 g, 3.60 g and 1.20 g for each kilogram body weight by gavage once a day as the daily doses, and the preparing concentrations were 0.36 g·mL-1, 0.12 g·mL-1 and 0.06 g·mL-1 equal to the daily volume of 0.2 mL for 10 g body weight respectively. Fuzhenggankang Granule, positive control and negative control drugs were began to given respectively at the following day after the H22 inoculation, lasting 10 days continually. After 24h of the latest drug administration, the model mice were executed and the specimens were collected. The drug control(middle) group also used the ICR mouse and were raised and administrated with the middle dosage at equal pace with others, but no H22 cell inoculation. During the 10 days of the drug administration, the body weight of each model mouse was measured as a reference to adjust the drug administration volume to it. The blood was collected by enucleation of eyeball ,and after separation the serum was stored at -80℃for the detection of TP, ALB, GLO, ALT, ALP, BUN, TNF-α, IFN-γand IL-2. The weights of fresh tissues including thymus, spleen, liver and solid tumor were measured and preserved in 4 % formaldehyde for paraffin section preparation and the further Hematoxylin-Eosin staining. Part of the fresh liver tissues was preserved at -80℃for the tissue homogenate preparation and the further detection of tSOD, GSH-PX in liver.
Results:
Compared with the negative control, the tumor inhibition rates of Fuzhenggankang Granule had a relatively better inhibition rate(24.5 %) in the middle dose(3.60 g·kg-1BW·d-1)of Fuzhenggankang Granule, but no statistical significance. Compared with the negative control, Hematoxylin-Eosin staining of the H22 solid tumor showed that both the high and the middle drug groups had remissions of atypia to some extent, revealed the clearer intercellular connections and cell contours, as well as the decrease staining of nuclei and the improvement in nucleo-cytoplasmic ratio. Compared with the negative control, the drug had no significant effects on the weight of thymus and the spleen as well as their organ indexes and histomorphology. Compared with the negative control, the drug had no significant effects on the levels of TNF-α, IL-2, but with a decrease of IFN-γin serum. Compared with the negative control, the drug had no significant effect on the ponderal growth. Compared with the negative control, the drug had no significant effect on the liver in terms of the organ weight, the organ index and the histomorphology. Compared with the negative control, there were no statistical changes in the aspects of TP, ALB, GLO and ALB/GLO, showing the protein synthesis function of liver. The ALT, ALP activities in the serm, the tSOD, GSH-PX activities in the liver had no significant changes as well. Compared with the negative control, the BUN in serum had significant decrease in the drug middle group.
Conclusions:
1. Fuzhenggankang Granule had a certain tumor inhibitory effect on the solid tumor in H22-bearing ICR mouse model and the atypia of the solid tumor eased at the same time. But the inhibitory rate had no statistical significance.
2. Fuzhenggankang Granule had no significant effect on the anti-tumor immune function in H22-bearing ICR mouse model. The weight and the organ index of thymus and spleen had no significant changes, and also the levels of TNF-αand IL-2 in serum. But the IFN-γhad a significant decrease.
3. Fuzhenggankang Granule had no significant toxic effect on the liver of H22-bearing ICR mouse model in the aspects of the organ weight and index, and the activities of the hepatic enzymes such as ALT, ALP in serum and tSOD, GSH-PX in tissue.
4. Fuzhenggankang Granule showed out the significant decrease of BUN in serum in the middle drug dose.
Objective:
Fuzhenggankang Granule, an compound recipe, is determined as an assistant drug for the therapy of liver diseases. The preliminary study found that Fuzhenggankang Granule had an efficacy on liver cirrhotic ascites based on the rat model of that disease induced by CCL4 compositing gavage. In order to investigate whether this drug had some effects on the other types of common liver diseases besides the liver cirrhotic ascites, the ICR mouse was used in this study and also the mouse hepatoma cell line H22 to establish the H22-bearing ICR mouse model, and then we observed the effests of Fuzhenggankang Granule in the aspects of the tumor , the immune function and the function of other organs such as the liver and the kidney, looking forward to expanding the awareness and the therapeutic applications of the drug.
