Solid Tumor Growth In Mice Research Articles

Hsa_circ_0069094 positively regulates the expression of oncogenic ZNF217 by competitively targeting miR-758-3p to promote the development of breast cancer.

To investigate the functions and potential mechanisms of hsa_circ_0069094 in this cancer. The expression of hsa_circ_0069094, zinc finger protein 217 (ZNF217) and microRNA-758-3p (miR-758-3p) was detected by quantitative polymerase chain reaction (qPCR), and the protein level of ZNF217 was detected by western blot. Cell proliferation was assessed using cell counting kit-8 (CCK-8) assay and colony formation assay. Cell cycle progression and cell apoptosis were determined using flow cytometry assay. Cell invasion and cell migration were monitored using transwell assay and wound healing assay. The protein levels of apoptosis-related proteins were quantified by western blot. The putative relationship between miR-758-3p and hsa_circ_0069094 and ZNF217 was confirmed using dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Xenograft model was constructed in mice to explore the role of hsa_circ_0069094 on solid tumor growth.Hsa_circ_0069094 and ZNF217 were highly expressed, while miR-758-3p was poorly expressed in tissues and cells of breast cancer. Hsa_circ_0069094 knockdown or ZNF217 knockdown inhibited cell proliferation, invasion and migration and induced cell apoptosis and cell cycle arrest in breast cancer cells. The inhibitory effects of hsa_circ_0069094 knockdown on cell malignant behaviors were abolished by ZNF217 overexpression. Hsa_circ_0069094 competed with ZNF217 for the binding site of miR-758-3p, and hsa_circ_0069094 positively regulated ZNF217 expression by competitively binding to miR-758-3p. Hsa_circ_0069094 knockdown also blocked solid tumor growth in mice. Collectively, Hsa_circ_0069094 played oncogenic effects in breast cancer by activating the expression of ZNF217 via competitively binding to miR-758-3p, which might be a novel strategy for breast cancer suppression.

Intratumoral nanobody-IL-2 fusions that bind the tumor extracellular matrix suppress solid tumor growth in mice.

Confining cytokine exposure to the tumors would greatly enhance cancer immunotherapy safety and efficacy. Immunocytokines, cytokines fused to tumor-targeting antibodies, have been developed with this intention, but without significant clinical success to date. A critical limitation is uptake by receptor-expressing cells in the blood, that decreases the dose at the tumor and engenders toxicity. Small-format immunocytokines, constructed with antibody fragments, are hypothesized to improve tumor specificity due to rapid systemic clearance. However, effective design criteria for small-format immunocytokines need further examination. Here, we engineer small interleukin-2 (IL-2) immunocytokines fused to nanobodies with nanomolar to picomolar affinities for the tumor-specific EIIIB domain of fibronectin (also known as EDB). Upon intravenous delivery into immunocompetent mice, such immunocytokines led to similar tumor growth delay as size-matched untargeted IL-2. Intratumoral (i.t.) delivery imparted improved survival dependent on affinity to EIIIB. I.t. administration offers a promising avenue to deliver small-format immunocytokines, given effective affinity for the tumor microenvironment.

Efficacy of ginsenoside Rg3 nanoparticles against Ehrlich solid tumor growth in mice.

Solid tumors are fairly common and face many clinical difficulties since they are hardly surgically resectable and broadly do not respond to radiation and chemotherapy. The current study aimed to fabricate ginsenoside Rg3 nanoparticles (Rg3-NPs) and evaluate their antitumor effect against Ehrlich solid tumors (EST) in mice. Rg3-NPs were fabricated using whey protein isolates (WPI), maltodextrin (MD), and gum Arabic (GA). EST was developed by the injection of mice with Ehrlich ascites cells (2.5 × 106). The mice were divided into a control group, EST group, and the EST groups that were treated orally 2weeks for with normal Rg3 (3mg/kg b.w.), Rg3-NPs at a low dose (3mg/kg b.w.), and Rg3-NPs at a high dose (6mg/kg b.w.). Serum and solid tumors were collected for different assays. The results revealed that synthesized Rg3-NPs showed a spherical shape with an average particle size of 20nm and zeta potential of -5.58mV. The in vivo study revealed that EST mice showed a significant increase in AFP, Casp3, TNF-α, MMP-9, VEGF, MDA, and DNA damage accompanied by a significant decrease in SOD and GPx. Treatment with Rg3 or Rg3-NPs decreased the tumor weight and size and induced a significant improvement in all the biochemical parameters. Rg3-NPs were more effective than Rg3, and the improvement was dose-dependent. It could be concluded that fabrication of Rg3-NPs enhanced the protective effect against EST development which may be due to the synergistic effect of Rg3 and MD, GA, and WPI.

