75mGyX线全身照射与低剂量环磷酰胺合用对荷瘤小鼠肺肿瘤转移…

经低水平Ｘ线照射后，荷瘤小鼠肺肿瘤转移结节明显减少，与低剂量环磷酰胺（ＣＴＸ）合用，也可使荷瘤小鼠的肺转移结节减少，对荷瘤小鼠体内免疫功能的研究表明，小鼠脾脏细胞ＮＫ活性，白细胞介素Ⅱ（ＩＬ－２）诱导的ＬＡＫ活性ＬＴＲ及ＩＬ－２的反应性均显著增强，当与低剂量ＣＴＸ合用时上述抗肿瘤的免疫功能也提高，提示低剂量照射能通过有效地激活体内抗肿瘤免疫功能而具有显著的抗肿瘤转移效果，从而为探讨小剂量照射临床可     

X线全身照射与低剂量环磷酰胺合用对实验性肺转移的抑制作用

７５ｍＧｙ一次全身Ｘ线照射后，小鼠实验性肺肿瘤转移结节明显减少；与低剂量环磷酰胺（ＣＴＸ）合用，抗肿瘤转移作用亦较强。荷瘤小鼠７５ｍＧｙ照射后，脾脏ＮＫ细胞活性、ＬＡＫ细胞活性以及对白细胞介素Ⅱ（ＩＬ－２）的反应性明显增强或升高；与ＣＴＸ合用时上述抗肿瘤免疫功能也增强。结果表明：７５ｍＧｙＸ线全身照射可通过激活体内抗肿瘤免疫而具有有效的抗转移效果，它为探讨小剂量照射临床应用可能性提供了可靠的实验依据。     

75mGy X线全身照射与低剂量环磷酰胺合用对荷瘤小鼠肺肿瘤转移的治疗作用

经低水平Ｘ线照射后，荷瘤小鼠肺肿瘤转移结节明显减少；与低剂量环磷酰胺（ＣＴＸ）合用，也可使荷瘤小鼠的肺转移结节减少。对荷瘤小鼠体内免疫功能的研究表明，小鼠脾脏细胞ＮＫ活性、白细胞介素Ⅱ（ＩＬ－２）诱导的ＬＡＫ活性ＬＴＲ及时ＩＬ－２的反应性均显著增强，当与低剂量ＣＴＸ合用时上述抗肿瘤免疫功能也提高，提示低剂量照射能通过有效地激活体内抗肿瘤免疫功能而具有显著的抗肿瘤转移效果，从而为探讨小剂量照射临床应用可能性提供可靠的实验依据。     

75mGy X─线全身照射对荷瘤小鼠的抗肿瘤转移作用

经７５ｍＧｙＸ－线全身照射后，荷瘤小鼠肺肿瘤转移结节明显减少；与低剂量环磷酰胺（Ｃｙ）合用后，也可使荷瘤小鼠的肺转移结节减少。对荷瘤鼠体内免疫功能检测结果提示，小鼠脾脏ＮＫ活性、ＬＡＫ活性及对ＩＬ－２的反应性显著增强；当与低剂量Ｃｙ合用时，上述抗肿瘤免疫功能也提高，可见，低剂量照射能通过有效地激活体内抗肿瘤免疫功能而具有显著的抗肿瘤效果，它为低剂量照射在临床肿瘤治疗的应用提供了理论根据。     

小鼠75mGy X线全身照射对肿瘤细胞清除率的影响

７５ｍＧｙＸ线全身照射后，小鼠肝、脾等器官对肿瘤细胞清除率有明显提高。注入肿瘤细胞后２４ｈｒ肺、肝、脾中肿瘤细胞残留率照射组比对照组明显减少（Ｐ＜０．０１），肺部肿瘤转移结节也明显减少。有关免疫学指标检测结果表明，７５ｍＧｙＸ线全身照射后，机体巨噬细胞（Ｍ）吞噬作用、脾脏细胞ＮＫ活性和ＣｏｎＡ刺激的淋巴细胞掺入率（ＬＴＲ）明显提高或增强。实验结果提示，低剂量Ｘ线照射后小鼠机体对肿瘤细胞清除率的提高是通过激活体内抗肿瘤免疫功能而实现的。     