Methods:
The resuscitated H22 cell line was recovered in ICR mouse peritoneal cavity, with 1.0×108 cells per milliliter as the fina use concentration and then inoculated to ICR mouse by axillary subcutaneous transplantation with 0.2mL cell suspension of each model mouse to construct the hepatoma solid tumor model. The model mice were sent to positive control, negaive control and the drug assay groups randomly. The final concentration of the positive control drug CTX (powder) dissoved in the normal saline was 4 g·L-1, and its daily dose was 0.02 g for each kilogram body weight by intraperitoneal injection once a day, equal to the daily volume of 0.05 mL CTX solution for 10 g body weight. As the negative control, normal saline was given 0.2 mL for 10 g body weight once a day as the daily dose by gavage. The high, middle and low doses of Fuzhenggankang Granule are respectively 7.20 g, 3.60 g and 1.20 g for each kilogram body weight by gavage once a day as the daily doses, and the preparing concentrations were 0.36 g·mL-1, 0.12 g·mL-1 and 0.06 g·mL-1 equal to the daily volume of 0.2 mL for 10 g body weight respectively. Fuzhenggankang Granule, positive control and negative control drugs were began to given respectively at the following day after the H22 inoculation, lasting 10 days continually. After 24h of the latest drug administration, the model mice were executed and the specimens were collected. The drug control(middle) group also used the ICR mouse and were raised and administrated with the middle dosage at equal pace with others, but no H22 cell inoculation. During the 10 days of the drug administration, the body weight of each model mouse was measured as a reference to adjust the drug administration volume to it. The blood was collected by enucleation of eyeball ,and after separation the serum was stored at -80℃for the detection of TP, ALB, GLO, ALT, ALP, BUN, TNF-α, IFN-γand IL-2. The weights of fresh tissues including thymus, spleen, liver and solid tumor were measured and preserved in 4 % formaldehyde for paraffin section preparation and the further Hematoxylin-Eosin staining. Part of the fresh liver tissues was preserved at -80℃for the tissue homogenate preparation and the further detection of tSOD, GSH-PX in liver.
Results:
Compared with the negative control, the tumor inhibition rates of Fuzhenggankang Granule had a relatively better inhibition rate(24.5 %) in the middle dose(3.60 g·kg-1BW·d-1)of Fuzhenggankang Granule, but no statistical significance. Compared with the negative control, Hematoxylin-Eosin staining of the H22 solid tumor showed that both the high and the middle drug groups had remissions of atypia to some extent, revealed the clearer intercellular connections and cell contours, as well as the decrease staining of nuclei and the improvement in nucleo-cytoplasmic ratio. Compared with the negative control, the drug had no significant effects on the weight of thymus and the spleen as well as their organ indexes and histomorphology. Compared with the negative control, the drug had no significant effects on the levels of TNF-α, IL-2, but with a decrease of IFN-γin serum. Compared with the negative control, the drug had no significant effect on the ponderal growth. Compared with the negative control, the drug had no significant effect on the liver in terms of the organ weight, the organ index and the histomorphology. Compared with the negative control, there were no statistical changes in the aspects of TP, ALB, GLO and ALB/GLO, showing the protein synthesis function of liver. The ALT, ALP activities in the serm, the tSOD, GSH-PX activities in the liver had no significant changes as well. Compared with the negative control, the BUN in serum had significant decrease in the drug middle group.
Conclusions:
1. Fuzhenggankang Granule had a certain tumor inhibitory effect on the solid tumor in H22-bearing ICR mouse model and the atypia of the solid tumor eased at the same time. But the inhibitory rate had no statistical significance.
2. Fuzhenggankang Granule had no significant effect on the anti-tumor immune function in H22-bearing ICR mouse model. The weight and the organ index of thymus and spleen had no significant changes, and also the levels of TNF-αand IL-2 in serum. But the IFN-γhad a significant decrease.