Antioxidant activities of α-lipoic acid free and nano-capsule inhibit the growth of Ehrlich carcinoma.

To evaluate the relationship between oxidative stress, DNA damage, and suppression of tumor growth induced by lipoic acid free and nano-capsule. Lipoic acid nano-capsule was assessed by transmission electron microscopy (TEM). The antitumor activity of lipoic acid free and nano-capsule was investigated in Ehrlich solid tumor bearing mice. The endpoints measured were superoxide dismutase (SOD) enzyme activity, malondialdehyde (MDA) levels, DNA damage (comet assay), histopathological examination of tissues and tumor growth volume. Treatment with α-lipoic acid (A-LA) free and nano-capsule (1.2, 2.4mg/kg) showed a significant depression (67-28%) in MDA levels and elevated (38-98%) in SOD activity. Additionally, A-LA free and nano-capsule caused a 3-10-fold increase in comet parameters such as % tail DNA and suppressed Ehrlich solid tumor growth in mice. In conclusion, the present study revealed that A-LA free and nano-capsule have antitumor activity. This might be possible via its redox activity.

Polysaccharides from Chinese Herbal Lycium barbarum Induced Systemic and Local Immune Responses in H22 Tumor-Bearing Mice.

Lycium barbarum polysaccharide (LBP) is isolated from the fruit of Chinese herbal Lycium barbarum. Previous studies had demonstrated that LBP could inhibit tumor growth and enhance the immunity in mice. However, the effect of LBP on systemic and local immune responses in vivo, especially on phenotypic and functional changes of T cells, is still largely unknown. In the present study, we investigated the effects of LBP on systemic and local T cell-dependent antitumor immune responses in H22 tumor-bearing mice. The results showed that LBP could inhibit the solid tumor growth in mice, but showed little effect on the body weight or spleen index. Furthermore, LBP could maintain high levels of T cells in peripheral blood (PB), tumor draining lymph node (TDLN), and tumor tissue, prevent the increase of Tregs while promote infiltration of CD8+ T cells in tumor tissue, inhibit the production of TGF-β1 and IL-10 in serum, decrease the exhaustion phenotype of T cells, and maintain cytotoxicity of lymphocytes. Taken together, our results demonstrated that LBP simultaneously induced systemic and local immune responses in H22 tumor-bearing mice by alleviating immunosuppression and maintaining antitumor immune responses in mice.

Nickel(II) diacetyl monoxime-2-pyridyl hydrazone complex can inhibit Ehrlich solid tumor growth in mice: A potential new antitumor drug

The chief chemotherapeutic drug, cisplatin had common bad effects such as nephrotoxicity, ototoxicity and bone marrow depression. This led us to develop a new potential anticancer drug based on nickel metal ion that may be less toxic. Nickel(II) diacetyl monoxime-2-pyridyl hydrazone complex cytoprotective effect, superoxide dismutase (SOD)-like activity and anticancer activities were studied. In vitro, the complex showed SOD-like activity of 86.62%. It was capable to kill 90.2% of Ehrlich ascites carcinoma (EAC) cells and to protect 92.48% of human RBCs. In vivo, the complex lowered the tumor burden markedly in a concentration-dependent manner. Noticeably, solid tumor growth was suppressed; tumor volume and weight were reduced and mice life span was lengthened. The hematological indices were improved, catalase activity was re-elevated and malondialdehyde (MDA) level was reversed towards normal. Nucleic acids, cholesterol, triglycerides, liver enzymes, urea and creatinine contents were reduced to near normal ranges. Glutathione (GSH), SOD, albumin and total protein levels were increased. In conclusion, our results revealed that the complex has the ability to suppress Ehrlich solid tumor growth in mice with minimal side effects. This may possibly via its redox activity. Surprisingly, nickel complex antitumor activities were more potent than those of cisplatin.