腺病毒介导的胞嘧啶脱氨酶自杀基因联合IL-2基因疗法的肿瘤治疗作用及免疫机理分析

目的以腺病毒作为载体，将大肠杆菌胞嘧啶脱氨酶（ＣＤ）基因与小鼠ＩＬ－２基因联合转移，研究其体内抗肿瘤作用及免疫机理。方法小鼠皮下接种黑色素瘤Ｂ１６Ｆ１０细胞后３天，肿瘤局部注射表达ＩＬ－２的重组腺病毒ＡｄＩＬ－２和表达ＣＤ的重组腺病毒ＡｄＣＤ，然后连续１０天给予５－氟胞嘧啶（５－Ｆｃ）３００ｍｇ／ｋｇ进行治疗。结果联合治疗组荷瘤小鼠皮下肿瘤结节的生长明显受到抑制，小鼠存活期明显长于ＡｄＩＬ－２、ＡｄＣＤ／５－Ｆｃ、ＡｄｌａｃＺ／５－Ｆｃ或ＰＢＳ组。经联合治疗后，小鼠脾细胞的ＮＫ活性和ＣＴＬ杀伤活性明显增强；肿瘤瘤体内ＣＤ４、ＣＤ８细胞浸润增加；肿瘤细胞表达Ｈ－２Ｋｂ和Ｂ７－１分子明显增加。结论联合应用自杀基因和ＩＬ－２基因治疗，一方面可以明显抑制荷瘤小鼠肿瘤生长，另一方面可以提高机体对肿瘤细胞免疫应答，增加机体的抗肿瘤作用，是肿瘤基因治疗中一条行之有效的途径。     

75mGyX一线全身照射对荷瘤小鼠的抗肿瘤转移作用

经７５ｍＧｙＸ一线全身照射后，荷瘤小鼠肺肿瘤转移结节明显减少；与低剂量环磷酰胺合用后，也可使荷瘤小鼠的肺转移结节减少。对荷瘤鼠体内免疫功能检测结果提示，小鼠脾脏ＮＫ活性、ＬＡＫ活性及对ＩＬ－２的反应性显著增强；当与低剂量Ｃｙ合用时，上述抗肿瘤免疫功能也提高，可见，低剂量照射能通过有效地激活体内抗肿瘤免疫功能而具有显著的抗肿瘤效果，它为低剂量照射在临床肿瘤治疗的应用提供了理论根据。     

甲氰咪呱对鼠体内IL—2／LAK抗肿瘤作用的影响

为探讨甲氰咪呱（ＣＩＭ）对鼠体内ＩＬ２／ＬＡＫ抗肿瘤作用的影响,将Ｂ１６细胞经尾静脉接种于Ｃ５７ＢＬ／６小鼠,第３ｄ分别应用ＤＨａｎｋｓ液、ＣＩＭ（１０ｍｇ／ｍｌ）＋ＣＩＭ（１μｇ／ｍｌ）诱导的脾细胞,ＩＬ２＋ＩＬ２（５００Ｕ／ｍｌ）诱导的ＬＡＫ细胞,ＣＩＭ（１０ｍｇ／ｍｌ）＋ＩＬ２＋ＣＩＭ（１μｇ／ｍｌ）和ＩＬ２（５００Ｕ／ｍｌ）共同诱导的ＬＡＫ细胞,观察鼠肺转移结节和生存期。结果显示,单用ＣＩＭ并不能减少肺转移结节和延长生存期;ＣＩＭ＋ＩＬ２／ＬＡＫ治疗组与ＩＬ２／ＬＡＫ治疗组相比,可减少肺转移结节（Ｐ＜０．０５）和显著延长生存期（Ｐ＜０．０１）。结果表明,ＣＩＭ能够增强ＩＬ２／ＬＡＫ抗肿瘤作用,可用于治疗肿瘤。     

