共查询到20条相似文献,搜索用时 31 毫秒
1.
Jozef Bizik Diana Felnerova Marta Grofova Antti Vaheri 《Journal of cellular biochemistry》1996,62(1):113-122
Cultured human melanoma cells were found to secrete TGF-β mostly in latent biologically inactive form but in addition five of six melanoma cell lines studied produced in conditioned culture medium active TGF-β in the range from 370 to 610 pg per 106 cells per 24 h. A distinct characteristic of these melanoma cell lines is that they form active surface-bound plasmin by the activation of plasminogen with surface-bound tissue-type plasminogen activator. The present study was performed to assess the role of plasmin in the process of latent TGF-β activation in the melanoma cell lines. No direct correlation was found between cell-associated plasmin activity and the amount of active TGF-β present in the conditioned medium of individual cell lines. The melanoma cell lines exhibited diverse responses to exogenous active TGF-β1; three cell lines were growth-stimulated, two were growth-inhibited, and one had a very low sensitivity to the growth factor. The active TGF-β produced by the melanoma cells was found to inhibit the natural killer cell function of peripheral blood lymphocytes, suggesting that it may have an immunosuppressive effect and a role in the development of melanomas. © 1996 Wiley-Liss, Inc. 相似文献
2.
To investigate how the immune system responds to tumor self antigens, we used enhanced green fluorescent protein (EGFP) in B16 melanoma cells (B16-EGFP) and tested in the mouse lines expressing EGFP in thymic epithelial cells (3.1T-EGFP) or in antigen presenting cells (Get40), in comparison to the wild-type mouse. B16-EGFP cells were distinctively immunoedited in three mouse lines at the early phase, and the cells were completely eliminated only in the wild-type at the late phase, suggesting EGFP-specific tolerance is present in 3.1T-EGFP and Get40. The numbers of tumor-infiltrating T cells in all mouse lines were reversely correlated with the tumor sizes, suggesting dominant T cell mediated tumor elimination. When a soluble EGFP was immunized, surprisingly, the growth of B16-EGFP in Get40 mouse was promoted, while reduced in B6. Immunization did not make significant difference in the growth of tumors in 3.1T-EGFP. Detailed analyses showed the opposite directional changes in the numbers of B and CD8+T cells in B6 and Get40. In Get40 mice, the immunization significantly reduced the percentage of Gr1?CD11b+ cells, indicating that tolerance induction and breaking involve both adaptive and innate cells differentially. Therefore, the strategy for a cancer vaccine should be carefully considered on the types of antigen expressing cell. 相似文献
3.
Urokinase-type and tissue-type plasminogen activators are essential for in vitro invasion of human melanoma cells 总被引:4,自引:0,他引:4
A Meissauer M D Kramer M Hofmann L J Erkell E Jacob V Schirrmacher G Brunner 《Experimental cell research》1991,192(2):453-459
This study evaluates the contribution of two types of plasminogen activators (PAs; tissue-type PA (tPA) versus urokinase-type PA (uPA) toward the invasiveness of human melanoma cells in a novel in vitro assay. We identified two human melanoma cell lines, MelJuso and MeWo, expressing uPA or tPA as shown at mRNA, protein, and enzyme activity level. MelJuso cells produced uPA as well as plasminogen activator inhibitor-1 (PAI-1). The latter was, however, not sufficient to neutralize the cell-associated or secreted uPA activity. MeWo cells secreted tPA, but the enzyme was not found to be cell-associated. PAI-1 production by these cells was not detectable. Plasminogen activation and fibrinolytic capacity of both cell lines were reduced by anticatalytic monoclonal antibodies specific for the respective type of PA or by aprotinin. In a novel in vitro invasion assay, antibodies to PA as well as aprotinin decreased the invasiveness of both cell lines into a fibrin gel, Matrigel, or intact extracellular matrix. Our results confirm the importance of uPA-catalyzed plasminogen activation in tumor cell invasiveness. Furthermore, we provide evidence that tPA, beyond its key role in thrombolysis, can also be involved in in vitro invasion of human melanoma cells. 相似文献
4.
