lactoferrin and Carcinoma--Squamous-Cell

10 pleomorphic adenomas of the human parotid gland were transplanted on several groups of nude mice. For comparative reasons, 10 other pleomorphic adenomas, a neurinoma and a chordoma and transplants of squamous cell carcinomas and of normal salivary gland tissue were also analysed. In the primary tumours and in the transplants, the presence of keratin, carcinoembryonic antigen, tissue polypeptide antigen, lactoferrin, lysozyme, immunoglobulins, secretory component, amylase, fibronectin and of several lectin-receptors (PNA, WGA, HPA, Ulex europaeus) was sought. The immunohistological observations show that many of the features of a pleomorphic adenoma are constant under the conditions of transplantation. In the transplanted tumour, the same heterogeneity as in the primary tumours can be observed. Autoradiographic studies show little labelling with 3-H thymidine, which is in good accordance with the biological behaviour of the tumour. The distribution of fibronectin shows an interesting association with myoepithelial-like cells. Our results support the hypothesis that the histogenetic origin of the pleomorphic adenoma is a cell pool of the terminal ductal segment. A differentiation towards ductal cells (with production of secretory substances) and towards myoepithelial cells (associated with large amounts of basal membrane like substances) is observed.

Topics: Adenoma, Pleomorphic; Animals; Autoradiography; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Cell Division; Fibronectins; Histocytochemistry; Humans; Immunochemistry; Immunoglobulin A; Immunoglobulin A, Secretory; Immunoglobulin G; Immunoglobulin M; Keratins; Lactoferrin; Lectins; Mice; Mice, Nude; Muramidase; Neoplasm Transplantation; Salivary Gland Neoplasms; Tetradecanoylphorbol Acetate; Transplantation, Heterologous

Topics: AMP-Activated Protein Kinases; Autophagy; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Feedback; Humans; Lactoferrin; Lung; Lung Neoplasms; MicroRNAs

Lactoferrin (LF), a member of the transferrin family, recently has been demonstrated to have anticancer effects on various cancers including oral squamous cell carcinoma (OSCC). However, little is known about the underlying mechanisms of its effects on OSCC. Therefore, we aimed to investigate the mechanism of the suppressive effects of bovine LF (bLF) on the growth of OSCC cells.. In the current study, HSC2, HSC3, HSC4 and normal human oral keratinocytes (RT7) cell lines were tested with bLF 1, 10, and 100 μg/ml. The effects and detail mechanisms of bLF on proliferation and apoptosis of cells were investigated using flow cytometry and western blotting.. We found that bLF (1, 10, and 100 μg/ml) induced activation of p53, a tumor suppressor gene, is associated with the induction of cell cycle arrest in G1/S phase and apoptosis in OSCC. Moreover, bLF downregulated the phosphorylation of Akt and activated suppressor of cytokine signaling 3 (SOCS3), thereby attenuating multiple signaling pathways including mTOR/S6K and JAK/STAT3. Interestingly, we revealed that bLF exerted its effect selectively against HSC3 but not on RT7 via different effects on the phosphorylation status of NF-κB and Akt.. This is the first report showing that bLF selectively suppresses proliferation through mTOR/S6K and JAK/STAT3 pathways and induction of apoptosis in OSCC. This study provides important new findings, which might be useful in the prevention and treatment of OSCC.

Topics: Animals; Carcinoma, Squamous Cell; Cattle; Cell Cycle; Cell Line, Transformed; Cell Proliferation; Humans; Lactoferrin; Mouth Neoplasms