蛋白激酶C抑制剂CJ9111抑制小鼠黑色素瘤B16细胞粘附和实验性转移

体外培养小鼠黑色素瘤Ｂ１６细胞，用不同剂量的ＣＪ９１１１分别作用不同时间，将处理后的细胞经尾静脉注入Ｃ５７ＢＬ／６小鼠体内，结果显示ＣＪ９１１１对Ｂ１６小鼠黑色素瘤细胞肺结节转移的抑制具有剂量和时间依赖性。药物作用２４小时对肺转移抑制作用的半数抑制浓度为８μｇ／ｍｌ（Ｐ＜０．０１），该值近似于ＣＪ９１１１对Ｂ１６细胞膜结合ＰＫＣ活力抑制作用的半数抑制浓度（ＩＣ５０６．３μｇ／ｍｌＰ＜０．０１）。药物作用后的Ｂ１６细胞与血管内皮细胞的粘附力下降，呈剂量依赖性。这表明ＰＫＣ可能是调节肿瘤细胞的转移重要因素，ＰＫＣ抑制剂有可能成为抗肿瘤转移的药物。     

AK细胞及环磷酰胺治疗肝癌的实验研究

为研究１ 种新的过继免疫化疗治疗肝癌的方法,采用肝癌细胞株Ｈ２２ 接种于近交系Ｂａｌｂ／ｃ 小鼠皮下,制成肿瘤模型。使用ＩＬ２ 、肿瘤活化的杀伤细胞（ＡＫ细胞）及环磷酰胺（Ｃｙ） 进行治疗,检测小鼠脾淋巴细胞ＮＫ、ＬＡＫ及ＣＴＬ 活性,用流式细胞仪检测Ｌ３Ｔ４ 亚群及Ｌｙｔ２ 亚群含量。结果表明在过继免疫化疗组小鼠ＬＡＫ 及ＣＴＬ活性明显高于其它治疗组（Ｐ＜ ０．０１）,荷瘤小鼠肿瘤结节生长较ＩＬ２ 组及Ｃｙ 组明显缓慢（Ｐ＜ ０．０１）,生存期也明显高于其它各治疗组（Ｐ＜０ ．０１）。该研究表明过继免疫化疗有较强的抗肿瘤作用。     

人参皂甙Rg3和核糖核酸酶抑制因子转基因对小鼠黑色素瘤的抑制作用研究

目的观察人参皂甙Rg3和核糖核酸酶抑制因子(RI)转基因对小鼠B16黑色素瘤肺转移的抑制作用和影响,探讨人参皂甙Rg3和RI抗肿瘤生长和转移作用的分子机制。方法制备转RI基因的B16黑色素瘤肺转移小鼠模型,对野生型对照组(W组)、空质粒转染组(B组)和RI转基因组(RI组)以及给予Rg3的野生型对照组(Rg3/W组)、空质粒转染组(Rg3/B组)和RI转基因组(Rg3/RI组)中,荷瘤小鼠肺重量、肿瘤转移灶数目、生存期和肿瘤组织微血管密度进行检测和分析。结果Rg3和RI转基因使荷瘤小鼠肺重量降低,肿瘤转移灶数目减少,其肺重量降低和肿瘤转移灶数目减少的程度以Rg3/RI组最明显,Rg3/B组、Rg3/W组和RI组次之,与W组和B组差异有显著性(P<0.01),Rg3和RI有一定的协同性。Rg3和RI可延长荷瘤小鼠的生存期,Rg3/RI组小鼠在观察期(1.5个月)内均存活,W组和B组小鼠全部死亡(至26d),且出现死亡的时间较早。经HE染色和第Ⅷ因子相关抗原的免疫组化分析显示,Rg3和RI使肺内瘤组织的微血管密度降低,降低的程度为Rg3/RI组>Rg3/B组>Rg3/W组>RI组>B组>W组。结论人参皂甙Rg3可增强RI转基因对小鼠黑色素瘤肺转移的抑制作用,人参皂甙Rg3和RI基因在抗肿瘤生长、转移及血管生成方面有协同作用。     

