血管生成与DMBA诱发的大鼠乳腺癌转移的关系

目的：探讨肿瘤血管生成与乳腺癌转移的关系。方法：采用二甲基苯蒽（ＤＭＢＡ）诱发的大鼠乳腺癌模型,以免疫组织化学染色方法检测肿瘤组织微血管密度（ＭＶＤ）及血管内皮生长因子（ＶＥＧＦ）表达。结果：转移组原发瘤灶ＭＶＤ及瘤细胞ＶＥＧＦ表达显著的高于非转移组（分别Ｐ〈０．００１和Ｐ,０．０２）及良性肿瘤组（两者均Ｐ〈０．００１）。结论：ＶＥＧＦ介导的血管生民在促进乳腺癌转移的过程中可能起重要作用。     

乳腺癌肿瘤血管生成的临床病理学意义

肿瘤血管生成（ＴｕｍｏｒＡｎｇｉｏｇｅｎｅｓｉｓ，ＴＡ）是目前肿瘤研究的重要课题，其研究目的在于探讨ＴＡ的预后价值以及预测肿瘤对抗血管生成药物的反应。本文应用第八因子相关抗原多克隆抗体对６５例乳腺浸润性导管癌（以下简称乳腺癌）进行了微血管的定量研究。结果显示：腋下淋巴结阳性病例组织的微血管密度（ＭｉｃｒｏｖｅｓｓｅｌＤｅｎｓｉｔｙ，ＭＶＤ）（１２９．７±４４．９）明显高于腋下淋巴结阴性（ＮｏｄｅＮｅｇａｔｉｖｅＢｒｅａｓｔＣａｎｃｅｒ，ＮＮＢＣ）病例组的ＭＶＤ（７９．６±３３．６），差异呈极显著性（Ｐ＜０．００１）；发生术后复发及远处转移的病例的ＭＶＤ均值高达１４５．３；以上结果提示乳腺癌ＭＶＤ与肿瘤转移、复发均密切相关。我们认为乳腺癌ＭＶＤ可反映其血供状态，ＭＶＤ高的病例微血管丰富，肿瘤组织生长快，癌细胞易于进入微循环而发生转移     

Thrombospondin 1 Triggers Osteosarcoma Cell Metastasis and Tumor Angiogenesis

Osteosarcomas, especially those with metastatic or unresectable disease, have limited treatment options. The antitumor effects of pharmacologic inhibitors of angiogenesis in osteosarcomas are hampered in patients by the rapid development of tumor resistance, notably through increased invasiveness and accelerated metastasis. Here we demonstrated that thrombospondin 1 (TSP-1) is a potent inhibitor of the growth and metastasis of the osteosarcoma cell line MG-63. Moreover, we demonstrate that upregulation of TSP-1 facilitated expression of vasculostatin in MG-63 cells. In angiogenesis assays, overexpression of TSP-1 inhibited MG-63 cells and induced tube formation of human umbilical vein endothelial cells (HUVECs) in a CD36-dependent fashion. Finally, in xenografted tumors, we observed that TSP-1 overexpression inhibited angiogenesis and tumor growth. These results provided strong evidence for an important role of the TSP-1/CD36/vasculostatin signaling axis in mediating the antiangiogenic activity of osteosarcoma.     

乙酰肝素酶与大肠癌浸润、转移和血管生成的相关性

 目的 探讨乙酰肝素酶与大肠癌浸润、转移和血管生成之间的关系。方法 应用原位杂交方法检测乙酰肝素酶mRNA在95例大肠癌组织中的定位及表达。并用免疫组化方法对全部标本进行CDl05染色，记数肿瘤微血管密度（microvessel density，MVD），分析乙酰肝素酶mRNA表达与大肠癌浸润、转移和MVD之间的关系。结果 95例大肠癌组织中，乙酰肝素酶mRNA阳性表达49例（51．57％），MVD平均值为（72．1±20．6）；阴性表达46例（48．42％），MVD平均值为（41．3±12．4），差异有统计学意义（P〈0．01）。乙酰肝素酶tuRNA表达与大肠癌组织浸润深度、淋巴结转移及MVD有关（P〈0．05）。结论 乙酰肝素酶可促进大肠癌的浸润、转移和血管生成，可作为反映大肠癌生物学行为的客观指标。     