Mamoru Harada Koji Tamada Koichiro Abe Tieli Li Yasuhiro Onoe Hitoshi Tada Katsunori Tatsugami Takashi Ando Genki Kimura Kikuo Nomoto 《Cancer immunology, immunotherapy : CII》1998,47(4):198-204
A B16 melanoma-specific CD8+ T cell line (AB1) was established from the spleen cells of C57BL/6 mice cured of B16 melanoma with interleukin (IL)-12 treatment.
The AB1 line exclusively used T cell receptor Vβ11. The AB1 cells exhibited a cytolytic activity against both syngeneic B16 melanoma and allogeneic P815 mastocytoma, whereas
a cold inhibition assay revealed specificity of the AB1 cells against B16 melanoma. Their lostability to kill a class I loss
variant of B16 melanoma was restored by the transfection of H-2Kb gene. In addition, their interferon (IFN)-γ production was significantly suppressed by the addition of anti-H-2Kb monoclonal antibody, and RT-PCR analysis showed that the AB1 line expressed the mRNA encoding IFN-γ, but not IL-4 or IL-10.
The experiment using synthetic peptides of tyrosinase-related protein-2 (TRP-2) revealed that the AB1 cells could recognize
TRP-2181–188 peptide. Moreover, the AB1 cells showed an in vivo antitumor effect against established pulmonary metastases of B16 melanoma.
Overall, these results indicate that the Tc1-type Vβ11
+ AB1 cells exert an antitumor activity against syngeneic B16 melanoma through recognition of TRP-2181–188 peptide in an H-2Kb-restricted manner.
Received: 4 June 1998 / Accepted: 21 July 1998 相似文献
5.
Shingo Tamura Toshinori Bito Masamitsu Ichihashi Masato Ueda 《Pigment cell & melanoma research》2003,16(5):470-476
Genistein, a naturally occurring isoflavone found chiefly in soybeans, has been reported to be a potent antitumor agent. Genistein is presumed to exert multiple effects related to the inhibition of cancer growth. Metastatic melanoma is a chemotherapy‐refractory neoplasm. The present study was designed to explore the possible activity of genistein to inhibit the aberrant proliferation and to induce apoptosis of human malignant melanoma cells in cooperation with cisplatin treatment. Five human melanoma cell lines were utilized for these experiments. Genistein at physiologic concentrations (20 μM) did not induce apoptosis by itself but did enhance cisplatin‐induced apoptosis in all five human melanoma cell lines tested. The enhanced susceptibility among the cell lines was diverse. Changes in the expression of two anti‐apoptotic proteins, bcl‐2 and bcl‐xL, and one pro‐apoptotic protein, apoptotic protease activating factor‐1 (Apaf‐1), were examined. Genistein alone or cisplatin alone generally did not alter bcl‐2 expression or bcl‐xL expression, but slightly increased Apaf‐1 in some cell lines. The combined treatment with genistein and cisplatin significantly reduced bcl‐2 and bcl‐xL protein and increased Apaf‐1 protein expression. These data suggest that genistein therapy may enhance the chemosensitivity of melanoma patients. 相似文献
6.