BALB／C小鼠黑色素瘤肿瘤模型的建立及对荷瘤鼠的影响

目的建立小鼠不同部位黑色素瘤模型，为不同研究目的提供合适的动物肿瘤模型。方法接种B16小鼠黑色素瘤细胞于BALB／C小鼠足部、腹腔和尾静脉，建立移植瘤模型和肺部转移瘤模型。观察B16实体瘤的形态学特点，接种细胞数与肿瘤生成的关系，肿瘤生长对小鼠生存时间、血清肿瘤坏死因子（sTNF）和唾液酸（SA）的影响。结果足部接种5X10瘤细胞后10天，肿瘤进入对数生长，成瘤率近100％；腹腔接种瘤细胞数大于5000个／只，成瘤率与接种细胞数成正比；接种3X10细胞／只，10天后在小鼠腹壁和肠系膜可见散在黑色素瘤结节，荷瘤鼠sTNF和SA水平明显高于正常小鼠，且与肿瘤的大小和数量成正比，小鼠平均存活59天；尾静脉接种后3周，肺部可见大小不等转移瘤灶，两肺平均瘤灶数可达192.11士92。结论B16黑色素瘤细胞株在BALB／C小白鼠具有很高的成瘤率和转移特性，且较C57／B16小黑鼠模型具有方便、稳定和观察容易等优点，是一较理想的模型。     

Comparison of the in vitro and in vivo effects of retinoids either alone or in combination with cisplatin and 5-fluorouracil on tumor development and metastasis of melanoma

Purpose  Retinoids have previously been reported to inhibit proliferation of melanoma cell lines in vitro. However, the relative antimetastatic efficacy of various retinoids on melanoma in vivo is unknown. Therefore, we investigated the effects of different retinoids on the invasion and metastasis of murine melanoma B16-F10 cells in vitro and in vivo. Based on the findings, the antitumor effects of a selected retinoid either alone or in combination with cisplatin were also investigated in a preclinical mouse melanoma model. Methods  Cell proliferation and invasion analyses of murine melanoma B16-F10 cells were assessed in the presence of different retinoids, either alone or in combination with cisplatin (CDDP) or 5-fluorouracil (5-FU). Experimental lung metastasis assay was performed in this study to investigate the antimetastatic efficacy of retinoids. Additionally, a mouse melanoma model was used to assess the antitumor efficacy of a selected retinoid in combination with cisplatin. Results  Retinoids showed significant antiproliferation and anti-invasion effects on murine melanoma B16-F10 cells. Pretreatment with retinoids increased the sensitivity to CDDP but not to 5-FU in in-vitro. Moreover, the number of metastatic colonies formed in the lungs of mice injected intravenously with B16-F10 cells was significantly reduced by injecting the respective retinoid once a day for 10 days. Treatment with a combination of cisplatin and 13-cis-retinoic acid resulted in a significant reduction in primary tumor size and the number of lung metastatic nodules in melanoma-bearing mice. Conclusion  These results suggest that retinoids not only exhibit antimetastatic effect, but also enhance the antitumor activity of cisplatin in vivo.     

Effects of non-motorized voluntary running on experimental and spontaneous metastasis in mice

The present study investigated the effects of non-motorized voluntary running on experimental metastasis of B16BL/6 melanoma and spontaneous metastasis of Lewis lung carcinoma (LLC) in male C57BL/6 mice. After 9 weeks of running, mice (n=30 per group) received an intravenous injection of B16BL/6 cells or a subcutaneous injection of LLC cells, and then they were continued with their running activities. Experiments were terminated 2 weeks after the intravenous injection of B16BL/6 cells or 2 weeks after surgical removal of the primary tumor from mice subcutaneously injected with LLC cells. Mice in the running group ran an average of 4-6 km/day for the duration of the experiment. Voluntary running reduced body weight compared with the sedentary controls, but there were no differences in the number and size of lung metastases between groups with either model. Voluntary running significantly reduced plasma insulin and leptin levels and increased adiponectin level in mice with and without LLC compared with the sedentary controls. Having LLC significantly increased plasma concentrations of vascular endothelial growth factor (VEGF), platelet-derived growth factor-BB (PDGF-BB), PDGF-AB and monocyte chemotactic protein-1 (MCP-1) in mice. Voluntary running significantly increased plasma PDGF-BB and PDGFAB, but not VEGF and MCP-1, in mice with LLC compared to their sedentary counterparts. In conclusion, non-motorized voluntary running was favorable to body weight and the expression of related adipokines, but at 4-6 km/day it did not affect either experimental or spontaneous metastasis in mice.     