Neem leaf glycoprotein prophylaxis transduces immune dependent stop signal for tumor angiogenic switch within tumor microenvironment.

We have reported that prophylactic as well as therapeutic administration of neem leaf glycoprotein (NLGP) induces significant restriction of solid tumor growth in mice. Here, we investigate whether the effect of such pretreatment (25µg/mice; weekly, 4 times) benefits regulation of tumor angiogenesis, an obligate factor for tumor progression. We show that NLGP pretreatment results in vascular normalization in melanoma and carcinoma bearing mice along with downregulation of CD31, VEGF and VEGFR2. NLGP pretreatment facilitates profound infiltration of CD8+ T cells within tumor parenchyma, which subsequently regulates VEGF-VEGFR2 signaling in CD31+ vascular endothelial cells to prevent aberrant neovascularization. Pericyte stabilization, VEGF dependent inhibition of VEC proliferation and subsequent vascular normalization are also experienced. Studies in immune compromised mice confirmed that these vascular and intratumoral changes in angiogenic profile are dependent upon active adoptive immunity particularly those mediated by CD8+ T cells. Accumulated evidences suggest that NLGP regulated immunomodulation is active in tumor growth restriction and normalization of tumor angiogenesis as well, thereby, signifying its clinical translation.

Subcutaneous administration of ketoprofen delays Ehrlich solid tumor growth in mice

Ketoprofen, a nonsteroidal anti-inflammatory drug (NSAID) has proven to exert anti-inflammatory, anti-proliferative and anti-angiogenic activities in both neoplastic and non-neoplastic conditions. We investigated the effects of this compound on tumor development in Swiss mice previously inoculated with Ehrlich tumor cells. To carry out this study the solid tumor was obtained from cells of the ascites fluid of Ehrlich tumor re-suspended in physiological saline to give 2.5x106cells in 0.05mL. After tumor inoculation, the animals were separated into two groups (n = 10). The animals treated with ketoprofen 0.1µg/100µL/animal were injected intraperitoneally at intervals of 24h for 10 consecutive days. Animals from the control group received saline. At the end of the experiment the mice were killed and the tumor removed. We analyzed tumor growth, histomorphological and immunohistochemical characteristics for CDC47 (cellular proliferation marker) and for CD31 (blood vessel marker). Animals treated with the ketoprofen 0.1µg/100µL/animal showed lower tumor growth. The treatment did not significantly influence the size of the areas of cancer, inflammation, necrosis and hemorrhage. Moreover, lower rates of tumor cell proliferation were observed in animals treated with ketoprofen compared with the untreated control group. The participation of ketoprofen in controlling tumor malignant cell proliferation would open prospects for its use in clinical and antineoplasic therapy.

Abstract B32: Cell penetrating Myosin-Va fragment covering the binding motif of DLC2: Evaluation of pro-apoptotic properties

