elastin and Carcinoma--Squamous-Cell

To present the pathologic analysis of female urethral strictures obtained during reconstructive urethroplasty.. Nine separate female urethral tissue specimens were obtained during dorsal vaginal graft urethroplasty by a single surgeon (S.P.P.). Samples were serially sectioned and fixed in 10% formalin 6 to 12 hours before routine processing in paraffin blocks. Serial 5-µm sections were subjected to H&E, Masson trichrome, and elastin staining. End point analysis included evaluation for epithelial hyperplasia and cell type, mucosal edema, degree of fibroblast/inflammatory cell infiltrate, and elastin fiber density and distribution.. Nine specimens were examined. Six specimens had epithelial linings of stratified squamous epithelium overlying fibrosis (67%), 1 had mixed squamous and urothelial epithelium, and 2 had only urothelial epithelium. Two specimens (29%) showed acute injury with prominent squamous papillary hyperplasia, focal erosion, and patchy mucosal hemorrhage. Areas of urethral stricture were variably thickened, with increased, densely packed collagen fibers and associated mucosal lymphocytic inflammation ranging from mild and patchy to focally dense with lymphoid aggregates. The highest elastin fiber density appeared to be associated with vessels and overlying muscle bundles in the submucosa.. Further elucidation of histopathologic characteristics may illuminate more appropriate therapeutic pathways for female urethral stricture disease management.

Topics: Carcinoma, Squamous Cell; Elastin; Female; Humans; Hyperplasia; Male; Mouth Mucosa; Treatment Outcome; Urethral Stricture; Urothelium

During the pathological destruction of lung tissue, neutrophil elastase (NE) degrades elastin, one of the major constituents of lung parenchyma. However there are no non-invasive methods to quantify NE degradation of elastin. We selected specific elastin fragments generated by NE for antibody generation and developed an ELISA assay (EL-NE) for the quantification of NE-degraded elastin.. Monoclonal antibodies were developed against 10 NE-specific cleavage sites on elastin. One EL-NE assay was tested for analyte stability, linearity and intra- and inter-assay variation. The NE specificity was demonstrated using elastin cleaved in vitro with matrix metalloproteinases (MMPs), cathepsin G (CatG), NE and intact elastin. Clinical relevance was assessed by measuring levels of NE-generated elastin fragments in serum of patients diagnosed with idiopathic pulmonary fibrosis (IPF, n = 10) or lung cancer (n = 40).. Analyte recovery of EL-NE for human serum was between 85% and 104%, the analyte was stable for four freeze/thaw cycles and after 24 h storage at 4°C. EL-NE was specific for NE-degraded elastin. Levels of NE-generated elastin fragments for elastin incubated in the presence of NE were 900% to 4700% higher than those seen with CatG or MMP incubation or in intact elastin. Serum levels of NE-generated elastin fragments were significantly increased in patients with IPF (137%, p = 0.002) and in patients with lung cancer (510%, p < 0.001) compared with age- and sex-matched controls.. The EL-NE assay was specific for NE-degraded elastin. The EL-NE assay was able to specifically quantify NE-degraded elastin in serum. Serum levels of NE-degraded elastin might be used to detect excessive lung tissue degradation in lung cancer and IPF.

Topics: Adenocarcinoma; Aged; Carcinoma, Squamous Cell; Case-Control Studies; Elastin; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; Female; Humans; Idiopathic Pulmonary Fibrosis; Leukocyte Elastase; Lung; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Small Cell Lung Carcinoma

Silk-elastinlike protein polymers (SELPs) have been effectively used as controlled release matrices for the delivery of viruses for cancer gene therapy in preclinical models. However, the degradability of these polymers needs to be tuned for improved localized intratumoral gene delivery. Using recombinant techniques, systematic modifications in distinct regions of the polymer backbone, namely, within the elastin blocks, silk blocks, and adjacent to silk and elastin blocks, have been made to impart sensitivity to specific matrix metalloproteinases (MMPs) known to be overexpressed in the tumor environment. In this report we investigated the structure-function relationship of MMP-responsive SELPs for viral mediated gene therapy of head and neck cancer. These polymers showed significant degradation in vitro in the presence of MMPs. Their degradation rate was a function of the location of the MMP-responsive sequence in the polymer backbone when in hydrogel form. Treatment efficacy of the adenoviral vectors released from the MMP responsive SELP analogs in a xenograft mouse model of head and neck squamous cell carcinoma (HNSCC) was shown to be polymer structure dependent. These results demonstrate the tunable nature of MMP-responsive SELPs for localized matrix-mediated gene delivery.