3. Fuzhenggankang Granule had no significant toxic effect on the liver of H22-bearing ICR mouse model in the aspects of the organ weight and index, and the activities of the hepatic enzymes such as ALT, ALP in serum and tSOD, GSH-PX in tissue.
4. Fuzhenggankang Granule showed out the significant decrease of BUN in serum in the middle drug dose.
引文
[1] Ramadori G, Moriconi F, Malik I, et al. Physiology and pathophysiology of liver inflammation, damage and repair[J]. J Physiol Pharmacol, 2008, 59Suppl 1:107-117.
[2] El-Serag HB, Rudolph KL. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis[J]. Gastroenterology, 2007, 132(7):2557-2576.
[3] Motola-Kuba D, Zamora-Valdés D, Uribe M, et al. Hepatocellular carcinoma. An overview[J]. Ann Hepatol, 2006, 5(1):16-24.
[4] Luo RH, Zhao ZX, Zhou XY, et al. Risk factors for primary liver carcinoma in Chinese population[J]. World J Gastroenterol, 2005, 11(28):4431-4434.
[5] Bosch FX, Ribes J, Díaz M, et al. Primary liver cancer: worldwide incidence and trends[J]. Gastroenterology, 2004, 127(5 Suppl 1):S5-S16.
[6] Marrero CR, Marrero JA. Viral hepatitis and hepatocellular carcinoma[J]. Arch Med Res, 2007, 38(6):612-620.
[7] Marrero JA. Hepatocellular carcinoma[J]. Curr Opin Gastroenterol, 2005, 21(3):308-312.
[8] Gomaa AI, Khan SA, Toledano MB, et al. Hepatocellular carcinoma: epidemiology, risk factors and pathogenesis[J]. World J Gastroenterol, 2008, 14(27):4300-4308.
[9]乔凤娟,梁睿,牛凤兰,等.东北菱对鼠肝癌H22细胞凋亡及细胞周期进程的影响[J].中国卫生工程学, 2006, 5(6):364-365.
[10]牛凤兰,董威严,乔凤娟.菱角提取物对荷瘤小鼠免疫功能的影响[J].食品科学, 2006, 27(7):233-235.
[11]吕喆,龚守良,牛凤兰,等.菱角纯化物三羟基苯甲酸二聚体对肝癌SMMC-7721细胞凋亡及细胞周期进程的影响[J].吉林大学学报(医学版), 2006, 32(5):788-790.
[12] Niu FL, Wang XD, Wang YL, et al. Trihydroxybenzoic acid dimer-induced apoptosis effect in vitro[J]. Chem Res Chinese U, 2005, 21(4):463-467.
[13]赵宏艳,张皖东,吕诚,等.灵芝提取物对H22荷瘤小鼠抗肿瘤作用的实验研究[J].中国中药杂志, 2008, 33(7):842-845.
[14]宁安红,曹婧,黄敏,等.灵芝多糖对小鼠肿瘤的抑制作用及免疫指标的观察[J].肿瘤研究与临床, 2004, 16(3):147-149.
[15]彭玉娜,谭获,王颖超,等.灵芝孢子粉对兔移植性肿瘤生存期及外周血的影响[J].时珍国医国药, 2008, 19(7):1737-1738.
[16]李颖博,李宇华,王瑞,等.灵芝多糖对肿瘤细胞与内皮细胞相互作用的影响[J].中国药理学通报, 2008, 24(2):250-253.
[17] Fujii E, Suzuki M, Matsubara K, et al. Establishment and characterization of in vivo human tumor models in the NOD/SCID/gamma(c)(null) mouse[J]. Pathol Int, 2008, 58(9):559-567.
[18] Kerbel RS. Human tumor xenografts as predictive preclinical models for anticancer drug activity in humans: better than commonly perceived-but they can be improved[J]. Cancer Biol Ther, 2003, 2(4 Suppl 1):S134-139.
[19] Uckun FM. Severe combined immunodeficient mouse models of human leukemia[J]. Blood, 1996, 88(4):1135-1146.