Abstract Previous studies proposed that myosin-Va regulates apoptosis by sequestering pro-apoptotic Bmf to the actin cytoskeleton through dynein light chain-2 (DLC2). Adhesion loss or other cytoskeletal perturbations would unleash Bmf, allowing it to bind and inhibit pro-survival Bcl2 proteins. Recently, our research group has demonstrated that overexpression of a myosin-Va medial tail fragment (MVaf) harboring the binding site for DLC2 dramatically decreased melanoma cell viability. Morphological and molecular changes, including surface blebbing, mitochondrial outer membrane permeabilization, cytochrome-c and Smac release, as well as caspase-9/-3 activation and DNA fragmentation indicated that melanoma cells died of apoptosis. Besides, MVaf expression attenuated B16-F10 solid tumor growth in mice suggesting that this peptide may be useful as an apoptosis-inducing tool for basic cancer research. We suggest that MVaf induces apoptosis by sequestering DLC2 and DLC1, thereby unleashing the pair of sensitizer and activator BH3-only proteins Bmf and Bim. In view of such findings, we have proposed to develop a recombinant Tat-fusion MVaf to be explored in peptide-based therapeutic approaches. The native MVaf sequence was altered through the N-terminal addition of a transducible Tat-HA tag. EGFP was alternatively introduced into recombinant products to assess their subcellular localization. Bacterially expressed His-tagged fusion proteins were purified by immobilized metal chelate affinity chromatography on Ni-NTA beads, and transduction assays were initially performed in HeLa cells. The kinetics of Tat-HA-MVaf (THM, 13.7 kDa) and Tat-HA-EGFP-MVaf (THEM, 41.9 kDa) uptake, at concentrations ranging from 1 to 2 μM, was evaluated by confocal laser scanning microscopy (CLSM). Tat-HA-EGFP (THE, 33.9 kDa) was used as a control. Tat-HA-MVaf displayed a diffuse distribution throughout the cells in the first 5 min incubation, with a clear perinuclear accumulation after 1h incubation. THEM and THE fluorescence were detected in many developing vesicles adjacent to cell membrane, and distributed throughout the cytoplasm and nucleus after 5 min incubation, but became markedly weaker after 1h, suggesting proteolytic loss. Despite intense diffusion and intracellular accumulation of recombinant proteins, no significant changes in morphology and cell viability were detected by means of flow cytometry and MTT assay. Pull-down assay revealed that THEM but not THE binds to immobilized GST-DLC2 and to GST-DLC1 in a lesser extent, which confirms peptide-binding capacity to DLCs. On the other hand, pretreatment of cells with chloroquine, an inhibitor of endosomal acidification, sensitized cells to peptide treatment. We believe that Tat-mediated subcellular localization may have predominated, resulting in mislocalization of the peptide cargo. Also, one major drawback in using pTAT-conjugated peptides is their entrapment in cytoplasmic compartments and nucleus, resulting in limited availability and biological activity. One possible strategy is to produce recombinant peptides conjugated reversibly to Tat by a disulfide bond, which are disrupted upon intracellular delivery. Funding support. FAPESP / CNPq. Citation Format: Denise P. S. Leitão Mazzi, Cleidson P. Alves, Enilza Maria Espreafico. Cell penetrating Myosin-Va fragment covering the binding motif of DLC2: Evaluation of pro-apoptotic properties. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr B32.

Differential immune regulation of mir-155 in tumor growth (P2113)

Abstract MiR-155 is upregulated in both myeloid and lymphoid cells following their activation. To address the role of host miR-155 in tumor growth, we examined the contribution of miR-155 to antitumor immunity using multiple models of syngeneic solid tumor growth in mice. Host miR-155 deficiency inhibited growth of transplanted LLC1 lung tumor cells but not B16 melanoma cells. Interestingly, there was a specific contribution of both T-cell-intrinsic and dendritic cell-intrinsic function of miR-155 positively regulating antitumor T cell immunity. However, host miR-155 deficiency did not lead to overt immune suppression, and instead stimulated robust T-cell- mediated antitumor immunity. On the other hand, the accumulation and function of myeloid-derived suppression cells (MDSC) were impaired in tumor- bearing mice lacking miR-155 and miR155-deficient MDSCs failed to induce T cell antigen unresponsiveness. Moreover, in vivo tumor-inhibitory effect of MDSC depletion by 5-FU was completely abrogated in miR155-deficient mice. Thus, the net antitumor effects of host miR-155 deficiency indicate that the impairment of immunosuppressive MDSCs possibly outweighs the potential suppressive effects of miR-155 deficiency in T cell and dendritic cell compartments. Our study identifies the differential immune regulation for miR-155 during tumor growth, and provides valuable insights for therapeutically intervention in cancer or autoimmune diseases using highly specific methods of modulating miRNAs.

