Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 7 de 7
Filter
Add more filters










Database
Language
Publication year range
1.
J Cell Biochem ; 108(4): 832-8, 2009 Nov 01.
Article in English | MEDLINE | ID: mdl-19693774

ABSTRACT

The AKT pathway is an important therapeutic target for cancer drug discovery as it functions as a main point for transducing extracellular and intracellular oncogenic signals. Moreover, alternations of the AKT pathway have been found in a wide range of cancers. In the present study, we found that an Akt1 antisense oligonucleotide (Akt1 AO) significantly downregulated the expression of AKT1 at both the mRNA and protein levels and inhibited cellular growth at nanomolar concentrations in various types of human cancer cells. Combined treatment of Akt1 AO with several cytotoxic drugs resulted in an additive growth inhibition of Caki-1 cells. The in vivo effectiveness of Akt1 AO was determined using two different xenograft nude mouse models. Akt1 AO (30 mg/kg, i.v. every 48 h) significantly inhibited the tumor growth of nude mouse subcutaneously implanted with U251 human glioblastoma cells after 27 days treatment. Akt1 AO (30 mg/kg, i.p continuously via osmotic pump) also significantly inhibited the tumor formation in nude mice implanted with luciferase-expressing MIA human pancreatic cancer cells (MIA-Luc) after 14 days of treatment. The luciferase signals from MIA-Luc cells were reduced or completely abolished after 2 weeks of treatment and the implanted tumors were barely detectable. Our findings suggest that Akt1 AO alone or in combination with other clinically approved anticancer agents should be further explored and progressed into clinical studies as a potential novel therapeutic agent.


Subject(s)
Antineoplastic Agents/pharmacology , Oligonucleotides, Antisense/pharmacology , Proto-Oncogene Proteins c-akt/genetics , Animals , Cell Line, Tumor , Female , HeLa Cells , Humans , Inhibitory Concentration 50 , Mice , Mice, Nude , Neoplasm Transplantation , Neoplasms/therapy , Proto-Oncogene Proteins c-akt/metabolism , RNA, Messenger/metabolism , Xenograft Model Antitumor Assays
2.
J Cell Biochem ; 104(3): 985-94, 2008 Jun 01.
Article in English | MEDLINE | ID: mdl-18275063

ABSTRACT

HIF-1alpha plays a major role in activating gene transcription and is important for maintaining homeostasis under hypoxic conditions. Since tumors are often in a hypoxic state, HIF-1alpha is a potential target for the development of novel cancer therapeutics. This study was performed to determine the antitumoral efficacy of an antisense HIF-1alpha inhibitor, RX-0047 on different human cancer cell lines (MDA-MB 231, HME50-T, PC-3, Panc-1 and A549) in vitro. A549 lung cancer and PC-3 prostate cancer cells containing a luciferase gene reporter were used for in vivo xenograft animal models. Progressive tumor development was quantified using live animal BLI (bioluminescence imaging) in addition to ex vivo imaging and histology. All cell lines tested were sensitive to inhibition of cell growth with 10 nM and higher ranges of RX-0047, additionally RX-0047 sensitizes cells to ionizing radiation treatments. Finally, RX-0047 (30 mg/kg) inhibited the formation of human lung metastasis in xenograft mouse models and reduced tumor size in flank models.


Subject(s)
Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors , Oligonucleotides/pharmacology , Transcription, Genetic , Animals , Antineoplastic Agents/pharmacology , Cell Line , Cell Line, Tumor , Genes, Reporter , Humans , In Vitro Techniques , Inhibitory Concentration 50 , Lung Neoplasms/drug therapy , Mice , Mice, Nude , Models, Chemical , Neoplasm Metastasis , Neoplasm Transplantation
3.
Breast Cancer Res Treat ; 96(1): 73-81, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16319992