[20] Sausville EA, Burger AM. Contributions of human tumor xenografts to anticancer drug development[J]. Cancer Res, 2006, 66(7):3351-3354, discussion 3354.
[21] Teicher BA. Tumor models for efficacy determination[J]. Mol Cancer Ther, 2006, 5(10):2435-2443.
[22] Carver BS, Pandolfi PP. Mouse modeling in oncologic preclinical and translational research[J]. Clin Cancer Res, 2006, 12(18):5305-5311.
[23] Khanna C, Hunter K. Modeling metastasis in vivo[J]. Carcinogenesis, 2005, 26(3):513-523.
[24] Ohsugi T, Yamaguchi K, Kumasaka T, et al. Rapid tumor death model for evaluation of new therapeutic agents for adult T-cell leukemia[J]. Lab Invest, 2004, 84(2):263-266.
[25] Loisel S, Ster KL, Quintin-Roue I, et al. Establishment of a novel human B-CLL-like xenograft model in nude mouse[J]. Leuk Res, 2005, 29(11):1347-1352.
[26] Macor P, Secco E, Zorzet S, et al. An update on the xenograft and mouse models suitable for investigating new therapeutic compounds for the treatment of B-cell malignancies[J]. Curr Pharm Des, 2008, 14(21):2023-2039.
[27] Reuther T, Kübler AC, Staff CJ, et al. The RAG 2 mouse model for xenografted human oral squamous cell carcinoma[J]. Contemp Top Lab Anim Sci, 2002, 41(2):31-35.
[28] Fichtner I, Rolff J, Soong R, et al. Establishment of patient-derived non-small cell lung cancer xenografts as models for the identification of predictive biomarkers[J]. Clin Cancer Res, 2008, 14(20):6456-6468.
[29] Viaggi M, Dagrosa MA, Belli C, et al. A new animal model for human undifferentiated thyroid carcinoma[J]. Thyroid, 2003, 13(6):529-536.
[30] Wang Y, Sudilovsky D, Zhang B, et al. A human prostatic epithelial model of hormonal carcinogenesis[J]. Cancer Res, 2001, 61(16):6064-6072.
[31] Ke N, Zhou D, Chatterton JE, et al. A new inducible RNAi xenograft model for assessing the staged tumor response to mTOR silencing[J]. Exp Cell Res, 2006, 312(15):2726-2734.
[32] Elkas JC, Baldwin RL, Pegram M, et al. A human ovarian carcinoma murine xenograft model useful for preclinical trials[J]. Gynecol Oncol, 2002, 87(2):200-206.
[33] Orazine CI, Hincapie M, Hancock WS, et al. A proteomic analysis of the plasma glycoproteins of a MCF-7 mouse xenograft: a model system for the detection of tumor markers[J]. J Proteome Res, 2008, 7(4):1542-1554.
[34] Henriksson KC, Almgren MA, Thurlow R, et al. A fluorescent orthotopic mouse model for reliable measurement and genetic modulation of human neuroblastoma metastasis[J]. Clin Exp Metastasis, 2004, 21(6):563-570.
[35] De Velasco MA, Tanaka M, Anai S, et al. GFP image analysis in the mouse orthotopic bladder cancer model[J]. Oncol Rep, 2008, 20(3):543-547.
[36] Cullinane C, Dorow DS, Kansara M, et al. An in vivo tumor model exploiting metabolic response as a biomarker for targeted drug development[J]. Cancer Res, 2005, 65(21):9633-9636.
[37] Haldar M, Randall RL, Capecchi MR. Synovial sarcoma: from genetics to genetic-based animal modeling[J]. Clin Orthop Relat Res, 2008 Sep;466(9):2156-2167. Epub 2008 Jun 18.
[38] Stell A, Biserni A, Della Torre S, et al. Cancer modeling: modern imaging applications in the generation of novel animal model systems to study cancerprogression and therapy[J]. Int J Biochem Cell Biol, 2007, 39(7-8):1288-1296.
[39] Malik S, Balkwill FR. Tumour necrosis factor[J]. Br Med J (Clin Res Ed), 1988, 296(6631):1214.