D Santoli S F Radka M Igarashi B L Kreider S Ferrone 《Journal of immunology (Baltimore, Md. : 1950)》1986,137(2):400-407
Human activated T cells, long-term cultured in the presence of interleukin 2 (IL 2), were compared with autologous Epstein Barr virus-transformed B lymphoblastoid cell lines for expression of human leukocyte (HLA)-HLA-DR and -DQ antigens and for ability to induce proliferative responses in autologous and allogeneic lymphocytes. Immunofluorescence analysis performed with a panel of monoclonal antibodies (mAb) specific for HLA-DR or -DQ antigens did not reveal any significant difference in the expression of HLA-DR antigens but revealed reduced expression of HLA-DQ antigens on two out of four T cell lines tested. No obvious difference could be detected in the two-dimensional gel electrophoretic profile of HLA-DR and -DQ beta-chains synthesized by the autologous pairs of B and T cell lines. In contrast with previous reports, the IL 2-dependent cell lines consistently induced alloproliferative responses in standard 6-day mixed lymphocyte cultures; however, these responses were severalfold lower than those elicited by the autologous B lymphoid lines. Both anti-HLA-DR and anti-HLA-DQ mAb blocked the proliferative responses induced by the B cell lines but did not affect those generated by the T cell lines, suggesting that the latter cells induce T lymphocyte activation via a mechanism independent of HLA-DR or -DQ antigen expression on their surface. Addition of IL 2 to the mixed cultures with B cell lines as stimulators did not affect the outcome of the proliferative responses but partially or completely reversed the blocking activity of the mAb. In contrast, IL 2 significantly enhanced the alloproliferation induced by the T lymphoblastoid cell lines, and the anti-HLA class II mAb partially antagonized this effect. Taken together, these data suggest that unlike the HLA-DR and -DQ gene products on B cells, those on IL 2-dependent long-term cultured T cells do not play a direct or primary stimulatory role in the mixed lymphocyte reaction; the reduced levels of alloproliferation induced by the T cell lines are, at least in part, due to a defective production of endogenous IL 2 by the responder lymphocytes rather than to a defective expression of IL 2 receptors by the alloproliferative T cell subset; and the anti-HLA class II mAb in these cultures act only at the responder cell level, since they can efficiently block the enhancement of T cell proliferation triggered by exogenous IL 2, but not the proliferative responses induced by T cell lines in standard conditions. 相似文献
7.
8.
SARAH E. HILL JAYNE BUFFEY ANTHONY J. THODY IAN OLIVER STANLEY S. BLEEHEN SHEILA MAC NEIL 《Pigment cell & melanoma research》1989,2(3):161-166
In vitro melanocyte-stimulating hormone (MSH) stimulates melanogenesis in some, but not all, melanocytes and melanoma cells. In an attempt to explain this variation in response to αMSH, we examined cyclic adenosine monophosphate (cAMP) accumulation, tyrosinase activity, and melanin production in primary (1°) murine B16 melanoma cells and in two B16 cell lines (B16 F1 and B16 F10) that are known to respond to αMSH. In vivo all three B16 melanoma cell types produced pigmented tumours. In vitro αMSH increased tyrosinase activity and melanin content in the F1 and F10 cells but not in the B16 1° cells. αMSH, however, increased cAMP production in all three cell types, confirming that the inability of B16 1° cells to produce melanin in response to αMSH is not due to a lack of αMSH receptors or cAMP response to αMSH. Further, we present evidence for a separate pathway of melanogenesis that is independent of cAMP as calmodulin antagonists, which do not elevate cAMP, increased tyrosinase activity, and melanin production in both 1° and F1 cells. 相似文献
9.
M.J. Anasagasti A. Alvarez C. Avivi F. Vidal-Vanaclocha 《Journal of cellular physiology》1996,167(2):314-323
We have examined H2O2 production by in vitro enriched hepatic sinusoidal endothelium (HSE) during interleukin-1β (IL-1β) stimulation and B16 melanoma cell adhesion. Production of H2O2 was quantified by flow cytometry and multiwell plate-scanning fluorimetry of intracellular 2′, 7′-dichlorofluorescein (DCFH) oxidation in HSE. Under IL-1β treatment there was a 6-fold increase in endothelial cells producing H2O2 (67%) and a 4-fold augmentation in the Kupffer cell population (86%). The average H2O2 content per cell size unit significantly (P < 0.01) increased in endothelial cells (2.6-fold) and Kupffer cells (1.7-fold). In contrast to the homogeneity of Kupffer cells, H2O2 production intensity was largely heterogeneous in IL-1β-activated HSE. Enhancement of H2O2 production by IL-β-treated HSE started at the 4th h and peaked 2–3 h later. The addition of increasing concentrations of IL-1β to HSE for 4 h caused the progressive activation of H2O2 production by treated cells. The addition of 80 M excess of IL-1 receptor antagonist (IL-1Ra) 10 min before IL-1β treatment abrogated IL-1β-mediated enhancement of H2O2. From the 2nd h of B16 melanoma adhesion to HSE there was a significant (P < 0.05) enhancement of H2O2 content in HSE. This activation increased 2.25-fold by the 3rd h of coculture and had reduced again by the 5th h. IL-1Ra (80 ng/ml) given to HSE 10 min before melanoma cells abrogated the HSE response to melanoma cells. The addition of 1% paraformaldehyde (PFA)-fixed B16 melanoma cells to HSE did not affect H2O2 production response, indicating that HSE-activating agents were on the melanoma cell surface. Preincubation of B16 melanoma cells in the presence of 5 μg/ml anti-mouse IL-1β neutralizing antibody reduced the melanoma cell-induced HSE production of H2O2 by 80%. On the contrary, B16 melanoma cell-conditioned medium did not vary HSE production of H2O2 compared to control HSE. Western blot analysis of cytosolic and membrane sediments from B16 melanoma cells confirmed the presence of IL-1β (17.4 kDa) in both cell compartments. Thus, HSE responded to melanoma cell contact with a rapid production of H2O2. HSE activation was IL-1-dependent. This cytokine was directly provided to HSE by the cell surface of adhered melanoma cells. © 1996 Wiley-Liss, Inc. 相似文献
10.