Natural killer T cell ligand alpha-galactosylceramide inhibited lymph node metastasis of highly metastatic melanoma cells.

The role of natural killer T (NKT) cells in the prevention of multiple tumor metastasis was examined. The i.v. inoculation of a highly metastatic subline of B16-BL6 (B16-BL6-HM) melanoma cells resulted in the formation of metastatic nodules in lymph nodes in addition to lung, intrapleural cavity, and ovary. However, treatment of the mice with the NKT cell ligand alpha-galactosylceramide (alpha-GalCer) three times from 1 day after B16-BL6-HM melanoma inoculation caused a significant inhibition of multiple metastasis. Lymph node metastasis of B16-BL6-HM was almost completely blocked by alpha-GalCer treatment. This antimetastatic effect of alpha-GalCer was abolished in NKT cell-deficient mice. These results suggest that alpha-GalCer-activated NKT cells played a critical role in the prevention of lymph node metastasis of melanoma cells.     

Natural Killer T Cell Ligand α-Galactosylceramide Inhibited Lymph Node Metastasis of Highly Metastatic Melanoma Cells

The role of natural killer T (NKT) cells in the prevention of multiple tumor metastasis was examined. The i.v. inoculation of a highly metastatic subline of B16-BL6 (B16-BL6-HM) melanoma cells resulted in the formation of metastatic nodules in lymph nodes in addition to lung, intrapleural cavity, and ovary. However, treatment of the mice with the NKT cell ligand α-galactosylceramide (α-GalCer) three times from 1 day after B16-BL6-HM melanoma inoculation caused a significant inhibition of multiple metastasis. Lymph node metastasis of B16-BL6-HM was almost completely blocked by α-GalCer treatment. This antimetastatic effect of α-GalCer was abolished in NKT celldeficient mice. These results suggest that α-GalCer-activated NKT cells played a critical role in the prevention of lymph node metastasis of melanoma cells.     

Combination therapy with pions and schizophyllan (spg) for murine B-16 melanoma

The B-16 melanoma transplanted into C57BL/6 mice was used to investigate the antitumor effect of Schizophyllan (SPG) when applied alone and in combination with local irradiation using pions with 4 dose fractions of 2 Gy or 6 Gy each. SPG was given intramuscularly in a daily dose of 10 mg/kg body weight for 25 consecutive days from day 4 after the initiation of irradiation, and thereafter three times a week up to day 45. The antitumor effect was evaluated by the changes in tumor volume. survival, the size of metastatic lymph nodes grossly involved, and the number of pulmonary metastatic nodules on the surface of the lungs. After 24 Gy, significant differences were found between the group treated with combined pions and SPG, and the group treated with pions alone in terms of tumor volume change. survival and lymph node, and pulmonary metastases. However, when SPG was applied to non-irradiated tumors or to tumors irradiated with only 8 Gy, it had neither antitumor nor life-prolonging effect. From the present study, it seems that a SPG, as a Biological Response Modifier, has some adjuvant effect only where a limited number of tumor cells remain following pion irradiation. Combination therapy using SPG may, therefore, be advantageous for patients with complete response or good partial response to pion irradiation.     

Broad-spectrum anti-tumor and anti-metastatic DNA vaccine based on p62-encoding vector