A myosin-Va tail fragment sequesters dynein light chains leading to apoptosis in melanoma cells

Previous studies proposed that myosin-Va regulates apoptosis by sequestering pro-apoptotic Bmf to the actin cytoskeleton through dynein light chain-2 (DLC2). Adhesion loss or other cytoskeletal perturbations would unleash Bmf, allowing it to bind and inhibit pro-survival Bcl2 proteins. Here, we demonstrated that overexpression of a myosin-Va medial tail fragment (MVaf) harboring the binding site for DLC2 dramatically decreased melanoma cell viability. Morphological and molecular changes, including surface blebbing, mitochondrial outer membrane permeabilization, cytochrome-c and Smac release, as well as caspase-9/-3 activation and DNA fragmentation indicated that melanoma cells died of apoptosis. Immobilized MVaf interacted directly with DLCs, but complexed MVaf/DLCs did not interact with Bmf. Overexpression of DLC2 attenuated MVaf-induced apoptosis. Thus, we suggest that, MVaf induces apoptosis by sequestering DLC2 and DLC1, thereby unleashing the pair of sensitizer and activator BH3-only proteins Bmf and Bim. Murine embryonic fibroblasts (MEFs) lacking Bim and Bmf or Bax and Bak were less sensitive to apoptosis caused by MVaf expression than wild-type MEFs, strengthening the putative role of the intrinsic apoptotic pathway in this response. Finally, MVaf expression attenuated B16-F10 solid tumor growth in mice, suggesting that this peptide may be useful as an apoptosis-inducing tool for basic and translational studies.

Control of Solid Tumor Growth in Mice Using EGF Receptor-Targeted RNA Replicase-Based Plasmid DNA

Previously, it was shown that treatment of tumor-bearing mice with an RNA replicase-based plasmid that produces dsRNA when transfected into tumor cells significantly inhibited the tumor growth. In the present study, the feasibility of further improving the anti-tumor activity of the RNA replicase-based plasmid by targeting it into tumors cells was evaluated. An EGF-conjugated, polyethylene glycosylated cationic liposome was developed to deliver the RNA replicase-based plasmid, pSIN-β, into EGF receptor-overexpressing human breast cancer cells (MDA-MB-468) in vitro and in vivo. Delivery of pSIN-β using the EGF receptor-targeted liposome more effectively controlled the growth of MDA-MB-468 tumors (and human epidermoid carcinoma A431 tumors) in mice than using untargeted liposome. The pSIN-β carried by the EGF receptor-targeted liposome caused the complete regression of MDA-MB-468 tumors in mice, probably due to the enhancement of its proapoptotic, antiproliferative and antiangiogenic activities. Tumor-targeted RNA replicase-based plasmids hold a strong potential in tumor therapy.

The calcineurin B subunit induces TNF-related apoptosis-inducing ligand (TRAIL) expression via CD11b–NF-κB pathway in RAW264.7 macrophages

We showed previously that calcineurin B subunit (CnB) could inhibit S180 solid tumor growth in mice and prolong the survival of mice bearing H22 ascites tumors, but the underlying antitumor mechanism remained unclear. Here, we report that the calcineurin B subunit binds to CD11b on RAW264.7 macrophages and induces TRAIL expression and NF-κB activation in a dose and time dependent manner, and that CnB-induced TRAIL expression and NF-κB activation are both dependent on this CD11b. Furthermore, CnB-induced TRAIL expression is mediated by NF-κB. These findings reveal a novel signaling pathway (CnB–CD11b–NF-κB–TRAIL) regulating TRAIL expression and may help to understand the roles of the calcineurin B subunit in the regulation of innate immunity.

Suppression of Human Solid Tumor Growth in Mice by Intratumor and Systemic Inoculation of Histidine-Rich and pH-Dependent Host Defense–like Lytic Peptides