Topics: Adenoviridae; Animals; Carcinoma, Squamous Cell; Cell Line, Tumor; Delayed-Action Preparations; Elastin; Female; Gene Transfer Techniques; Genetic Therapy; Genetic Vectors; Head and Neck Neoplasms; Humans; Matrix Metalloproteinases; Mice, Nude; Silk; Squamous Cell Carcinoma of Head and Neck

Epithelial changes observed in actinic cheilitis (AC) and lower lip squamous cell carcinoma (LLSCC) have been studied using different markers in order to observe diagnostic and prognostic factors for both lesions. The aim of the present study was to analyze Ki-67, TGF-β1, and elastin content in AC and LLSCC to determine the possible role of these proteins in lip carcinogenesis. Medical records of 29 cases of AC and 53 cases of LLSCC were analyzed. Lesions were classified according histological pattern and submitted to immunostaining for Ki-67, TGF-β1, and elastin. Different percentages of Ki-67-positive cells were found in AC depending on the degree of epithelial dysplasia (p < 0.01). An association was also found between the percentage of Ki-67-positive cells and tumor grade in LLSCC (p < 0.01). An inverse correlation was found between Ki-67 and TGF-β1 in AC and LLSCC (p < 0.01). Elastosis was thinner and more discontinuous in LLSCC in comparison to AC, and this difference in the elastin immunolabeling pattern was statistically significant between groups (p < 0.01). The present findings indicate that changes in Ki-67 and TGF-β1 content contribute to lip carcinogenesis. Furthermore, elastin content reflects changes in the extracellular matrix in both AC and LLSCC.

Topics: Adult; Aged; Carcinogenesis; Carcinoma, Squamous Cell; Elastin; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Lip Neoplasms; Logistic Models; Male; Middle Aged; Transforming Growth Factor beta1

Brachytherapy is a common clinical technique involving implantation of sealed radioactive "seeds" within a tumor to selectively irradiate the tumor mass while minimizing systemic toxicity. To mitigate the disadvantages associated with complex surgical implantation and subsequent device removal procedures, we have developed an alternative approach using a genetically encoded peptide polymer solution composed of a thermally responsive elastin-like polypeptide (ELP) radiolabeled with (131)I that self-assembles into radionuclide seeds upon intratumoral injection. The formation of these nontoxic and biodegradable polymer seeds led to prolonged intratumoral retention (~85% ID/tumor 7 days postinjection) of the radionuclide, elicited a tumor growth delay in 100% of the tumors in two human xenografts (FaDu and PC-3), and cured more than 67% of tumor-bearing animals after a single administration of labeled ELP. These results suggest that in situ self-assembly of biodegradable and injectable radionuclide-containing polypeptide seeds could be a promising therapeutic alternative to conventional brachytherapy.

Topics: Animals; Brachytherapy; Carcinoma, Squamous Cell; Cell Line, Tumor; Drug Delivery Systems; Elastin; Female; Humans; Hypopharyngeal Neoplasms; Iodine Radioisotopes; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Peptides; Prostatic Neoplasms; Tandem Repeat Sequences; Xenograft Model Antitumor Assays

Mast cells (MCs) display a diversity of roles that may contribute to the stromal microenvironment alterations during tumor progression. The aim of this study was to investigate MC populations expressing tryptase and c-kit in lip squamous cell carcinoma (lip SCC) (n=37), actinic cheilitis (AC) (n=15) and normal lip mucosa (control) (n=6), as well as their relationship with microscopic parameters (collagen degeneration, elastin changes, angiogenesis and proliferative index). Tryptase, c-kit, CD31 and Ki-67 expressions were analyzed by means of immunohistochemistry and collagen and elastic fibers were visualized with Picrosirus and Verhoeff's stain, respectively. The numbers of tryptase+ MC were significantly higher in lip SCC when compared with control (P=0.01), while a similar density of these cells was observed in AC and lip SCC (P=0.09). The density of c-kit+ MC was similar in all groups examined (P=0.65). MC migration (c-kit+/Tryptase+ relationship) was 69% in lip SCC, 60% in AC and 100% in control. The number of CD31+ blood vessels was significantly higher in the lip SCC when compared with control and AC (P<0.01). The increase of MCs and angiogenesis in lip SCC may reflect an important modification in the tumor microenvironment during squamous photo-carcinogenesis.