ABSTRACT

Telomerase activity is undetectable in most normal tissues but the vast majorities of cancers express active telomerase. Therefore, telomerase serves as an attractive target for the treatment of cancers. GRN163L is a lipid-modified oligonucleotide N3'-->P5' thio-phosphoramidate complementary to the RNA template region of human telomerase. The anti-telomerase activity of GRN163L was evaluated using MDA-MB-231 and MDA-MB-435 human breast adenocarcinoma cell lines. Twice weekly administration of GRN163L resulted in the inhibition of telomerase activity and progressive telomere shortening. Cells treated with GRN163L did not demonstrate decreased cell proliferation for up to 2 weeks. However, after additional treatment, cell proliferation gradually decreased in GRN163L-treated cells compared to untreated or mismatch control oligoncleotide treated cells. Furthermore, anti-tumorigenic effects were seen in cells treated with GRN163L, as cells lose their ability to form colonies in soft agar and were unable to form colonies in the clonal efficiency assay upon incubation with GRN163L. Moreover, breast cancer cells that were treated with GRN163L for only 1 week prior to plating in invasion chambers, and when bulk telomere are still long, exhibit significantly diminished invasive potential. These results reveal critical information regarding the effectiveness of GRN163L as a potential therapeutic agent for the treatment of human breast cancer.


Subject(s)
Adenocarcinoma/drug therapy , Breast Neoplasms/drug therapy , Oligopeptides/pharmacology , Telomerase/antagonists & inhibitors , Adenocarcinoma/genetics , Adenocarcinoma/pathology , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Cell Line, Tumor/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Collagen/metabolism , Colony-Forming Units Assay , Drug Combinations , Female , Humans , Laminin/metabolism , Neoplasm Invasiveness/prevention & control , Oligonucleotides , Proteoglycans/metabolism , Telomerase/metabolism , Telomere/genetics , Telomere/metabolism
4.
Cancer Res ; 65(17): 7866-73, 2005 Sep 01.
Article in English | MEDLINE | ID: mdl-16140956

ABSTRACT

Differential regulation of telomerase activity in normal and tumor cells provides a rationale for the design of new classes of telomerase inhibitors. The telomerase enzyme complex presents multiple potential sites for the development of inhibitors. GRN163L, a telomerase enzyme antagonist, is a lipid-modified 13-mer oligonucleotide N3' --> P5'-thio-phosphoramidate, complementary to the template region of telomerase RNA (hTR). We evaluated both the in vitro and in vivo effects of GRN163L using A549-luciferase (A549-Luc) human lung cancer cells expressing a luciferase reporter. GRN163L (1 micromol/L) effectively inhibits telomerase activity of A549-Luc cells, resulting in progressive telomere shortening. GRN163L treatment also reduces colony formation in soft agar assays. Surprisingly, after only 1 week of treatment with GRN163L, A549-Luc cells were unable to form robust colonies in the clonal efficiency assay, whereas the mismatch control compound had no effect. Finally, we show that in vivo treatment with GRN163L is effective in preventing lung metastases in xenograft animal models. These in vitro and in vivo data support the development of GRN163L as a therapeutic for the treatment of cancer.


Subject(s)
Enzyme Inhibitors/administration & dosage , Lung Neoplasms/drug therapy , Oligonucleotides/administration & dosage , Telomerase/antagonists & inhibitors , Animals , Cell Line, Tumor , Enzyme Inhibitors/pharmacokinetics , Humans , Liposomes , Luciferases/metabolism , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Lung Neoplasms/secondary , Mice , Mice, Nude , Neoplastic Stem Cells/drug effects , Oligonucleotides/pharmacokinetics , Sulfur Radioisotopes , Tissue Distribution , Xenograft Model Antitumor Assays
5.
Oncogene ; 24(33): 5262-8, 2005 Aug 04.
Article in English | MEDLINE | ID: mdl-15940257

ABSTRACT

The vast majority of human cancers express telomerase activity, while most human somatic cells do not have detectable telomerase activity. Since telomerase plays a critical role in cell immortality, it is an attractive target for a selective cancer therapy. Oligonucleotides complementary to the RNA template region of human telomerase (hTR) have been shown to be effective inhibitors of telomerase and, subsequently, cancer cell growth in vitro. We show here that a lipid-modified N3'-->P5' thio-phosphoramidate oligonucleotide (GRN163L) inhibits telomerase more potently than its parental nonconjugated thio-phosphoramidate sequence (GRN163). Cells were treated with both the first- (GRN163) and second-generation (GRN163L) oligonucleotides, including a mismatch control, with or without a transfection enhancer reagent. GRN163L inhibited telomerase activity effectively in a dose-dependent manner, even without the use of a transfection reagent. The IC50 values for GRN163 in various cell lines were on average sevenfold higher than for GRN163L. GRN163L inhibition of telomerase activity resulted in a more rapid loss of telomeres and cell growth than GRN163. This report is the first to show that lipid modification enhanced the potency of the novel GRN163 telomerase inhibitor. These results suggest that the lipid-conjugated thio-phosphoramidates could be important for improved pharmacodynamics of telomerase inhibitors in cancer therapy.