Nikhil M. Vad Prabodh K. Kandala Sanjay K. Srivastava Majid Y. Moridani 《Chemico-biological interactions》2010,183(3):462-471
The aim of this study was to identify a phenolic prodrug compound that is minimally metabolized by rat liver microsomes, but yet could form quinone reactive intermediates in melanoma cells as a result of its bioactivation by tyrosinase. In current work, we investigated 24 phenolic compounds for their metabolism by tyrosinase, rat liver microsomes and their toxicity towards murine B16-F0 and human SK-MEL-28 melanoma cells. A linear correlation was found between toxicities of phenolic analogs towards SK-MEL-28 and B16-F0 melanoma cells, suggesting similar mechanisms of toxicity in both cell lines. 4-HEB was identified as the lead compound. 4-HEB (IC50 48 h, 75 μM) showed selective toxicity towards five melanocytic melanoma cell lines SK-MEL-28, SK-MEL-5, MeWo, B16-F0 and B16-F10, which express functional tyrosinase, compared to four non-melanoma cells lines SW-620, Saos-2, PC3 and BJ cells and two amelanotic SK-MEL-24, C32 cells, which do not express functional tyrosinase. 4-HEB caused significant intracellular GSH depletion, ROS formation, and showed significantly less toxicity to tyrosinase specific shRNA transfected SK-MEL-28 cells. Our findings suggest that presence of a phenolic group in 4-HEB is critical for its selective toxicity towards melanoma cells. 相似文献
11.
Begley J Vo DD Morris LF Bruhn KW Prins RM Mok S Koya RC Garban HJ Comin-Anduix B Craft N Ribas A 《Cancer immunology, immunotherapy : CII》2009,58(5):699-708
Several tumor immunotherapy approaches result in a low percentage of durable responses in selected cancers. We hypothesized
that the insensitivity of cancer cells to immunotherapy may be related to an anti-apoptotic cancer cell milieu, which could
be pharmacologically reverted through the inhibition of antiapoptotic Bcl-2 family proteins in cancer cells. ABT-737, a small
molecule inhibitor of the antiapoptotic proteins Bcl-2, Bcl-w and Bcl-xL, was tested for the ability to increase antitumor immune responses in two tumor immunotherapy animal models. The addition
of systemic therapy with ABT-737 to the immunization of BALB/c mice with tumor antigen peptide-pulsed dendritic cells (DC)
resulted in a significant delay in CT26 murine colon carcinoma tumor growth and improvement in survival. However, the addition
of ABT-737 to either a vaccine strategy involving priming with TRP-2 melanoma antigen peptide-pulsed DC and boosting with
recombinant Listeria monocytogenes expressing the same melanoma antigen, or the adoptive transfer of TCR transgenic cells, did not result in superior antitumor
activity against B16 murine melanoma. In vitro studies failed to demonstrate increased cytotoxic lytic activity when testing
the combination of ABT-737 with lymphokine activated killer (LAK) cells, or the death receptor agonists Fas, TRAIL-ligand
or TNF-alpha against the CT26 and B16 cell lines. In conclusion, the Bcl-2 inhibitor ABT-737 sensitized cancer cells to the
antitumor effect of antigen-specific immunotherapy in a vaccine model for the CT26 colon carcinoma in vivo but not in two
immunotherapy strategies against B16 melanoma. 相似文献
12.