Correlation between Expression of P38 MAPK-Signaling and uPA in Breast Cancer

OBJECTIVE To study the expression of phosphorylated p38 mitogen-activated protein kinase(p-p38)and uPA and the correlation of their expression with breast cancer clinicopathological characteristics,and to investigate the role of the p38MAPK-signaling pathway in regulating uPA expression in breast cancer cells. METHODS Immunohistochemistry(S-P) was used to test the expression of p-p38 and uPA in 60 specimens of breast cancer tissues.Western blots were adopted to detect expression of the p-p38 and uPA proteins in MDA-MB-231 and MCF-7 breast cancer cells,and uPA expression a er treatment with SB203580,a specific inhibitor of p38 MAPK. RESULTS The positive rate of the p-p38 protein and uPA protein expression in the breast cancer tissues was 56.7% and 60.0%,respectively.The expression of p-p38 was positively related to the expression of uPA(r=0.316,P<0.05).The expression of p-p38 and uPA was related to lymph node metastas is and the TNM stage(P<0.05),but it was not related to the patient’s age or tumor size(P>0.05).The expression of p-p38 and uPA in MDA-MB-231 cells was higher than that in MCF-7 cells.SB203580 inhibited the p38 MAPK pathway and reduced uPA protein expression. CONCLUSION The p38 MAPK-signaling pathway promotes breast cancer malignant progression by up-regulating uPA expression,and it may be an important process in breast cancer invasion and metastasis.     

Farnesyl thiosalicylic acid inhibits the growth of melanoma cells through a combination of cytostatic and pro-apoptotic effects

Novel classes of drug that interfere with the signalling of the small G-protein Ras, the so-called Ras antagonists, are showing much promise as novel anti-cancer agents. In this study, we demonstrate that the novel Ras antagonist farnesylthiosalicylic acid (FTS) inhibits the growth of Colo 853 melanoma cells through a combination of cytostatic and pro-apoptotic effects. Furthermore, these phenomena are seen under conditions of cell attachment and in the presence of serum. Treatment of Colo 853 cells with FTS led to time-dependent inhibition of constitutive Akt, retinoblastoma protein (pRB) and ERK activity, with a concurrent loss of Akt expression. Inhibition of Akt and ERK activity induces apoptosis in other human cancer cell lines. Here it is demonstrated that inhibition of Akt, or ERK and Akt in combination, leads to cell cycle arrest but not apoptosis in melanoma cells. FTS treatment was also found to upregulate activity of the stress-activated p38 MAP kinase. Inhibition of p38 MAP kinase, using the selective inhibitor SB 203580, followed by FTS treatment, significantly increased the proportion of apoptotic cells after 72 hr, possibly suggesting a modulatory role for p38 MAP kinase in FTS-induced melanoma cell apoptosis.     

Efficacy of the Angiogenesis Inhibitor O-(chloroacetyl-carbamoyl)fumagillol (AGM-1470) on Osteosarcoma Growth and Lung Metastasis in Rats

The efficacy of the anti-angiogenic agent, O-(chloroacetyl-carbamoyl)fumagillol(AGM-1470), against primary tumor growth and spontaneous lungmetastasis was evaluated experimentally using a transplantableosteosarcoma line in rats previously established in our laboratory.Male Fischer 344 rats bearing the tumor with a high potentialfor metastasis received intermittent or continuous subcutaneousadministrations of AGM-1470. Both treatment regimens resultedin significant inhibitions of spontaneous lung metastasis andprimary tumor growth in a dose-dependent manner, with continuousadministration of AGM-1470 exerting the most pronounced inhibitoryeffects on both parameters     

