Iron Oxide Nanoparticles Carrying 5-Fluorouracil in Combination with Magnetic Hyperthermia Induce Thrombogenic Collagen Fibers, Cellular Stress, and Immune Responses in Heterotopic Human Colon Cancer in Mice

In this study we looked for the main protein pathway regulators which were responsible for the therapeutic impact on colon cancers when combining magnetic hyperthermia with the chemotherapeutic agent 5-fluorouracil (5FU). To this end, chitosan-coated magnetic nanoparticles (MNP) functionalized with 5FU were intratumorally injected into subcutaneous human colon cancer xenografts (HT-29) in mice and exposed to an alternating magnetic field. A decreased tumor growth was found particularly for the combined thermo-chemotherapy vs. the corresponding monotherapies. By using computational analysis of the tumor proteome, we found upregulated functional pathway categories termed “cellular stress and injury”, “intracellular second messenger and nuclear receptor signaling”, “immune responses”, and “growth proliferation and development”. We predict TGF-beta, and other mediators, as important upstream regulators. In conclusion, our findings show that the combined thermo-chemotherapy induces thrombogenic collagen fibers which are able to impair tumor nutrient supply. Further on, we associate several responses to the recognition of damage associated molecular patterns (DAMPs) by phagocytic cells, which immigrate into the tumor area. The activation of some pathways associated with cell survival implies the necessity to conduct multiple therapy sessions in connection with a corresponding monitoring, which could possibly be performed on the base of the identified protein regulators.

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Immune responses in human colon cancer

Gastroenterology ◽

10.1016/0016-5085(78)90461-4 ◽

1978 ◽

Vol 75 (5) ◽

pp. 800-805 ◽

Cited By ~ 5

Author(s):

William V. Hahn ◽

Martin F. Kagnoff ◽

Loren E. Hatlen ◽

Raleigh K. Austin

Keyword(s):

Colon Cancer ◽

Immune Responses ◽

Human Colon ◽

Human Colon Cancer

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Bone morphogenetic protein signaling and growth suppression in colon cancer

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00482.2005 ◽

2006 ◽

Vol 291 (1) ◽

pp. G135-G145 ◽

Cited By ~ 72

Author(s):

Stayce E. Beck ◽

Barbara H. Jung ◽

Antonio Fiorino ◽

Jessica Gomez ◽

Eunice Del Rosario ◽

...

Keyword(s):

Cell Cycle ◽

Colon Cancer ◽

Transcriptional Activity ◽

Human Colon ◽

Growth Suppression ◽

Bmp Signaling ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Colon Cancers ◽

Sw480 Cells

Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-β superfamily, which utilize BMP receptors and intracellular SMADs to transduce their signals to regulate cell differentiation, proliferation, and apoptosis. Because mutations in BMP receptor type IA ( BMPRIA) and SMAD4 are found in the germline of patients with the colon cancer predisposition syndrome juvenile polyposis, and because the contribution of BMP in colon cancers is largely unknown, we examined colon cancer cells and tissues for evidence of BMP signaling and determined its growth effects. We determined the presence and functionality of BMPR1A by examining BMP-induced phosphorylation and nuclear translocation of SMAD1; transcriptional activity via a BMP-specific luciferase reporter; and growth characteristics by cell cycle analysis, cell growth, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide metabolic assays. These assays were also performed after transfection with a dominant negative (DN) BMPR1A construct. In SMAD4-null SW480 cells, we examined BMP effects on cellular wound assays as well as BMP-induced transcription in the presence of transfected SMAD4. We also determined the expression of BMPR1A, BMP ligands, and phospho-SMAD1 in primary human colon cancer specimens. We found intact BMP signaling and modest growth suppression in HCT116 and two derivative cell lines and, surprisingly, growth suppression in SMAD4-null SW480 cells. BMP-induced SMAD signaling and BMPR1A-mediated growth suppression were reversed with DN BMPR1A transfection. BMP2 slowed wound closure, and transfection of SMAD4 into SW480 cells did not change BMP-specific transcriptional activity over controls due to receptor stimulation by endogenously produced ligand. We found no cell cycle alterations with BMP treatment in the HCT116 and derivative cell lines, but there was an increased G1 fraction in SW480 cells that was not due to increased p21 transcription. In human colon cancer specimens, BMP2 and BMP7 ligands, BMPRIA, and phospho-SMAD1 were expressed. In conclusion, BMP signaling is intact and growth suppressive in human colon cancer cells. In addition to SMADs, BMP may utilize SMAD4-independent pathways for growth suppression in colon cancers.