Subject(s)
Oligonucleotides/pharmacology , Telomerase/antagonists & inhibitors , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Humans , Lipids/chemistry , Mice , Mice, Nude , Oligonucleotides/chemistry , Solubility , Structure-Activity Relationship , Telomerase/metabolism , Tumor Cells, Cultured/enzymology
6.
Biochem Biophys Res Commun ; 315(4): 1025-32, 2004 Mar 19.
Article in English | MEDLINE | ID: mdl-14985115

ABSTRACT

The urokinase-type plasminogen activator receptor (uPAR) serves as a receptor for urokinase plasminogen activator (uPA) and plays a role in invasion and migration of certain immune cells, including NK cells. Although uPAR is anchored to the plasma membrane via a glycosylphosphatidylinositol lipid moiety, we have previously shown that uPAR crosslinking results in MAP kinase signaling and increased integrin expression on the surface of the human NK cell line, YT. We report, herein, that the binding of uPA to uPAR also activates the MAP kinase signaling cascade. Furthermore, we show the physical association between uPAR and integrins on YT cells using cocapping and fluorescence microscopy. These results suggest that signaling initiated by either uPAR binding to uPA or by uPAR clustering may depend on the physical association of uPAR with integrins, a process that may be a prerequisite for NK cell accumulation within established tumor metastases during adoptive therapy.


Subject(s)
Cell Membrane/metabolism , Integrin alphaV/metabolism , Killer Cells, Natural/metabolism , Receptors, Cell Surface/metabolism , Acetylglucosamine/pharmacology , Flow Cytometry , Fluorescent Antibody Technique , Humans , Immunoblotting , Integrin alphaV/chemistry , Killer Cells, Natural/enzymology , MAP Kinase Signaling System , Microscopy, Fluorescence , Mitogen-Activated Protein Kinases/metabolism , Receptors, Cell Surface/antagonists & inhibitors , Receptors, Cell Surface/chemistry , Receptors, Urokinase Plasminogen Activator , Urokinase-Type Plasminogen Activator/metabolism
7.
J Cell Biochem ; 89(2): 279-88, 2003 May 15.
Article in English | MEDLINE | ID: mdl-12704791

ABSTRACT

Urokinase-type plasminogen activator receptor (uPAR) is attached to cell membranes by a glycosylphosphatidylinositol (GPI) anchor, and as such is devoid of an intracellular domain, but is nevertheless able to initiate signal transduction. Herein, we report a relationship between integrins and uPAR on the surface of the human NK cell line, YT. Our data reveals that crosslinking uPAR, which mimics uPAR clustering at focal adhesion sites, causes increases in expression of the alpha(M), alpha(V), and beta(2) integrins on the surface of YT cells. Activation of the MEK/ERK signaling cascade occurs following uPAR crosslinking, as phosphorylation of both MEK 1/2 and ERK 1/2 results from receptor clustering. The MEK-specific inhibitors PD98059 and U0126 blocked MAP kinase phosphorylation; furthermore, PD98059 inhibited the increase in integrin expression induced by uPAR clustering. This study suggests that uPAR is a signaling receptor and regulator of integrins in NK cells and may impact NK cell function, including the potential for their accumulation within tumor metastases following adoptive transfer.


Subject(s)
Integrins/metabolism , Killer Cells, Natural/metabolism , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinases/metabolism , Receptors, Cell Surface/metabolism , Blotting, Western , Cell Line , Enzyme Activation , Flavonoids/pharmacology , Flow Cytometry , Humans , Mitogen-Activated Protein Kinase 3 , Receptors, Urokinase Plasminogen Activator
SELECTION OF CITATIONS
SEARCH DETAIL
...