Previously, we reported that intratumor or systemic inoculation of a cationic 15-mer, innate immunity-like lytic peptide composed of d- and l-amino acids ([D]-K(6)L(9)) caused growth arrest of 22RV1 prostate carcinoma xenografts in a mouse model. However, despite its therapeutic potential, this peptide has significant systemic toxicity at concentrations slightly higher than the therapeutic one. Here, we used the acidic environment created by solid tumors as a trigger to activate anticancer lytic peptides by making them cationic only at low pH levels. We achieved this selectivity by substituting lysines (pKa, approximately 10.5) for histidines (pKa, approximately 6.1) in the parental peptide [D]-K(6)L(9). Histidine is protonated below pH 7. For that purpose, we replaced either three or all six lysines in the parental peptide with histidines to obtain the peptides [D]-K(3)H(3)L(9) and [D]-H(6)L(9). Interestingly, in vitro experiments showed pH-dependent activity only with [D]-H(6)L(9) mainly toward cancer cell lines. However, both peptides showed reduced systemic toxicity compared with the parental peptide. Intratumor and systemic inoculation of these peptides resulted in a significant decrease in the 22RV1 prostate cancer tumor volume and systemic secretion of prostate-specific antigen in a xenograft mice model. Moreover, histologic modifications revealed a significant reduction in new blood vessels selectively in tumor tissues after treatment with the peptides compared with the untreated tumors. The lytic mode of action of these new peptides, which makes it difficult for the cancer cells to develop resistance, and their selective and pH-dependent activity make them potential candidates for treatment of solid cancer tumors.

A Chemical Genetic Screen Identified Small Molecules Modulating Zebrafish Vascular Development and Angiogenesis.

The zebrafish has emerged as an important model for studying vascular development and angiogenesis. Advantages over other models include rapid embryonic development and optical clarity of the embryos. Abnormal angiogenesis is linked to over 70 health conditions and inhibition of angiogenesis is an excellent target for cancer therapy (including hematopoietic malignancies) as tumor growth requires new blood vessels. In this study, we performed a chemical genetic screen in developing zebrafish embryos to identify compounds that modulate zebrafish vascular development and angiogenesis. A zebrafish transgenic line with the Flk1 promoter controlling the GFP (green fluorescent protein) reporter was used. The entire vascular network of the Flk1:GFP fish was marked by GFP and could be visualized under fluorescence microscopy. Screens were performed in 96 well plates with three embryos/well. The outer membranes of healthy zebrafish embryos expressing green fluorescent vasculature were removed by enzymatic digestion with protease at the 16–18 somite stages. Small molecules/compounds were then added to the water at concentrations of 0.1 μM, 1 μM, and 5 μM for 72 hrs. After 24, 48 and 72 hrs of exposure to the compound, the embryos were visually inspected for viability, gross morphological defects, heartbeating rate and circulation. The known angiogenic inhibitor, PD173074 was used as control. An initial screening of 780 compounds in a small molecule compound library identified four small molecules with potent activities in inhibiting zebrafish vascular development. Notably, 1–5 μM of Hit #1 inhibited the growth of cranial vessels and disrupted vascular patterning, which resulted in uneven spacing between intersegmental vessels. While Hit #2 and #5 also inhibited the growth of cranial vessels, the vascular patterning remained unaffected, however, the drug-treated embryos had weak or missing intersegmental vessels. Fish embryos treated by Hit #3 had an enlarged heart and thinner vessels. Interestingly, this screen also identified one compound (Hit #4) with pro-angiogenic activity. Embryos treated with Hit #4 had increased numbers of intersegmental vessels. Hit #2 is a known inhibitor of c-Jun N-terminal kinase and angiogenesis (SP600125), was recently reported to inhibit the proliferation and migration of human endothelial cells in vitro as well as inhibit solid tumor growth in mice. This observation lends validity to the zebrafish screen and the potential utility of the other hits. Experiments are currently in progress to investigate the detailed time course of the angiogenic inhibition and the potential molecular mechanism. Studies of these angiogenic inhibitors may lead to the development of potent anti-cancer drugs while the pro-angiogenic compound may prove useful in facilitating tissue/organ regeneration. We conclude that the zebrafish model is likely to yield valuable information regarding vasculogenesis and its manipulation.