Angiogenesis,Metastasis, and Endogenous Inhibition

Angiogenesis and the development of metastases are intrinsically connected. Experimental data suggest that establishment and growth of metastases are influenced by soluble factors secreted from the originating solid tumor. Among these factors are so-called endogenous inhibitors of angiogenesis which keep metastasis in a non-proliferating quiescent state. For a number of tumors it has been shown that this dormant state is mediated through inhibition of angiogenesis. This dormant state is characterized by normal proliferation, increased apoptosis, and insufficient neovascularization. Removal of inhibiting anti-angiogenic factors led to growth of dormant metastases. A number of endogenous inhibitors have been identified and have shown success in experimental therapeutic trials. This might be of special interest for the treatment of cerebral metastases which are the most common type of malignant brain tumors. Similar to the spread of metastases, it is known that single glioma cells can be found in distant parts of the brain. While local recurrence is a common phenomenon in glioma, formation of clinical apparent distant metastasis occurs rarely. Several lines of evidence suggest that growth inhibition of remote glioma cells may be mediated by an endogenous inhibitory mechanism.     

VEGF在大鼠脾内移植瘤模型中的表达及其与肿瘤血管形成、侵袭和转移的关系

目的探讨VEGF在大鼠移植瘤模型中的表达及意义。方法采用ELISA检测大鼠血清VEGF浓度,采用免疫组化法检测VEGF在肿瘤组织中的表达,F8因子肿瘤微血管染色分析肿瘤血管形成。结果实验大鼠血清VEGF水平接种后第一天明显升高,后两周保持稳定。大鼠脾内移植瘤模型中血清VEGF表达与肿瘤组织微血管密度、肝转移显著相关,γ=0.92,P〈0.05。肝转移肿瘤细胞胞质VEGF染色较深。16只大鼠移植瘤模型中肝转移9例,血清VEGF浓度为（2.57±0.68）ng/ml,未发生肝转移7例,VEGF浓度为（1.83±0.37）ng/ml（P〈0.05）;腹腔播散10例,VEGF浓度为（2.47±0.71）ng/ml,无腹腔播散6例,VEGF浓度为（1.86±0.39）ng/ml（P〈0.05）;出现血性腹腔积液11例,VEGF浓度为（2.47±0.66）ng/ml,未见血性腹腔积液5例,VEGF浓度为（1.75±0.38）ng/ml（P〈0.05）。结论实验大鼠中血清VEGF表达与肿瘤血管密度、肝转移、腹腔播散、血性腹腔积液有关。     

Inhibition of Liver Metastasis of Human Gastric Carcinoma by Angiogenesis Inhibitor TNP-470

The anti-tumor and anti-metastatic effects of TNP-470, an angiogenesis inhibitor, and mitomycin C (MMC), a representative anti-neoplastic agent, were investigated using our established liver-metastasizing gastric carcinoma line, AZ-H5c. AZ-H5c was injected into the spleen of nude mice which had been randomly divided into 4 groups; a control group given saline solution, a group receiving 15 mg/kg TNP-470, a group receiving 30 mg/kg TNP-470 and a group receiving 2 mg/kg MMC. TNP-470 was given s.c. on alternate days for 5 weeks from day 10 after intrasplenic injection, and MMC was administered intraperitoneally (i.p.) once a week from day 10 after intrasplenic injection. In the control group, liver metastasis developed in 13 of 16 mice (81%). Liver metastasis developed in 6 of 11 mice (55%) receiving MMC. In contrast, liver metastasis developed in 4 of 8 mice (50%) receiving 15 mg/Kg TNP-470, and in 0 of 14 mice (0%) receiving 30 mg/kg TNP-470. However, TNP-470 had no effect on the tumor growth. These results indicate that the angiogenesis inhibitor TNP-470 has a strong inhibitory activity against in vivo liver metastasis of human gastric carcinoma.     