Epithelial-to-mesenchymal transition (EMT) is a biological process of invasion and metastasis in cancers, including in oral squamous cell carcinoma (OSCC). However, an effective anticancer drug that directly targets EMT has not yet been discovered. Therefore, we aimed to investigate the repressive effects of bovine lactoferrin (bLF) on EMT to achieve mesenchymal-to-epithelial transition (MET) in OSCC. OSCC cell lines, HOC313 (EMT-induced) and SCCVII (without EMT induction), were treated with bLF. The effects of bLF on EMT in OSCC were identified histologically by haematoxylin and eosin staining and observed morphologically and immunohistochemically using an anti-E-cadherin antibody. Expression levels of E-cadherin and vimentin were investigated using RT-PCR and western blotting. Immuno-expression of E-cadherin was examined in vivo in tumour tissues of C3H/HeN mice, transplanted with SCCVII cells, with or without bLF administration. We found that bLF changed the spindle-like mesenchymal cells to cuboidal-like epithelial cells and enhanced the affinity of membrane-bound E-cadherin in HOC313 cells. The transformation of EMT-MET in HOC313 cells was confirmed by the upregulation of E-cadherin and suppression of vimentin. Moreover, bLF suppressed TWIST expression through downregulation of ERK1/2 phosphorylation. Additionally, the inhibition tumour cell infiltration and increase in E-cadherin expression were observed in xenografts of the mice orally administered with bLF. Thus, based on the results from in vitro and in vivo studies, we concluded that bLF caused the restoration of epithelial properties through MET. Importantly, this finding is novel and is the first report indicating that bLF inhibited EMT and induced MET in OSCC, suggesting that bLF may provide a novel therapeutic strategy in OSCC.

Topics: Animals; Biomarkers, Tumor; Cadherins; Carcinoma, Squamous Cell; Cattle; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Humans; Lactoferrin; Male; MAP Kinase Signaling System; Mice, Inbred C3H; Mouth Neoplasms; Neoplasm Invasiveness; Nuclear Proteins; Twist-Related Protein 1; Vimentin

Oral squamous cell carcinoma is the fifth most common epithelial cancer in the world, and its current clinical treatment has both low efficiency and poor selectivity. Cationic amphipathic peptides have been proposed as new drugs for the treatment of different types of cancer. The main goal of the present work was to determine the potential of LfcinB(20-25)4, a tetrameric peptide based on the core sequence RRWQWR of bovine lactoferricin LfcinB(20-25), for the treatment of OSCC. In brief, OSCC was induced in the buccal pouch of hamsters by applying 7,12-Dimethylbenz(a)anthracene, and tumors were treated with one of the following peptides: LfcinB(20-25)4, LfcinB(20-25), or vehicle (control). Lesions were macroscopically evaluated every two days and both histological and serum IgG assessments were conducted after 5 weeks. The size of the tumors treated with LfcinB(20-25)4 and LfcinB(20-25) was smaller than that of the control group (46.16±4.41 and 33.92±2.74 mm3 versus 88.77±10.61 mm3, respectively). Also, LfcinB(20-25)4 caused acellularity in the parenchymal tumor compared with LfcinB(20-25) and vehicle treatments. Furthermore, our results demonstrated that both LfcinB(20-25)4 and LfcinB(20-25) induced higher degree of apoptosis relative to the untreated tumors (75-86% vs 8%, respectively). Moreover, although the lowest inflammatory response was achieved when LfcinB(20-25)4 was used, this peptide appeared to induce higher levels of IgG antibodies relative to the vehicle and LfcinB(20-25). In addition the cellular damage and selectivity of the LfcinB(20-25)4 peptide was evaluated in vitro. These assays showed that LfcinB(20-25)4 triggers a selective necrotic effect in the carcinoma cell line. Cumulatively, these data indicate that LfcinB(20-25)4 could be considered as a new therapeutic agent for the treatment of OSCC.

Topics: Animals; Apoptosis; Carcinoma, Squamous Cell; Cattle; Cell Proliferation; Humans; Jurkat Cells; Lactoferrin; Mouth Neoplasms; Peptides

Lactotransferrin (LTF) has been confirmed to act as a tumor suppressor in multiple cancers; however, its roles in oral squamous cell carcinoma (OSCC), one of malignant head and neck carcinomas, has not been explored.. Here, the expression of LTF in OSCC tissues and TCA8113 cells was detected with RT-PCR, qPCR, and IHC. And the correlation between LTF expression and OSCC metastasis was assessed. MS-PCR was performed to reveal the methylation status in promoter regions of LTF both in OSCC tissue samples and cells. The influences of 5-Aza-Cdc treatment to the methylation status and expression levels of LTF were also analyzed. At last, the functions of LTF in OSCC progression were demonstrated by MTT analysis, clone formation assay, and cell cycle analysis in TCA8113 cells with forced ectopic expression of LTF.. LTF showed a low or null expression pattern in OSCC tissues and cells, at least partially, due to the hypermethylated status in promoter regions for 5-Aza-Cdc, a methyltransferase inhibitor, could restore the expression of LTF in TCA8113 cells. And the expression level of LTF exhibited a negative correlation with OSCC metastasis.. Re-expression of LTF inhibited the growth, proliferation, as well as cell cycle progression of TCA8113 cells. In conclusion, hypermethylation contributes much to LTF inactivation in OSCC. And LTF can partially reverse the malignant phenotypes of OSCC cells and may be served as a potential target for diagnosis and therapy of OSCC in future.