Systemic administration of a peptide that impairs the protein kinase (CK2) phosphorylation reduces solid tumor growth in mice

The antitumor efficacy of the CK2 inhibitors so far described has not been extensively evaluated in cancer animal models. We have previously demonstrated that a proapoptotic cyclic peptide termed P15 delivered into the cells by the Tat Cell Penetrating Peptide was able to abrogate the CK2-mediated phosphorylation and induce tumor regression when injected directly into solid tumors in mice. Here we explored the antitumor effect by systemic administration of P15-Tat in a consecutive 5-day schedule through either intraperitoneal or intravenous route. Importantly, significant delay of tumor growth was observed at 2 mg/kg (p < 0.05), 10 mg/kg (p < 0.01) or 40 mg/kg (p < 0.001) after P15-Tat administration both in syngeneic murine tumors and human tumors xenografted in nude mice. In line with this, the systemic administration of P15-Tat induced apoptosis in the tumor as evidenced by in situ DNA fragmentation. Furthermore, we evidenced that 99mTc-labeled P15-Tat peptide was certainly accumulated on the tumors after administration by both routes. This report becomes the first describing the antitumor effect induced by systemic administration of a peptide that targets the acidic phosphorylation domain for CK2 substrates. Also, our data reinforces the perspectives of P15-Tat for the cancer targeted therapy.

Soluble and cell-associated forms of some yet to be identified factor in transfused blood which promotes solid tumor growth in mice.

The mechanism underlying the immunomodulation caused by blood transfusion has yet to be elucidated. The aim of the present study was to determine whether the transfusion of a soluble or insoluble factor present in stored blood can induce immunomodulation, which would thereby promote solid tumor growth. C57Bl/6J mice were subcutaneously inoculated with B16-CG melanoma cells, which secrete beta-human chorionic gonadotropin (beta-hCG). Following inoculation, each of three different products of allogeneic and syngeneic blood were transfused on days 0 and 1: fresh whole blood, stored whole blood, and supernatants from the stored blood. Tumor growth was then monitored by measuring urinary beta-hCG. All mice were killed on day 15, and the tumor weight and volume were measured. Transfusion of all allogeneic blood products enhanced tumor growth, as did the stored syngeneic whole blood. Neither fresh syngeneic blood nor the supernatant from stored syngeneic blood promoted tumor growth. Although the tumors were not visually detectable until day 10 after inoculation, by day 7 the levels of urinary beta-hCG were significantly higher in the mice that received allogeneic blood supernatant than in the mice that received saline. A soluble alloantigen enhances solid tumor growth, as does an insoluble factor present in stored syngeneic whole blood. The immunomodulation associated with this factor begins to enhance tumor growth within 7 days after transfusion.

Inhibitory Effects of Salmosin, a Disintegrin, on Posterior Capsular Opacification in vitro and in vivo

The proliferation, migration and transdifferentiation of the remaining lens epithelial cells (LECs) after cataract surgery are a major cause of posterior capsular opacification (PCO). It has previously been reported that salmosin, a novel disintegrin, significantly inhibits solid tumor growth in mice by perturbation of tumor-specific angiogenesis via blocking αvβ3 integrin expressed on vascular endothelial cells. In this study, the inhibitory function of salmosin in PCO was investigated and was found that salmosin inhibits the attachment of bovine LECs and rabbit lens cells (N/N1003A) to extracellular matrix-coated plates. The anti-adhesive activity of salmosin was approximately 1000 times higher than that of synthetic Arg–Gly–Asp peptide. In addition, the cell proliferation and migration of bovine LECs and N/N1003A were strongly inhibited by salmosin, whereas the proliferation of corneal endothelial cells was less affected. LEC migration and proliferation were also decreased by salmosin treatment in rabbit eyes without any toxic effect in the cornea, iris and retina. In this study, salmosin was shown to specifically inhibit LEC migration and proliferation in an animal model. Therefore, the authors suggest that further investigation may show salmosin to be a good candidate for inhibiting PCO development.