[40] Spriggs D, Imamura K, Rodriguez C, et al. Induction of tumor necrosis factor expression and resistance in a human breast tumor cell line[J]. Proc Natl Acad Sci U S A, 1987, 84(18):6563-6566.
[41] Candolfi M, Jaita G, Pisera D, et al. Adenoviral vectors encoding tumor necrosis factor-alpha and FasL induce apoptosis of normal and tumoral anterior pituitary cells[J]. J Endocrinol, 2006, 189(3):681-690.
[42] Goodsell DS. The molecular perspective: tumor necrosis factor[J]. Oncologist, 2006, 11(1):83-84.
[43] Verhoef C, de Wilt JH, Grünhagen DJ, et al. Isolated limb perfusion with melphalan and TNF-alpha in the treatment of extremity sarcoma[J]. Curr Treat Options Oncol, 2007, 8(6):417-427.
[44] van Horssen R, Ten Hagen TL, Eggermont AM. TNF-alpha in cancer treatment: molecular insights, antitumor effects, and clinical utility[J]. Oncologist, 2006, 11(4):397-408.
[45] Bellone M, Mondino A, Corti A. Vascular targeting, chemotherapy and active immunotherapy: teaming up to attack cancer[J]. Trends Immunol, 2008, 29(5):235-241.
[46] Williams GM. Antitumor necrosis factor-alpha therapy and potential cancer inhibition[J]. Eur J Cancer Prev, 2008, 17(2):169-177.
[47] Suganuma M, Kuzuhara T, Yamaguchi K, et al. Carcinogenic role of tumor necrosis factor-alpha inducing protein of Helicobacter pylori in human stomach[J]. J Biochem Mol Biol, 2006, 39(1):1-8.
[48] Alexander HR Jr, Bartlett DL, Libutti SK. Current status of isolated hepatic perfusion with or without tumor necrosis factor for the treatment of unresectable cancers confined to liver[J]. Oncologist, 2000, 5(5):416-424.
[49] Bubeník J. Interleukin-2 therapy of cancer[J]. Folia Biol (Praha), 2004, 50(3-4):120-130.
[50] Rosenberg SA. The development of new immunotherapies for the treatment ofcancer using interleukin-2[J]. Ann Surg, 1988, 208(2):121-135.
[51] Uchiyama T. Interleukin-2 and its receptor in adult T cell leukemia[J]. Jpn J Med, 1991, 30(6):602-606
[52] Overwijk WW, Theoret MR, Restifo NP. The future of interleukin-2: enhancing therapeutic anticancer vaccines[J]. Cancer J Sci Am, 2000, 6 Suppl 1:S76-80.
[53] Atkins MB, Regan M, McDermott D. Update on the role of interleukin 2 and other cytokines in the treatment of patients with stage IV renal carcinoma[J]. Clin Cancer Res, 2004, 10(18 Pt 2):6342S-6346S.
[54] McDermott DF. Update on the application of interleukin-2 in the treatment of renal cell carcinoma[J]. Clin Cancer Res, 2007, 13(2 Pt 2):716s-720s.
[55] Adolf GR. Structure and effects of interferon-gamma[J]. Oncology, 1985, 42 Suppl 1:33-40.
[56] Blankenstein T, Qin Z. The role of IFN-gamma in tumor transplantation immunity and inhibition of chemical carcinogenesis[J]. Curr Opin Immunol, 2003, 15(2):148-154.
[57] Qin Z, Schwartzkopff J, Pradera F, et al. A critical requirement of interferon gamma-mediated angiostasis for tumor rejection by CD8+ T cells[J]. Cancer Res, 2003, 63(14):4095-4100.
[58] Merchant MS, Yang X, Melchionda F, et al. Interferon gamma enhances the effectiveness of tumor necrosis factor-related apoptosis-inducing ligand receptor agonists in a xenograft model of Ewing's sarcoma[J]. Cancer Res, 2004, 64(22):8349-8356.