Autophagy plays an important role in neoplastic transformation of cells and in resistance of cancer cells to radio- and chemotherapy. p62 (SQSTM1) is a key component of autophagic machinery which is also involved in signal transduction. Although recent empirical observations demonstrated that p62 is overexpressed in variety of human tumors, a mechanism of p62 overexpression is not known. Here we report that the transformation of normal human mammary epithelial cells with diverse oncogenes (RAS, PIK3CA and Her2) causes marked accumulation of p62. Based on this result, we hypothesized that p62 may be a feasible candidate to be an anti-cancer DNA vaccine. Here we performed a preclinical study of a novel DNA vaccine encoding p62. Intramuscularly administered p62-encoding plasmid induced anti-p62 antibodies and exhibited strong antitumor activity in four models of allogeneic mouse tumors – B16 melanoma, Lewis lung carcinoma (LLC), S37 sarcoma, and Ca755 breast carcinoma. In mice challenged with Ca755 cells, p62 treatment had dual effect: inhibited tumor growth in some mice and prolonged life in those mice which developed tumor size similar to control. P62-encoding plasmid has demonstrated its potency both as a preventive and therapeutic vaccine. Importantly, p62 vaccination drastically suppressed metastasis formation: in B16 melanoma where tumor cells where injected intravenously, and in LLC and S37 sarcoma with spontaneous metastasis. Overall, we conclude that a p62-encoding vector(s) constitute(s) a novel, effective broad-spectrum antitumor and anti-metastatic vaccine feasible for further development and clinical trials.     

A simple and effective method for cancer immunotherapy by inactivated allogeneic leukocytes infusion

Allogeneic mixed leukocytes reaction has been reported to activate vast numbers of T lymphocytes and produce large amounts of type 1 cytokines that are linked to an initiation of antitumor immunity. Using poor immunogeneic B16‐F10 and Lewis lung carcinoma (LLC) tumor model, we evaluated the effects of inactivated allogeneic leukocytes infusion (ALI) on the generation of antitumor immune response, as well as its effect on the primary and metastatic tumor. Allogeneic response promoted the generation of both specific and nonspecific antitumor immunity in an in vitro mixed lymphocytes‐tumor cell culture system. Introveinous infusion of mitotically inactivated allogeneic leukocytes resulted in increased type‐1 cytokines (including IL‐2 and IFN‐γ) release, proliferation of various lymphocyte subsets, and generation of both specific and nonspecific antitumor immune response. As a result of such immune response, ALI caused a delayed tumor growth and a prolonged survival in established B16‐F10 melanoma model. In LLC pulmonary spontaneous metastases model, ALI treatment significantly reduced postoperative tumor metastasis as the lung weights were far smaller than control group (0.16 vs. 0.34 g). Furthermore, after primary tumor resection and ALI treatment, 62.5% mice obtained long‐term survival (>120 days) and there were no tumor growths in these mice when they were rechallenged with the same tumor. These experiments demonstrate that inactivated ALI could activate innate and adoptive antitumor immune response. It would be a simple, effective and secured method for cancer immunotherapy. © 2008 Wiley‐Liss, Inc.     

Suramin augments the antitumor and antimetastatic activity of pentoxifylline in B16F10 melanoma

Rapid tumor growth and metastasis are 2 major problems associated with treatment of malignant melanoma. Therefore, drugs that can intervene these processes are of clinical importance. Pentoxifylline (PTX), a methyl xanthine derivative, has been shown to inhibit B16F10 melanoma tumor growth and metastasis. We hypothesized that suramin when combined with PTX enhances its antineoplastic effects, which we have examined using the B16F10 mouse melanoma model. Suramin in simultaneous or sequential combination potentiated the cytotoxic effects of PTX on B16F10 cells. PTX arrested cells in the G0-G1 phase and suramin augmented the effects. Both the drugs inhibited F10 adhesion to laminin, matrigel and collagen type IV and showed enhanced inhibition in combination The combination also demonstrated significantly higher inhibition in cell motility (p = 0.002) and invasion through matrigel (p = 0.005) as compared to the single agents. Suramin synergized with PTX in its effects on secretion of MMP-9 gelatinase. DBA2/J mice implanted with intradermal B16F10 tumor were used as a model to study tumor growth. Animals were intratumorally treated with 50 mg/kg of PTX, 10 mg/kg of suramin and their combinations. Simultaneous administration of the drugs inhibited tumor growth by 5- to 6-folds. Tumor growth was completely blocked in sequential regimen with regression in some cases. The number and size of metastatic nodules on lung was also reduced significantly by the combination treatment. In conclusion, the novel combination of PTX and suramin has synergistic antitumor and antimetastatic activity in B16F10 melanoma and may be a promising approach in treatment of patients suffering from malignant melanoma.