Suppressive Mechanism of Salmosin, a Novel Disintegrin in B16 Melanoma Cell Metastasis

We have previously reported that salmosin, a novel disintegrin, was isolated from Korean snake (Agkistrodon halys brevicaudus) venom and significantly inhibited solid tumor growth in mice by perturbation of tumor-specific angiogenesis via blocking αvβ3 integrin expressed on vascular endothelial cells. In this study, we investigated the functional specificity of salmosin in tumor cell metastasis. Recombinant salmosin expressed in E. coli that has the RGD sequence markedly inhibited both B16F10 melanoma cell adhesion to the extracellular matrix proteins as well as B16F10 melanoma cell invasion through Matrigel-coated filter. The inhibition by salmosin can be caused by blocking integrins expressed on the surface of B16F10 melanoma cells. Salmosin significantly inhibited the proliferation of B16F10 melanoma cells on the plate coated with collagen I in a dose-dependent manner. In vivo B16F10 melanoma experimental metastasis, salmosin showed remarkable significant inhibitory effect on lung tumor colonization in a concentration-dependent manner. These results clearly demonstrate that antimetastatic activity of salmosin resulted from blocking the integrin-mediated adherence and αvβ3 integrin-mediated proliferation of the melanoma cells.

Phase I Study of Immunotherapy of Malignant Melanoma by Direct Gene Transfer. Arizona Cancer Center, Tucson, Arizona

Human Gene TherapyVol. 5, No. 11 News and CommentsPhase I Study of Immunotherapy of Malignant Melanoma by Direct Gene Transfer. Arizona Cancer Center, Tucson, ArizonaPRINCIPAL INVESTIGATOR Evan M. Hersh, CO-PRINCIPAL INVESTIGATORS Emmanuel Akporiaye, David Harris, Alison T. Stopeck, Evan C. Unger, and James A. WarnekePRINCIPAL INVESTIGATOR Evan M. HershSearch for more papers by this author, CO-PRINCIPAL INVESTIGATORS Emmanuel AkporiayeSearch for more papers by this author, David HarrisSearch for more papers by this author, Alison T. StopeckSearch for more papers by this author, Evan C. UngerSearch for more papers by this author, and James A. WarnekeSearch for more papers by this authorPublished Online:19 Mar 2008https://doi.org/10.1089/hum.1994.5.11-1371AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail FiguresReferencesRelatedDetailsCited byDevelopments in targeted therapy in melanomaEuropean Journal of Surgical Oncology (EJSO), Vol. 43, No. 3Control of solid tumor growth in mice using EGF receptor-targeted RNA replicase-based plasmid DNANanomedicine, Vol. 7, No. 4Current progress of DNA vaccine studies in humans9 January 2014 | Expert Review of Vaccines, Vol. 7, No. 2GENE THERAPY FOR MELANOMA IN HUMANSHematology/Oncology Clinics of North America, Vol. 12, No. 3Quantification of Plasmid DNA Expression in VivoClinical Approaches to Cancer Gene TherapyCurrent Therapy Strategies for Malignant Melanoma with Special Regard to Immunotherapy with CytokinesThe Immunological Basis for the Development of Tumor Cell- and Peptide-Based Vaccines for Treatment of Patients with Renal Cell CarcinomaGene Therapy in the United States: A Five-Year Status Report Gail Ross, Robert Erickson, Debra Knorr, Arno G. Motulsky, Robertson Parkman, Jude Samulski, Stephen E. Straus, and Brian R. Smith20 March 2008 | Human Gene Therapy, Vol. 7, No. 14Converting an alcohol to an amine in a cationic lipid dramatically alters the co-lipid requirement, cellular transfection activity and the ultrastructure of DNA-cytofectin complexesBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1280, No. 1Liposome-Mediated Delivery of Genes and Oligonucleotides for the Treatment of Brain TumorsChapter 23. Gene Therapy: Progress, New Directions, and Issues Volume 5Issue 11Nov 1994 InformationCopyright 1994, Mary Ann Liebert, Inc.To cite this article:PRINCIPAL INVESTIGATOR Evan M. Hersh, CO-PRINCIPAL INVESTIGATORS Emmanuel Akporiaye, David Harris, Alison T. Stopeck, Evan C. Unger, and James A. Warneke.Phase I Study of Immunotherapy of Malignant Melanoma by Direct Gene Transfer. Arizona Cancer Center, Tucson, Arizona.Human Gene Therapy.Nov 1994.1371-1384.http://doi.org/10.1089/hum.1994.5.11-1371Published in Volume: 5 Issue 11: March 19, 2008PDF download