Topics: Base Sequence; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; DNA Methylation; Gene Silencing; Genes, Tumor Suppressor; Head and Neck Neoplasms; Humans; Immunohistochemistry; Lactoferrin; Molecular Sequence Data; Mouth Neoplasms; Neoplasm Metastasis; Polymerase Chain Reaction; Promoter Regions, Genetic; Squamous Cell Carcinoma of Head and Neck

Several short linear peptides derived from cyclic bovine lactoferricin were synthesized and tested for their cytotoxic effect against the oral cavity squamous-cell carcinoma (OSCC) cell lines CAL27 and SCC15. As a control, an immortalized and nontumorigenic cell line, Het-1A, was used. Linear peptides based on the RRWQWR core sequence showed a moderate cytotoxic effect and specificity towards tumorigenic cells. A tetrameric peptide, LfcinB(20-25)4, containing the RRWQWR motif, exhibited greater cytotoxic activity (>90%) in both OSCC cell lines compared to the linear lactoferricin peptide or the lactoferrin protein. Additionally, this tetrameric peptide showed the highest specificity towards tumorigenic cells among the tested peptides. Interestingly, this effect was very fast, with cell shrinkage, severe damage to cell membrane permeability, and lysis within one hour of treatment. Our results are consistent with a necrotic effect rather than an apoptotic one and suggest that this tetrameric peptide could be considered as a new candidate for the therapeutic treatment of OSCC.

Topics: Animals; Carcinoma, Squamous Cell; Cattle; Cell Line, Tumor; Cell Membrane Permeability; Cytotoxins; Humans; Lactoferrin; Mouth Neoplasms; Necrosis; Peptides

To investigate the effect of lactoferrin on vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression.. Lactoferrin at concentration of 0.006, 0.013, 0.025, 0.050 g/L and blank control groups were included in this study. The gene and protein expression of VEGF, bFGF were examined by RT-PCR and Western blotting.. The RT-PCR and Western blotting assay showed that VEGF mRNA(0.31 +/- 0.08) and protein (0.68 +/- 0.11) in lactoferrin (0.050 g/L) group were significantly lower than in the control group (P < 0.05), and the bFGFmRNA (0.27 +/- 0.10) and protein (0.68 +/- 0.07) in lactoferrin (0.050 g/L) group were also significantly lower than in the control group (P < 0.05).. Lactoferrin could inhibit the expression of VEGF, bFGFmRNA and protein in Tca8113 cells. This effect might be one of the mechanisms for anticancer function of lactoferrin.

Topics: Antineoplastic Agents; Carcinoma, Squamous Cell; Cell Line, Tumor; Dose-Response Relationship, Drug; Down-Regulation; Fibroblast Growth Factor 2; Humans; Lactoferrin; RNA, Messenger; Tongue Neoplasms; Vascular Endothelial Growth Factor A

Human lactoferrin is a naturally occurring glycoprotein that inhibits cancer growth. Our purpose was to evaluate recombinant human lactoferrin as a chemotherapeutic agent against head and neck squamous cell carcinoma.. Controlled experiments both in vitro and in the murine model evaluating both the effect and mechanism of lactoferrin on cancer growth.. In both human and murine cell lines, lactoferrin induced dose-dependent growth inhibition. Using flow cytometric analysis, lactoferrin was shown to induce G(1)-G(0) growth arrest. This arrest seemed to be modulated by down-regulation of cyclin D1. In the in vitro model, luminex data revealed that lactoferrin inhibited cellular release of proinflammatory and prometastatic cytokines, including interleukin-8, interleukin-6, granulocyte macrophage colony-stimulating factor, and tumor necrosis factor-alpha. Lactoferrin up-regulated the cellular activation of nuclear factor-kappaB within 4 h of cellular exposure. In C3h/HeJ mice implanted with SCCVII tumors, orally delivered lactoferrin inhibited tumor growth by 75% compared with control mice. Immunohistochemical analysis of harvested tumors revealed up to 20-fold increases of lymphocytes within treated animals. When mice were depleted of CD3(+) cells, all lactoferrin-induced tumor inhibition was abrogated.. We conclude that human recombinant lactoferrin can inhibit the growth of head and neck squamous cell carcinoma via direct cellular inhibition as well as systemically via immunomodulation. Our data support the study of human lactoferrin as an immunomodulatory compound with therapeutic potential.