大肠癌基质溶素与血管生成的关联性

 目的 探讨大肠癌基质溶素(Matrilysin，MMP-7)表达和肿瘤血管生成的关系。方法 用免疫组织化学法检测大肠癌中MMP-7表达及肿瘤微血管密度(Microvessel density．MVD)。结果 从正常肠黏膜、腺瘤、早期大肠癌到进展期大肠癌MVD表达呈上升趋势(P<0.05)。大肠癌MVD随肿瘤侵袭深度增加呈增高趋势(P<0.05)；有转移者高于无转移者(P<0.05)。50例大肠癌中，32例MMP-7阳性表达，MMP-7阳性表达组的MVD明显高于阴性组(P<0.05)；MMP-7和MVD显著正相关(r=0.7236，P<0.05)。结论 MMP-7可能通过促进肿瘤血管的形成参与大肠癌的发生、浸润和转移。     

塞来昔布联合奥沙利铂对人肺癌裸鼠移植瘤的治疗作用及其作用机制的研究

目的: 研究COX-2抑制剂与化疗药物奥沙利铂对人肺癌裸鼠移植瘤生长、Survivin表达和微血管生成的影响。 方法: 人肺癌A549细胞接种于裸鼠背部皮下,随机分为四组(对照组;奥沙利铂组;塞来昔布组;奥沙利铂和塞来昔布联合用药组)并给予相应的药物。测量肿瘤体积,42天后处死裸鼠,切取移植瘤组织检测COX-2,VEGF,Sur-vivin表达和微血管密度(MVD)。 结果: 塞来昔布组和奥沙利铂组抑瘤率分别为34.60%和46.70%,奥沙利铂和塞来昔布联合应用组抑瘤率为66.09%。免疫组织化学染色和RT-PCR结果分析显示:奥沙利铂组COX-2,VEGF表达显著高于对照组(P<0.05)。微血管密度与对照组比较无显著性差异(P>0.05)。奥沙利铂组Survivin表达低于对照组,塞来昔布组和联合用药组COX-2,VEGF表达和MVD低于对照组(P<0.05)。 结论: 塞来昔布可抑制人肺癌裸鼠移植瘤的生长、Survivin表达和血管生成。塞来昔布与奥沙利铂联合应用提高了奥沙利铂的抗肿瘤效果。     

Association of Vascular Endothelial Growth Factor Expression with Tumor Angiogenesis and with Early Relapse in Primary Breast Cancer

Angiogenesis is an independent prognostic indicator in breast cancer. In this report, the relationship between expression of vascular endothclial growth factor (VEGF; a selective mitogen for endothelial cells) and the microvessel density was examined in 103 primary breast cancers. The expression of VEGF was evaluated by immunocytochemical staining using anti-VEGF antibody. The microvessel density, which was determined by immunostaining for factor VIII antigen, in VEGF-rich tumors was clearly higher than that in VEGF-poor tumors ( P <0.01). There was a good correlation between VEGF expression and the increment of microvessel density. Furthermore, postoperative survey demonstrated that the relapse-free survival rate of VEGF-rich tumors was significantly worse than that of VEGF-poor tumors. It was suggested that the expression of VEGF is closely associated with the promotion of angiogenesis and with early relapse in primary breast cancer.     

An Inducible Melanoma Model Implicates a Role for RAS in Tumor Maintenance and Angiogenesis

The established tumor is maintained through complex and poorly understood host-tumor interactions guiding processes such as angiogenesis. The numerous and diverse genetic alterations that accompany tumor genesis raises questions as to whether experimental cancer-promoting mutations remain relevant to tumor maintenance. Utilizing a new doxycycline-inducible H-RAS^V12G INK4a null mouse melanoma model, we have shown that melanoma genesis and maintenance are strictly dependent upon H-RAS^V12G expression. Withdrawal of doxycycline and H-RAS^V12G down-regulation resulted in clinical and histological regression of primary and explanted tumors. Moreover, the initial stages of regression were highlighted by dramatic activation of apoptosis in the tumor cells as well as host-derived endothelial cells. These data provide genetic evidence that H-RAS^V12G plays a critical role in tumor maintenance and tumor angiogenesis     

Prevention of Hepatic and Peritoneal Metastases by the Angiogenesis Inhibitor FR-118487 after Removal of Growing Tumor in Mice