Topics: Adult; Age Factors; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Movement; Cheilitis; Collagen; Elastin; Extracellular Matrix; Female; Humans; Ki-67 Antigen; Lip Neoplasms; Male; Mast Cells; Middle Aged; Mouth Mucosa; Neovascularization, Pathologic; Platelet Endothelial Cell Adhesion Molecule-1; Proto-Oncogene Proteins c-kit; Tryptases; Ultraviolet Rays

To investigate the expression and significance of elastin and fibulin-5 in anterior vaginal tissue of women with pelvic organ prolapse (POP).. Between November 2006 and June 2008, 68 patients with POP underwent surgical treatment in Shengjing Hospital of China Medical University were enrolled in this study, who were classified into 10 patients with grade I, 21 patients with grade II, 25 patients with grade III and 12 patients with grade IV in accordance with pelvic organ prolapse quantitation (POP-Q). Meanwhile, 18 cases with early cervical cancer at stage of I b were treated by total hysterectomy and bilateral salpingo-oophorectomy, their anterior vaginal tissues were selected as controls. Immunohistochemical staining was performed to detect the expression of elastin and fibulin-5.. (1) Elastin and fibulin-5 were mainly expressed at extracellular matrix(ECM). (2) The positive rate of fibulin-5 expression in anterior vaginal wall were 5% (2/37) in grade III/IV and 26% (8/31) in grade I/IV POP patients, which reached statistical difference (P = 0.035). However, no statistical different expression was found between postmenopausal (13%, 8/60) and non-menopausal patients (2/8), vaginal delivery < or =2 (19%, 5/27) and >2 patients (12%, 5/41, P > 0.05). (3) The positive rate of elastin expression in anterior vaginal wall in POP group was 31% (21/68), which was significantly lower than 72% (13/18) of control group (P = 0.002). Among POP group, 19% (7/37) of elastin expression in grade III/IV POP was significantly lower than 45% (14/31) in grade I/II of POP patients. However, no statistical difference was found between postmenopausal (30%, 18/60) and non-menopausal patients (3/8), vaginal delivery < or =2 (26%, 7/27) and >2 patients(34%, 14/41, P > 0.05). (4) In POP group, both positive expression of fibulin-5 and elastin of anterior vaginal wall was in 6 cases, both negative expression of fibulin-5 and elastin was in 43 cases. It was illustrated that elastin and fibulin-5 had an positive relationship (P = 0.031).. The decreased expression of elastin and fibulin-5 was correlated with degree of POP, which indicated that elastin and fibulin-5 may play a role in the pathogenesis of POP.

Topics: Aged; Carcinoma, Squamous Cell; Connective Tissue; Elastin; Extracellular Matrix Proteins; Female; Humans; Immunohistochemistry; Menopause; Middle Aged; Pelvic Organ Prolapse; Severity of Illness Index; Uterine Cervical Neoplasms; Vagina

The delivery of anticancer therapeutics to solid tumors remains a critical problem in the treatment of cancer. This study reports a new methodology to target a temperature-responsive macromolecular drug carrier, an elastin-like polypeptide (ELP) to solid tumors. Using a dorsal skin fold window chamber model and intravital laser scanning confocal microscopy, we show that the ELP forms micron-sized aggregates that adhere to the tumor vasculature only when tumors are heated to 41.5 degrees C. Upon return to normothermia, the vascular particles dissolve into the plasma, increasing the vascular concentration, which drives more ELPs across the tumor blood vessel and significantly increases its extravascular accumulation. These observations suggested that thermal cycling of tumors would increase the exposure of tumor cells to ELP drug carriers. We investigated this hypothesis in this study by thermally cycling an implanted tumor in nude mice from body temperature to 41.5 degrees C thrice within 1.5 h, and showed the repeated formation of adherent microparticles of ELP in the heated tumor vasculature in each thermal cycle. These results suggest that thermal cycling of tumors can be repeated multiple times to further increase the accumulation of a thermally responsive polymeric drug carrier in solid tumors over a single heat-cool cycle. More broadly, this study shows a new approach--tumor thermal cycling--to exploit stimuli-responsive polymers in vivo to target the tumor vasculature or extravascular compartment with high specificity.

Topics: Animals; Carcinoma, Squamous Cell; Combined Modality Therapy; Drug Delivery Systems; Elastin; Humans; Hydrazines; Hyperthermia, Induced; Mice; Mice, Inbred BALB C; Mice, Nude; Peptides; Quinolinium Compounds

Thermally responsive elastin-like polypeptides (ELPs) were synthesized by recombinant DNA techniques and conjugated to doxorubicin through an acid-labile hydrazone bond to enable release of the drug in the acidic environment of lysosomes. The thermal properties, intracellular localization and cytotoxicity of the conjugate were investigated in this study. The conjugation procedure resulted in a mixed population of free ELP and ELP-doxorubicin (ELP-dox) conjugates that exhibit a broader transition than the parent ELP. A simple centrifugation procedure was developed to purify the ELP-dox conjugate from other reactants and resulted in a sharper thermal transition, similar to the parent ELP. The ELP was endocytosed by squamous cell carcinoma cells (FaDu) and trafficked into lysosomes, as observed by the colocalization of the ELP with a lysosome-specific dye through confocal fluorescence microscopy. Interestingly, both the ELP-dox conjugate and free drug exhibited near equivalent in vitro cytotoxicity, although their subcellular localization was significantly different. The free drug was largely concentrated in the nucleus, while the conjugate was dispersed throughout the cytoplasm with limited nuclear accumulation. These differences are significant because they suggest a different mechanism of cytotoxicity for the conjugate as compared with the free drug.