[59] Zhang B, Karrison T, Rowley DA, et al. IFN-gamma- and TNF-dependent bystander eradication of antigen-loss variants in established mouse cancers[J]. J Clin Invest, 2008, 118(4):1398-1404.
[60]苗明三.实验动物和动物实验技术[M].北京:中国中医药出版社, 1997.
[2] El-Serag HB, Rudolph KL. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis[J]. Gastroenterology, 2007, 132(7):2557-2576.
[3] Motola-Kuba D, Zamora-Valdés D, Uribe M, et al. Hepatocellular carcinoma. An overview[J]. Ann Hepatol, 2006, 5(1):16-24.
[4] Luo RH, Zhao ZX, Zhou XY, et al. Risk factors for primary liver carcinoma in Chinese population[J]. World J Gastroenterol, 2005, 11(28):4431-4434.
[5] Bosch FX, Ribes J, Díaz M, et al. Primary liver cancer: worldwide incidence and trends[J]. Gastroenterology, 2004, 127(5 Suppl 1):S5-S16.
[6] Marrero CR, Marrero JA. Viral hepatitis and hepatocellular carcinoma[J]. Arch Med Res, 2007, 38(6):612-620.
[7] Marrero JA. Hepatocellular carcinoma[J]. Curr Opin Gastroenterol, 2005, 21(3):308-312.
[8] Gomaa AI, Khan SA, Toledano MB, et al. Hepatocellular carcinoma: epidemiology, risk factors and pathogenesis[J]. World J Gastroenterol, 2008, 14(27):4300-4308.
[9]乔凤娟,梁睿,牛凤兰,等.东北菱对鼠肝癌H22细胞凋亡及细胞周期进程的影响[J].中国卫生工程学, 2006, 5(6):364-365.
[10]牛凤兰,董威严,乔凤娟.菱角提取物对荷瘤小鼠免疫功能的影响[J].食品科学, 2006, 27(7):233-235.
[11]吕喆,龚守良,牛凤兰,等.菱角纯化物三羟基苯甲酸二聚体对肝癌SMMC-7721细胞凋亡及细胞周期进程的影响[J].吉林大学学报(医学版), 2006, 32(5):788-790.
[12] Niu FL, Wang XD, Wang YL, et al. Trihydroxybenzoic acid dimer-induced apoptosis effect in vitro[J]. Chem Res Chinese U, 2005, 21(4):463-467.
[13]赵宏艳,张皖东,吕诚,等.灵芝提取物对H22荷瘤小鼠抗肿瘤作用的实验研究[J].中国中药杂志, 2008, 33(7):842-845.
[14]宁安红,曹婧,黄敏,等.灵芝多糖对小鼠肿瘤的抑制作用及免疫指标的观察[J].肿瘤研究与临床, 2004, 16(3):147-149.
[15]彭玉娜,谭获,王颖超,等.灵芝孢子粉对兔移植性肿瘤生存期及外周血的影响[J].时珍国医国药, 2008, 19(7):1737-1738.
[16]李颖博,李宇华,王瑞,等.灵芝多糖对肿瘤细胞与内皮细胞相互作用的影响[J].中国药理学通报, 2008, 24(2):250-253.
[17] Fujii E, Suzuki M, Matsubara K, et al. Establishment and characterization of in vivo human tumor models in the NOD/SCID/gamma(c)(null) mouse[J]. Pathol Int, 2008, 58(9):559-567.
[18] Kerbel RS. Human tumor xenografts as predictive preclinical models for anticancer drug activity in humans: better than commonly perceived-but they can be improved[J]. Cancer Biol Ther, 2003, 2(4 Suppl 1):S134-139.
[19] Uckun FM. Severe combined immunodeficient mouse models of human leukemia[J]. Blood, 1996, 88(4):1135-1146.
[20] Sausville EA, Burger AM. Contributions of human tumor xenografts to anticancer drug development[J]. Cancer Res, 2006, 66(7):3351-3354, discussion 3354.
[21] Teicher BA. Tumor models for efficacy determination[J]. Mol Cancer Ther, 2006, 5(10):2435-2443.