We established a mouse "primary tumor resection model" in which a transplanted tumor was resected after an orthotopic transplantation of colorectal cancer tissue to estimate the therapeutic effect of an angiogenesis inhibitor on metastasis. The angiogenesis inhibitor FR-118487 is a member of the fumagUlin family. Here, 1 mg/kg/day of FR-118487 was subcutaneously administered to nude mice for 1 week, 2 weeks, or 4 weeks through an osmotic pump. Liver metastasis developed in 7 of 9 control mice, 2 of 6 mice that underwent the tumor resection 2 weeks after transplantation (early resection), and in all 7 of the mice that underwent the tumor resection 4 weeks after transplantation (late resection). In the short treatment trial, the FR-118487 administration immediately after the early resection completely inhibited both hepatic and peritoneal metastases, whereas its administration after the late resection had no effect on liver metastasis. In the prolonged treatment trial, inhibitory effects of prolonged treatment with FR-118487 on both hepatic and peritoneal metastases after the late resection were clearly demonstrated. The mice of the resection-alone group all died within 106 days after tumor inoculation, due to metastases of colon carcinoma. In contrast, half of the mice that underwent resection and then received antiangiogenic therapy were alive at the end of the observation period (160 days after transplantation). In conclusion, the combination of surgery and subsequent antiangiogenic therapy may be useful to prevent the distant metastasis of colorectal cancer and to improve the prognosis of patients with colorectal cancer.     

Intracellular Localization and Biochemical Function of Variant β-Actin, which Inhibits Metastasis of B16 Melanoma

We analyzed the biochemical nature of βm-actin protein found in mouse B16 melanoma. When we carried out immunostaining with the antibody specific to βm-actin, filamentous immunofluorescence was observed in B16-F1, a low-metastatic cell line expressing βm-actin, but not in highly metastatic B16-F10 that did not express βm-actin. When a purified actin fraction containing βm-actin was polymerized and immunoprecipitated with anti-βm-actin antibody, the immunoprecipitate contained βm-, β- and γ-actin. This indicated that the βm-actin was incorporated into an actin filament together with β- and γ-actin in vitro , and this phenomenon was consistently suggested by cellular double immunostaining with anti-βm-actin and common anti-actin antibody. When the actin fraction containing βm-actin under a regular depolymerizing condition was subjected to immuno-adsorption assay using anti-βm antibody and protein-A Sepharose, the immunoadsorbed aggregates contained βm-, β-and γ-actin. This indicates that the actin fraction was not completely depolymerized and contained βm-actin-containing oligomers, which were too small to be precipitated with anti-βm-actin antibody alone. The incomplete depolymerization of the βm-actin-containing fraction was also suggested by the much lower DNase 1 inhibition activity of the βm-actin-containing fraction than that of β- and γ-actin fraction. Furthermore, a DNase 1 binding assay showed that cytoplasmic supernatant prepared from B16-F1 under a low-ionic condition contained less monomeric actin than the cytoplasmic preparation from B16-F10. These results suggested that βm-actin protein in B16 melanoma probably inhibits the dynamic conversion between the monomeric and polymerized forms of actin, leading to a decrease in cell motility and consequently the suppression of invasiveness and metastasis.     

金复康调控免疫衰老抑制肺癌转移

目的 探讨衰老对肺癌转移的影响,并揭示扶正中药金复康防治肺癌转移的作用机制。方法 不同月龄C57BL/6小鼠尾静脉注射Lewis肺癌细胞建立肺移植瘤模型;其中6月龄组小鼠采用金复康连续灌胃42天;观察小鼠肺部肿瘤转移情况;流式细胞术检测小鼠外周血中记忆T细胞、NK细胞比例的变化。结果 15月龄和6月龄小鼠较2月龄小鼠肺转移成瘤率显著上升（均P<0.05）。外周血记忆CD4+T细胞丰度与小鼠年龄呈正相关（2月龄 vs. 6月龄,P=0.041;2月龄 vs. 15月龄,P=0.041）。NK细胞丰度与年龄呈负相关（2月 vs. 6月,P=0.009;2月 vs. 15月,P=0.009）。金复康给药组小鼠的生存期较0.9%NaCl组有延长趋势,但差异无统计学意义,而给药组外周血NK细胞水平较0.9%NaCl组显著升高（P=0.029）。结论 衰老可促进肺癌转移发生,金复康延长小鼠生存期可能与延缓衰老和改善NK细胞水平有关。