Topics: Antibiotics, Antineoplastic; Carcinoma, Squamous Cell; Cell Survival; Chemical Phenomena; Chemistry, Physical; Doxorubicin; Elastin; Humans; Hydrazones; Microscopy, Confocal; Neoplasms; Peptides; Thermodynamics; Tumor Cells, Cultured

Elastin-like polypeptides (ELPs) composed of a VPGXG repeat undergo a reversible phase transition in aqueous solution. They are hydrophilic and soluble in aqueous solution below their transition temperature (T(t)), but they become hydrophobic and aggregate when the temperature is raised above their T(t). In this study, we examine the quantitative uptake of a fluorescence-labeled, thermally responsive ELP as a function of ELP concentration between 5 and 15 microM in solution in response to hyperthermia by three cultured cancer cell lines. Flow cytometry of fluorescein-ELP conjugates showed that hyperthermia enhanced the cellular uptake of the thermally responsive ELP in human ovarian carcinoma cells (SKOV-3) and in HeLa cells at a concentration of 10 microM or higher, but not at a concentration of 5 microM, as compared with the uptake of a thermally inactive ELP control. In FaDu cells, hyperthermia stimulated uptake of the thermally responsive ELP at all solution concentrations of ELP between 5 and 15 microM. In particular, a >2-fold greater uptake of thermally responsive ELP compared with the thermally inactive control ELP was observed for FaDu cells at a solution concentration of 15 microM in heated cells. Confocal fluorescence microscopy of tumor cells incubated with a rhodamine conjugate of the thermally responsive ELP showed that the cytoplasm was uniformly stained below the T(t). Above the T(t), fluorescent particles were observed in the cytoplasm, suggesting that these particles are aggregates of the thermally responsive polypeptide resulting from the ELP phase transition. These studies demonstrate that the endocytotic uptake of a thermally responsive ELP is significantly enhanced by the thermally triggered phase transition of the polypeptide.

Topics: Carcinoma, Squamous Cell; Drug Carriers; Elastin; Female; Flow Cytometry; Fluorescent Dyes; Hot Temperature; Humans; Hyperthermia, Induced; Kinetics; Microscopy, Confocal; Ovarian Neoplasms; Peptides; Subcellular Fractions; Tumor Cells, Cultured

Elastin metabolism parameters (elastin-derived peptides and elastase-like activity) were determined in sera of patients with lung cancer and in healthy controls. The concentration of elastin-derived peptides was statistically significantly elevated in the lung cancer group. There was no statistically significant difference in the serum elastase-like activity between the groups studied. These data seem to indicate an enhanced metabolism of elastin in patients with lung cancer.

Topics: Adenocarcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Elastin; Female; Humans; Lung Neoplasms; Male; Middle Aged; Pancreatic Elastase; Peptide Fragments

The aim of this study was to investigate anti-elastin antibodies of the IgG and IgM types in sera of patients suffering from lung cancer, using the DOT immunobinding assay. We studied 96 pathological and 40 control sera. Anti-elastin antibodies were found to be present in 45% of patients with small cell lung cancer, 19% of subjects with adenocarcinoma and not-identified lung tumor and 15% of patients with squamous cell lung cancer. They circulated in 5% of control persons only. The highest values of their titers were observed in the advanced stages of disease. In 55% of anti-elastin antibody positive small cell lung cancer patients, antibodies were of the IgM type, suggesting the initial step of the autoimmunization to elastin.

Topics: Adenocarcinoma; Antibodies, Neoplasm; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Elastin; Female; Humans; Immunoblotting; Immunoglobulin G; Immunoglobulin M; Lung Neoplasms; Male; Middle Aged

Topics: Aging; Amino Acids; Carcinoma, Squamous Cell; Collagen; Elastin; Geriatrics; Histology; Humans; Keratosis; Keratosis, Actinic; Radiation Effects; Skin; Skin Aging; Skin Diseases; Skin Neoplasms; Sunlight; Ultraviolet Rays