S A Kudinov L A Kolesnik E M Makogonenko T A Shuliakovskaia 《Biokhimii?a (Moscow, Russia)》1989,54(11):1881-1887
The effect of heparin on the proteolytic and fibrinolytic activities of plasmin and plasminogen was studied. Heparin at a concentration of 6.3.10(-6) M did not change the caseinolytic activity of plasmin and plasminogen stimulated by streptokinase but suppressed their fibrinolytic activity. At concentrations from 2.10(-8) to 0.5.10(-6) M heparin increased, whereas at 1.10(-6)-4.10(-6) M reduced the time of desAAfibrin clot half-lysis by plasmin. Within the concentration range of 2.10(-8) to 4.10(-6) M heparin did not change the time of the clot half-lysis by glu-plasminogen and slightly decreased the time of fibrin clot half-lysis by lys-plasminogen in the presence of the tissue activator. It was supposed that heparin inhibits the fibrinolytic effect of plasmin by way of formation of complexes with plasmin and reduction of plasmin specificity to the solid phase substrate, i. e., polymeric fibrin. 相似文献
13.
Summary. The in vitro and in vivo effects of two flavonons, naringenin (NG) and hesperitin (HP) on the proliferation rate of highly metastatic murine B16-F10
melanoma cell were investigated. NG or HP treatment of melanoma cells produced a remarkable reduction of cell proliferation,
paralleled with both the lowering of the intracellular levels of polyamine, spermidine and spermine and the enhancement of
transglutaminase (TGase, EC 2.3.2.13) activity. Orally administered NG or HP in C57BL6/N mice inoculated with B16-F10 cells
affected the pulmonary invasion of melanoma cells in an in vivo metastatic assay. The number of lung metastases detected by a computerized image analyzer was reduced, compared to untreated
animals, by about 69% in NG-treated mice and by about 36% in HP-treated mice. Survival studies showed that 50% of the NG-treated
animals died 38 ± 3.1 days after tumor cell injection (control group: 18 ± 1.5 days) and HP-treated mice died 27 ± 2.3 days
after cell inoculation. Taken together, these findings provide further evidences for the potential anticancer properties of
dietary flavonoids as chemopreventive agents against malignant melanoma. 相似文献
14.
Teodoro Palomares Pedro Bilbao Ana Alonso-Varona Emilio Barberá-Guillem 《Cancer immunology, immunotherapy : CII》1995,40(5):292-298
The role of interleukin-2 (IL-2) on tumor growth of B16F10 melanoma cells was assessed in two sets of mice with different immune status: normal (immunocompetent) mice and immunodeficient mice. The two sets of animals were treated with cyclophosphamide (CY) or IL-2 alone or with a combined therapy of CY+IL-2. On days 6 and 10 after tumor cell injection, we evaluated the incidence of hepatic B16 melanoma metastases and the percentage of hepatic volume occupied by metastatic tissue. We observed that the CY alone (300 mg/kg, days 3 and 8 post-tumoral inoculation) significantly reduced tumor growth in all treated mice; however, CY proved more effective in normal recipients than in immunodeficient hosts. On the other hand, whereas administration of IL-2 alone (105 IU daily, from day 3 to day 7) in immunocompetent mice significantly reduced tumor growth on days 6 and 10, in immunodeficient mice, no significant differences were observed in tumor growth either on the 6th or on the 10th day, in comparison to control groups. Finally, when the combined CY+IL-2 therapy was administered, an antisynergistic effect between these therapeutic agents was achieved both in normal and in immunodeficient mice. Thus, the addition of low-dose IL-2 (25×103 IU daily, from day 4 to day 7) to high-dose CY (300 mg/kg, days 3 and 8) significantly increased tumor growth in both the early and later periods, compared to the effect of CY alone. It is concluded that exogenous IL-2 can facilitate tumor growth of B16 melanoma cells in vivo. 相似文献
15.
16.