[22] Carver BS, Pandolfi PP. Mouse modeling in oncologic preclinical and translational research[J]. Clin Cancer Res, 2006, 12(18):5305-5311.
[23] Khanna C, Hunter K. Modeling metastasis in vivo[J]. Carcinogenesis, 2005, 26(3):513-523.
[24] Ohsugi T, Yamaguchi K, Kumasaka T, et al. Rapid tumor death model for evaluation of new therapeutic agents for adult T-cell leukemia[J]. Lab Invest, 2004, 84(2):263-266.
[25] Loisel S, Ster KL, Quintin-Roue I, et al. Establishment of a novel human B-CLL-like xenograft model in nude mouse[J]. Leuk Res, 2005, 29(11):1347-1352.
[26] Macor P, Secco E, Zorzet S, et al. An update on the xenograft and mouse models suitable for investigating new therapeutic compounds for the treatment of B-cell malignancies[J]. Curr Pharm Des, 2008, 14(21):2023-2039.
[27] Reuther T, Kübler AC, Staff CJ, et al. The RAG 2 mouse model for xenografted human oral squamous cell carcinoma[J]. Contemp Top Lab Anim Sci, 2002, 41(2):31-35.
[28] Fichtner I, Rolff J, Soong R, et al. Establishment of patient-derived non-small cell lung cancer xenografts as models for the identification of predictive biomarkers[J]. Clin Cancer Res, 2008, 14(20):6456-6468.
[29] Viaggi M, Dagrosa MA, Belli C, et al. A new animal model for human undifferentiated thyroid carcinoma[J]. Thyroid, 2003, 13(6):529-536.
[30] Wang Y, Sudilovsky D, Zhang B, et al. A human prostatic epithelial model of hormonal carcinogenesis[J]. Cancer Res, 2001, 61(16):6064-6072.
[31] Ke N, Zhou D, Chatterton JE, et al. A new inducible RNAi xenograft model for assessing the staged tumor response to mTOR silencing[J]. Exp Cell Res, 2006, 312(15):2726-2734.
[32] Elkas JC, Baldwin RL, Pegram M, et al. A human ovarian carcinoma murine xenograft model useful for preclinical trials[J]. Gynecol Oncol, 2002, 87(2):200-206.
[33] Orazine CI, Hincapie M, Hancock WS, et al. A proteomic analysis of the plasma glycoproteins of a MCF-7 mouse xenograft: a model system for the detection of tumor markers[J]. J Proteome Res, 2008, 7(4):1542-1554.
[34] Henriksson KC, Almgren MA, Thurlow R, et al. A fluorescent orthotopic mouse model for reliable measurement and genetic modulation of human neuroblastoma metastasis[J]. Clin Exp Metastasis, 2004, 21(6):563-570.
[35] De Velasco MA, Tanaka M, Anai S, et al. GFP image analysis in the mouse orthotopic bladder cancer model[J]. Oncol Rep, 2008, 20(3):543-547.
[36] Cullinane C, Dorow DS, Kansara M, et al. An in vivo tumor model exploiting metabolic response as a biomarker for targeted drug development[J]. Cancer Res, 2005, 65(21):9633-9636.
[37] Haldar M, Randall RL, Capecchi MR. Synovial sarcoma: from genetics to genetic-based animal modeling[J]. Clin Orthop Relat Res, 2008 Sep;466(9):2156-2167. Epub 2008 Jun 18.
[38] Stell A, Biserni A, Della Torre S, et al. Cancer modeling: modern imaging applications in the generation of novel animal model systems to study cancerprogression and therapy[J]. Int J Biochem Cell Biol, 2007, 39(7-8):1288-1296.
[39] Malik S, Balkwill FR. Tumour necrosis factor[J]. Br Med J (Clin Res Ed), 1988, 296(6631):1214.
[40] Spriggs D, Imamura K, Rodriguez C, et al. Induction of tumor necrosis factor expression and resistance in a human breast tumor cell line[J]. Proc Natl Acad Sci U S A, 1987, 84(18):6563-6566.