Topics: Administration, Oral; Animals; Antineoplastic Agents; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Cytokines; Flow Cytometry; Head and Neck Neoplasms; Humans; Immunohistochemistry; Lactoferrin; Mice; Recombinant Proteins; T-Lymphocytes

Alpha-defensin or human neutrophil peptide-1 (HNP1) is a neutrophil-derived antimicrobial peptide with cytotoxic effects towards cancer cells. Lactoferrin is also stored in human neutrophils and is a glycoprotein involved in mediating cytotoxicity towards tumour cells. This study investigated the sensitivity of normal oral keratinocyte and oral squamous cell carcinoma (OSCC) cells to HNP1 and lactoferrin in various combinations. A concentration of 100 microg/ml HNP1 induced the most significant cytotoxic effect on both normal and OSCC cells. Lactoferrin (12.5, 25 and 250 microg/ml) also significantly induced cell death in OSCC cells after 72 h. Of note, a combination of 10 microg/ml HNP1 and 50 microg/ml lactoferrin induced a differential effect, not observed with either concentration alone, which stimulated proliferation in normal cells, but induced cell death in OSCC cells throughout the study. These results indicate a potentially important co-operative role for HNP1 and lactoferrin in facilitating a selective cytotoxic effect on tumour cells.

Topics: alpha-Defensins; Carcinoma, Squamous Cell; Cell Death; Cells, Cultured; Dose-Response Relationship, Drug; Drug Interactions; Drug Screening Assays, Antitumor; Electrophoresis, Polyacrylamide Gel; Humans; Keratinocytes; Lactoferrin; Mouth Mucosa; Mouth Neoplasms; Tumor Cells, Cultured

Combination chemoprevention using tea polyphenols as one of the components has received growing consideration in recent years. The present study was designed to evaluate the antiproliferative and apoptosis inducing effects of bovine lactoferrin (bLF) and black tea polyphenol (Polyphenon-B: P-B) combination on 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. Topical application of DMBA for 14 weeks induced buccal pouch tumours that showed aberrant expression of cytokeratins, a marker for epithelial carcinomas. This was associated with increased cell proliferation and evasion of apoptosis as revealed by upregulation of proliferating cell nuclear antigen, NF-kappaB, mutant p53, Bcl-2 and downregulation of Bax, Fas and caspase 3 protein expression. Although dietary administration of bLF and Polyphenon-B alone significantly reduced tumour incidence, combined administration of bLF and Polyphenon-B was more effective in inhibiting HBP carcinogenesis by restoring normal cytokeratin expression, inhibiting cell proliferation and inducing apoptosis. These findings suggest that a "designer item" approach will be useful for human oral cancer prevention strategies.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Apoptosis; Carcinoma, Squamous Cell; Cell Proliferation; Cheek; Chemoprevention; Cricetinae; Lactoferrin; Male; Mesocricetus; Mouth Neoplasms; Phenols; Tea