T. Sakurai Muneo Yamada Seiichi Simamura Kazuo Motoyoshi 《Cancer immunology, immunotherapy : CII》1997,44(1):48-54
We studied the effect of recombinant human macrophage-colony-stimulating factor (rhM-CSF) on the formation of lung and liver
metastases following the i.v. injection of the B16 melanoma subline (B16 LiLu) into mice. When rhM-CSF was administered before
the B16 inoculation, the number of tumor metastases decreased in the lung and liver. However, the administration of rhM-CSF
after B16 inoculation did not produce an antimetastatic effect in the lung, but did in the liver. B16 cells labeled with 5-[125I]-iodo-2′-deoxyuridine (125I-dUrd) were injected and the arrest of tumor cell emboli was examined in the capillary beds of the lung and liver of mice
treated with either vehicle or rhM-CSF. In both groups, there were the same numbers of B16 cells in both the lung and the
liver 3 minutes after the B16 injection, and almost all tumor cells died within 24 h. However, the number of cells surviving
in the lung was decreased in mice injected with rhM-CSF (37%). There was no difference in the number of cells in the livers
of mice treated either with vehicle or rhM-CSF in the first 24 h after tumor cell injection. The administration of rhM-CSF
increased NK 1.1+ cells in the mouse spleen and facilitated NK activity in vivo. At the same time, the administration of an anti-NK 1.1 antibody
blocked the antimetastatic effect of rhM-CSF in the lung but not in the liver. The antibody was effective only when it was
injected before the B16 inoculation. These results suggest that the antimetastatic effect of rhM-CSF in the lung was mediated
by NK 1.1+ cells within 24 h of B16 injection. In contrast, the antimetastatic effect of rhM-CSF in the liver was mediated not only
by NK 1.1+ cells but also by other antimetastatic systems such as macrophages.
Received: 8 April 1996 / Accepted: 26 November 1996 相似文献
17.
S E Hill J Buffey A J Thody I Oliver S S Bleehen S Mac Neil 《Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society》1989,2(3):161-166
In vitro melanocyte-stimulating hormone (MSH) stimulates melanogenesis in some, but not all, melanocytes and melanoma cells. In an attempt to explain this variation in response to alpha MSH, we examined cyclic adenosine monophosphate (cAMP) accumulation, tyrosinase activity, and melanin production in primary (1 degree) murine B16 melanoma cells and in two B16 cell lines (B16 F1 and B16 F10) that are known to respond to alpha MSH. In vivo all three B16 melanoma cell types produced pigmented tumours. In vitro alpha MSH increased tyrosinase activity and melanin content in the F1 and F10 cells but not in the B16 1 degree cells. alpha MSH, however, increased cAMP production in all three cell types, confirming that the inability of B16 1 degree cells to produce melanin in response to alpha MSH is not due to a lack of alpha MSH receptors or cAMP response to alpha MSH. Further, we present evidence for a separate pathway of melanogenesis that is independent of cAMP as calmodulin antagonists, which do not elevate cAMP, increased tyrosinase activity, and melanin production in both 1 degree and F1 cells. 相似文献
18.
Chemically modified heparins as inhibitors of heparan sulfate specific endo-beta-glucuronidase (heparanase) of metastatic melanoma cells 总被引:2,自引:0,他引:2
To determine the significance of the heparan sulfate (HS) degradative endo-beta-glucuronidase (heparanase) in tumor invasion and metastasis and to develop possible antimetastatic agents, we synthesized specific inhibitors of this enzyme. We previously found that heparanase activity correlates with the lung colonization abilities of murine B16 melanoma cells and is inhibited by heparin [Nakajima, M., Irimura, T., Di Ferrante, N., & Nicolson, G. L. (1984) J. Biol. Chem. 259, 2283-2290]. In this study, heparin was chemically modified in order to determine which portions of its structure are responsible for heparanase inhibitory activity and to obtain heparanase inhibitors that have minimal additional biological effects, such as anticoagulation. N-Sulfate groups and O-sulfate in heparin were removed separately, and the resultant free amino groups were acetylated or resulfated. Heparin was also reduced at the carboxyl groups of uronic acid. The heparanase inhibitory activities of these heparin derivatives were examined by high-speed gel-permeation chromatography and by the use of radioactive HS immobilized on agarose beads. The results indicated that although N-sulfate and O-sulfate groups on glucosamine residues, and carboxyl groups on uronic acid residues, are important for heparanase inhibition, they are not essential for full activity. When highly metastatic B16-BL6 melanoma cells were incubated with N-acetylated N-desulfated heparin, N-resulfated N- and O-desulfated heparin, or carboxyl-reduced heparin and injected intravenously to syngenic C57BL/6 mice, significant reductions in the numbers of experimental melanoma lung metastases occurred. 相似文献
19.