[41] Candolfi M, Jaita G, Pisera D, et al. Adenoviral vectors encoding tumor necrosis factor-alpha and FasL induce apoptosis of normal and tumoral anterior pituitary cells[J]. J Endocrinol, 2006, 189(3):681-690.
[42] Goodsell DS. The molecular perspective: tumor necrosis factor[J]. Oncologist, 2006, 11(1):83-84.
[43] Verhoef C, de Wilt JH, Grünhagen DJ, et al. Isolated limb perfusion with melphalan and TNF-alpha in the treatment of extremity sarcoma[J]. Curr Treat Options Oncol, 2007, 8(6):417-427.
[44] van Horssen R, Ten Hagen TL, Eggermont AM. TNF-alpha in cancer treatment: molecular insights, antitumor effects, and clinical utility[J]. Oncologist, 2006, 11(4):397-408.
[45] Bellone M, Mondino A, Corti A. Vascular targeting, chemotherapy and active immunotherapy: teaming up to attack cancer[J]. Trends Immunol, 2008, 29(5):235-241.
[46] Williams GM. Antitumor necrosis factor-alpha therapy and potential cancer inhibition[J]. Eur J Cancer Prev, 2008, 17(2):169-177.
[47] Suganuma M, Kuzuhara T, Yamaguchi K, et al. Carcinogenic role of tumor necrosis factor-alpha inducing protein of Helicobacter pylori in human stomach[J]. J Biochem Mol Biol, 2006, 39(1):1-8.
[48] Alexander HR Jr, Bartlett DL, Libutti SK. Current status of isolated hepatic perfusion with or without tumor necrosis factor for the treatment of unresectable cancers confined to liver[J]. Oncologist, 2000, 5(5):416-424.
[49] Bubeník J. Interleukin-2 therapy of cancer[J]. Folia Biol (Praha), 2004, 50(3-4):120-130.
[50] Rosenberg SA. The development of new immunotherapies for the treatment ofcancer using interleukin-2[J]. Ann Surg, 1988, 208(2):121-135.
[51] Uchiyama T. Interleukin-2 and its receptor in adult T cell leukemia[J]. Jpn J Med, 1991, 30(6):602-606
[52] Overwijk WW, Theoret MR, Restifo NP. The future of interleukin-2: enhancing therapeutic anticancer vaccines[J]. Cancer J Sci Am, 2000, 6 Suppl 1:S76-80.
[53] Atkins MB, Regan M, McDermott D. Update on the role of interleukin 2 and other cytokines in the treatment of patients with stage IV renal carcinoma[J]. Clin Cancer Res, 2004, 10(18 Pt 2):6342S-6346S.
[54] McDermott DF. Update on the application of interleukin-2 in the treatment of renal cell carcinoma[J]. Clin Cancer Res, 2007, 13(2 Pt 2):716s-720s.
[55] Adolf GR. Structure and effects of interferon-gamma[J]. Oncology, 1985, 42 Suppl 1:33-40.
[56] Blankenstein T, Qin Z. The role of IFN-gamma in tumor transplantation immunity and inhibition of chemical carcinogenesis[J]. Curr Opin Immunol, 2003, 15(2):148-154.
[57] Qin Z, Schwartzkopff J, Pradera F, et al. A critical requirement of interferon gamma-mediated angiostasis for tumor rejection by CD8+ T cells[J]. Cancer Res, 2003, 63(14):4095-4100.
[58] Merchant MS, Yang X, Melchionda F, et al. Interferon gamma enhances the effectiveness of tumor necrosis factor-related apoptosis-inducing ligand receptor agonists in a xenograft model of Ewing's sarcoma[J]. Cancer Res, 2004, 64(22):8349-8356.
[59] Zhang B, Karrison T, Rowley DA, et al. IFN-gamma- and TNF-dependent bystander eradication of antigen-loss variants in established mouse cancers[J]. J Clin Invest, 2008, 118(4):1398-1404.
[60]苗明三.实验动物和动物实验技术[M].北京:中国中医药出版社, 1997.