In this work, we investigated the anticancer activity of orally administered recombinant human lactoferrin (rhLF) alone and in combination with chemotherapy in tumor-bearing mice. rhLF inhibited the growth of squamous cell carcinoma (O12) tumors in T cell-immunocompromised nu/nu mice by 80% when administered at 1,000 mg/kg (2.9 g/m2) by oral gavage twice daily for 8 days (p < 0.001). Similar activity was observed in syngeneic, immunocompetent BALB/c mice, where orally administered rhLF (1,000 mg/kg, 2.9 g/m2 once daily) halted the growth of mammary adenocarcinoma TUBO. Oral rhLF (200 mg/kg, 0.57 g/m2) was also used alone and in combination with cis-platinum (5 mg/kg) to treat head-and-neck squamous cell carcinoma in a syngeneic murine model. Monotherapy with oral rhLF or cis-platinum caused 61% or 66% tumor growth inhibition over placebo, respectively. Mice receiving both therapies showed 79% growth inhibition, a statistically significant improvement over each drug alone. We then demonstrated that administration of oral rhLF (300 mg/kg, 0.86 g/m2) to tumor-bearing or naive mice resulted in (i) significantly increased production of IL-18 in the intestinal tract, (ii) systemic NK cell activation and (iii) circulating CD8+ T-cell expansion. These data suggest that oral rhLF is an immunomodulatory agent active against cancer as a single agent and in combination chemotherapy, exerting its systemic effect through stimulation of IL-18 and other cytokines in the gut enterocytes. rhLF has been administered orally to 211 people without a single serious drug-related adverse event. Thus, rhLF shows promise as a safe and well-tolerated novel immunomodulatory anticancer agent.

Topics: Administration, Oral; Animals; Antineoplastic Agents; Carcinoma, Squamous Cell; Cell Division; Cell Line, Tumor; Cisplatin; Drug Synergism; Female; Flow Cytometry; Humans; Killer Cells, Natural; Lactoferrin; Mice; Mice, Inbred BALB C; Mice, Nude; Recombinant Proteins

Papillary adenocarcinoma is extremely rare in the squamous epithelium-lined esophagus. The histopathologic and immunohistochemical characteristics were examined in a composite tumor showing distinct papillary adenocarcinoma and squamous cell carcinoma of the esophagus resected from a 66-year-old man. The esophageal tumor consisted both grossly and histologically of two distinct components: an ulcerative part showing a squamous cell carcinoma, and a polypoid part corresponding to a papillary adenocarcinoma. In addition, the in situ squamous cell carcinoma was contiguous with the esophageal tumor. Mucin secretion was found only in the papillary adenocarcinoma component. Immunohistochemically, tumor cells of the papillary adenocarcinoma component were positive for carcinoembryonic antigen, secretory component, and lactoferrin. These staining patterns were similar to those of the normal esophageal gland proper. These histologic, mucin-histochemical, and immunohistochemical findings suggest that the papillary adenocarcinoma originated from the submucosal esophageal gland and the squamous cell carcinoma from the squamous epithelium lining the esophagus.

Topics: Adenocarcinoma, Papillary; Aged; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Esophageal Neoplasms; Humans; Immunohistochemistry; Lactoferrin; Male; Mucins; Secretory Component

Immunodiffusion technique was used to study the lactoferrin (LF) distribution in human fetal, normal and tumour tissue extracts. If was found that LF concentration in lung tumour extracts was higher than in normal lung one. Indirect immunoperoxidase method revealed LF in alveolar macrophages of normal and tumour lung tissues, in afflux of these cells around the tumour cells, but never--in the original lung cancer cells. It is suggested that LF is not a lung tumour-associated antigen.

Topics: Carcinoma, Squamous Cell; Female; Fetus; Humans; Immunodiffusion; Immunoenzyme Techniques; Lactoferrin; Lactoglobulins; Lung; Lung Neoplasms; Male; Neoplasm Proteins; Pregnancy; Tissue Distribution; Tissue Extracts

Lactoferrin and lysozyme, parts of the non-specific defense system, were studied in normal and diseased parotid glands, using the immunohistochemical PAP-method. 31 normal and inflamed glands were investigated. The presence of lactoferrin and lysozyme was demonstrated in the acinar cells and some duct cells. The amount of these substances was increased in obstructive parotitis. The 52 carcinomas showed a distinct distribution pattern for lactoferrin (positive cases: adenocarcinomas 5 of 8; cystadenocarcinoma: 3 of 5; adenoid cystic carcinomas 2 of 4; salivary duct carcinomas 2 of 3). Some of the carcinomas in pleomorphic adenomas were positive for lactoferrin. Squamous cell carcinomas and anaplastic carcinomas were constantly negative. All carcinomas were negative for lysozyme. These observations are discussed with respect to their physiological and pathological significance.

Topics: Adenocarcinoma; Adenoma, Pleomorphic; Carcinoma; Carcinoma, Squamous Cell; Cystadenocarcinoma; Humans; Lactoferrin; Lactoglobulins; Muramidase; Parotid Neoplasms; Parotitis