Lapierre France; Holme Kevin; Lam Lun; Tressler Robert J.; Storm Neil; Wee Jennifer; Stack Robert J.; Castellot John; Tyrrell David J. 《Glycobiology》1996,6(3):355-366
Structural features of heparin potentially important for heparanase-inhibitoryactivity were examined by measuring the ability of heparin derivativesto affect the degradation of [3H]acetylated heparan sulphateby tumor cell heparanases. IC50 values were determined usingan assay which distinguished degraded from undegraded substrateby precipitation of the latter with cetylpyridinium chloride(CPC). Removal of heparin's 2-O-sulphate and 3-O-sul-phate groupsenhanced heparanase-inhibitory activity (50%). Removal of itscarboxyl groups slightly lowered the activity (18%), while combiningthe treatments abolished the activity. At least one negativecharge on the iduronic acid/idose moiety, therefore, is necessaryfor heparanase-inhibitory activity. Replacing heparin's N-sulphategroups with N-acetyl groups reduced its activity (37%). Comparingthis heparin derivative with 2,3-O-de-sulphated heparin, theplacement of sulphate groups appears important for activitysince the two structures have similar nominal linear chargedensity. In addition, unsubstituted uronic acids are nonessentialfor inhibition since their modification (periodate-oxidation/borohydride-reduction)enhanced rather than reduced heparanase-inhibitory activity.The most effective heparanase inhibitors (2,3-O-desulphatedheparin, and [periodate-oxidized, borohydride-reduced] heparin)were tested in the chick chorioallantoic membrane (CAM) bioassayfor anti-angiogenic activity and found to be at least as efficaciousas heparin. 2,3-O-desulphated heparin also significantly decreasedthe tumor growth of a subcutaneous human pancreatic (Ca-Pan-2)adenocarcinoma in nude mice and prolonged the survival timesof C57BL/6N mice in a B16-F10 melanoma experimental lung metastasisassay. angiogenesis chemically-modified heparins endoglycosidase hepara sulphate cancer 相似文献
20.
Genistein enhances the cisplatin-induced inhibition of cell growth and apoptosis in human malignant melanoma cells 总被引:3,自引:0,他引:3
Tamura S Bito T Ichihashi M Ueda M 《Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society》2003,16(5):470-476
Genistein, a naturally occurring isoflavone found chiefly in soybeans, has been reported to be a potent antitumor agent. Genistein is presumed to exert multiple effects related to the inhibition of cancer growth. Metastatic melanoma is a chemotherapy-refractory neoplasm. The present study was designed to explore the possible activity of genistein to inhibit the aberrant proliferation and to induce apoptosis of human malignant melanoma cells in cooperation with cisplatin treatment. Five human melanoma cell lines were utilized for these experiments. Genistein at physiologic concentrations (20 microM) did not induce apoptosis by itself but did enhance cisplatin-induced apoptosis in all five human melanoma cell lines tested. The enhanced susceptibility among the cell lines was diverse. Changes in the expression of two anti-apoptotic proteins, bcl-2 and bcl-xL, and one pro-apoptotic protein, apoptotic protease activating factor-1 (Apaf-1), were examined. Genistein alone or cisplatin alone generally did not alter bcl-2 expression or bcl-xL expression, but slightly increased Apaf-1 in some cell lines. The combined treatment with genistein and cisplatin significantly reduced bcl-2 and bcl-xL protein and increased Apaf-1 protein expression. These data suggest that genistein therapy may enhance the chemosensitivity of melanoma patients. 相似文献