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Immune Responses in Human Colon Cancer. II. Cytotoxic Antibody Detected in Patients' Sera2

JNCI Journal of the National Cancer Institute ◽

10.1093/jnci/60.4.779 ◽

1978 ◽

Vol 60 (4) ◽

pp. 779-784 ◽

Cited By ~ 10

Author(s):

William V. Hahn ◽

Martin F. Kagnoff ◽

Loren H. Hatlen

Keyword(s):

Colon Cancer ◽

Immune Responses ◽

Human Colon ◽

Human Colon Cancer ◽

Cytotoxic Antibody

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Site-specific DNA methylation contributes to neurotensin/neuromedin N expression in colon cancers

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2000.279.6.g1139 ◽

2000 ◽

Vol 279 (6) ◽

pp. G1139-G1147 ◽

Cited By ~ 17

Author(s):

Zizheng Dong ◽

Xiaofu Wang ◽

B. Mark Evers

Keyword(s):

Dna Methylation ◽

Colon Cancer ◽

Oligonucleotide Probe ◽

Consensus Sequence ◽

Human Colon ◽

N Gene ◽

Human Colon Cancer ◽

Cpg Dinucleotides ◽

Colon Cancers ◽

Neuromedin N

The neurotensin/neuromedin N (NT/N) gene is expressed in fetal colon, repressed in newborn and adult colon, and reexpressed in ∼25% of colon cancers. Our purpose was to determine the effect of gene methylation on NT/N silencing in colon cancers. We found that the NT/N gene was expressed in human colon cancer cell line KM12C but not in KM20 colon cancer cells. Bisulfite genomic sequencing demonstrated that all CpG dinucleotides in the region from −373 to +100 of the NT/N promoter, including a CpG site in a distal consensus AP-1 site, were methylated in KM20 but unmethylated in KM12C cells. Treatment of KM20 cells with demethylating agent 5-azacytidine induced NT/N expression, suggesting a role for DNA methylation in silencing of NT/N in colon cancers. To better elucidate the mechanisms responsible for NT/N repression by DNA methylation, we performed gel shift assays using an oligonucleotide probe corresponding to the distal AP-1 consensus sequence of the NT/N promoter. Methylation of the oligonucleotide probe inhibited protein binding to the distal AP-1 site of the NT/N promoter, suggesting a potential mechanism of NT/N gene repression in colon cancers. We show that DNA methylation plays a role in NT/N gene silencing in the human colon cancer KM20 and that NT/N expression in KM12C cells is associated with demethylation of the CpG sites. DNA methylation likely contributes to NT/N gene expression noted in human colon cancers.

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Expression of GRP and Its Receptor in Well-differentiated Colon Cancer Cells Correlates with the Presence of Focal Adhesion Kinase Phosphorylated at Tyrosines 397 and 407

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215540305100807 ◽

2003 ◽

Vol 51 (8) ◽

pp. 1041-1048 ◽

Cited By ~ 35

Author(s):

Kristina A. Matkowskyj ◽

Kristin Keller ◽

Sarah Glover ◽

Lori Kornberg ◽

Roger Tran-Son-Tay ◽

...

Keyword(s):

Colon Cancer ◽

Cell Differentiation ◽

Tumor Cell ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Human Colon ◽

Human Colon Cancer ◽

Colon Cancers ◽

Well Differentiated ◽

Tumor Cell Differentiation

Gastrin-releasing peptide (GRP) and its receptor (GRP-R) are not normally expressed by epithelial cells lining the colon but are aberrantly expressed in cancer, where they act as morphogens and regulate tumor cell differentiation. Studies of colon cancer formation in mice genetically incapable of synthesizing GRP-R suggested that this receptor's morphogenic properties were mediated via focal adhesion kinase (FAK). We therefore set out to determine the presence of both total and phosphorylated forms of FAK in human colon cancer specimens as a function of tumor cell differentiation and GRP/GRP-R co-expression. Ten colon cancers containing 25 regions of distinct differentiation were randomly selected from our GI Cancer Tumor Bank. All specimens were immunohistochemically probed using antibodies recognizing GRP, GRP-R, total FAK, and FAK specifically phos-phorylated at tyrosine (Y) 397, 407, 576, 577, 861, and 925. Antibody-specific chromogen was determined by quantitative immunohistochemistry (IHC) for each region of defined differentiation. Here we confirm that GRP/GRP-R co-expression is a function of differentiation, with highest levels observed in well-differentiated tumor cells. We also show that the amount of total FAK and of FAK phosphorylated at Y397 and Y407 tightly correlates with differentiation and with the amount of GRP/GRP-R co-expression. These findings are consistent with GRP/GRP-R acting as a morphogen by activating FAK, and suggest that this occurs via phosphorylation of this enzyme at two specific tyrosine residues.

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