Growth Factor Expression In Tumors Research Articles

Tumor‑suppressive effects of Smad‑ubiquitination regulator 2 in papillary thyroid carcinoma.

Smad-ubiquitination regulator 2 (SMURF2) functions as a homolog of E6AP carboxyl terminus-type E3 ubiquitin ligase to regulate cell cycle progression and tumor growth factor expression. SMURF2 has been revealed to function as a tumor suppressor in a number of cancers; however, its function in papillary thyroid carcinoma (PTC) remains largely unknown. Therefore, the aim of the present study was to investigate the function of SMURF2 in PTC. Reverse transcription-quantitative PCR and western blotting were used to detect cellular expression of SMURF2 in vitro. After increasing or inhibiting the expression of SMURF2, MTT was used to detect the effect on tumor cell proliferation and Transwell assays were used to detect the effect on tumor cell migration and invasion. Finally, ELISA was used to detect the effects on glucose and glutamine metabolism in tumor cells and the findings revealed that SMURF2 was downregulated in PTC tissues. Moreover, SMURF2 inhibited the proliferation, invasion and migration of PTC cells, and promoted their apoptosis. Finally, SMURF2 inhibited cell glycolysis and glutaminolysis and affected metabolism in the PTC cell line, TPC-1. Thus, the findings of the present study suggest that SMURF2 may be a potential target in the treatment of PTC.

TL1A promotes metastasis and EMT process of colorectal cancer

BackgroundMetastasis is the major problem of colorectal cancer (CRC) and is correlated with the high mortality. Tumor necrosis factor-like cytokine 1A (TL1A) is a novel regulatory factor for inflammatory diseases. This work aimed to investigate the role of TL1A in CRC metastasis. MethodAOM/DSS-induced mouse model, xenograft tumor model and metastasis murine model were established to mimic the colitis-associated CRC and investigate CRC growth and metastasis in vivo. Colon tissues were assessed by hematoxylin/eosin (HE) staining and immunohistochemistry (IHC). CRC cell metastasis in vivo was observed using in vivo imaging system (IVIS). Cell viability and proliferation were examined using cell counting kit 8 (CCK-8) and EdU experiments. The expression of tumor growth factor β (TGFβ) and metastatic biomarkers were detected using western blotting experiment. The in vitro cell metastasis was measured by Transwell. ResultsKnockdown of TL1A notably suppressed the generation of colonic tumors in azoxymethane/dextran sodium sulfate (AOM/DSS) model, suppressed in vivo CRC cell growth, as well as lung and liver metastasis. The inflammation response and inflammatory cell infiltration in tumor sites were decreased by TL1A depletion. The in vitro CRC cell growth and metastasis was also suppressed by shTL1A, along with altered expression of epithelial mesenchymal transition (EMT) biomarkers. TL1A depletion suppressed the level of the TGF-β1 receptor (TβRI) and phosphorylation of Smad3 in CRC cells. Stimulation with TGF-β recovered the CRC cell migration and invasion that suppressed by shTL1A. ConclusionOur work implicated TL1A as a promoter of CRC generation and metastasis and defines TGF-β/Smad3 signaling as mediator of TL1A-regualated CRC cell metastasis.

Modulatory Effects of Estradiol and Its Mixtures with Ligands of GPER and PPAR on MAPK and PI3K/Akt Signaling Pathways and Tumorigenic Factors in Mouse Testis Explants and Mouse Tumor Leydig Cells.

The present study was designed to evaluate how estradiol alone or in combination with G protein-coupled estrogen receptor (GPER) agonists and GPER and peroxisome proliferator-activated receptor (PPAR) antagonists alter the expression of tumor growth factor β (TGF-β), cyclooxygenase-2 (COX-2), hypoxia inducible factor 1-alpha (HIF-1α), and vascular endothelial growth factor (VEGF) in mouse testis explants and MA-10 mouse tumor Leydig cells. In order to define the hormone-associated signaling pathway, the expression of MAPK and PI3K/Akt was also examined. Tissue explants and cells were treated with estradiol as well as GPER agonist (ICI 182,780), GPER antagonist (G-15), PPARα antagonist (GW6471), and PPARγ antagonist (T00709072) in various combinations. First, we showed that in testis explants GPER and PPARα expressions were activated by the GPER agonist and estradiol (either alone or in mixtures), whereas PPARγ expression was activated only by GPER agonist. Second, increased TGF-β expression and decreased COX-2 expression were found in all experimental groups of testicular explants and MA-10 cells, except for up-regulated COX-2 expression in estradiol-treated cells, compared to respective controls. Third, estradiol treatment led to elevated expression of HIF-1α and VEGF, while their lower levels versus control were noted in the remaining groups of explants. Finally, we demonstrated the up-regulation of MAPK and PI3Kp85/Akt expressions in estradiol-treated groups of both ex vivo and in vitro models, whereas estradiol in mixtures with compounds of agonistic or antagonistic properties either up-regulated or down-regulated signaling kinase expression levels. Our results suggest that a balanced estrogen level and its action together with proper GPER and PPAR signaling play a key role in the maintenance of testis homeostasis. Moreover, changes in TGF-β and COX-2 expressions (that disrupted estrogen pathway) as well as disturbed GPER-PPAR signaling observed after estradiol treatment may be involved in testicular tumorigenesis.

Hapten sensitization to vaginal mucosa induces less recruitment of dendritic cells accompanying TGF‐β‐expressing CD206+ cells compared with skin

IntroductionContact hypersensitivity (CHS), a type of delayed‐type hypersensitivity, is induced by hapten exposure to the skin and mucosa. We previously reported that, in a murine model of CHS, the vaginal mucosa (VM) sensitization showed lower T‐cell responses as compared with the abdominal skin sensitization. To investigate mechanisms of impaired CHS by the VM sensitization, we compared migration of hapten‐captured dendritic cells (DCs) in the draining lymph nodes (dLNs) and recruitment of DCs at the sensitized local sites.MethodsFluorescein isothiocyanate (FITC) or 2,4‑dinitrofluorobenzene (DNFB) was used as hapten, and migration of FITC+ DCs in the dLNs and local recruitment of MHC class II+ and CD11c+ cells were compared between abdominal skin and VM sensitization by flow cytometric analyses and immunohistochemistry. Expression of tumor growth factor (TGF)‐β at mRNA and protein levels, and local recruitment of CD206+ cells were examined after VM sensitization.ResultsVM sensitization showed less numbers of FITC+MHC class IIhighCD11c+ migratory DCs in the dLNs at 6 and 24 h, as compared with skin sensitization. Both skin and VM sensitization induced the recruitment of dermal/submucosal DCs at 6 h, but the number of submucosal DCs in the VM was significantly decreased at 24 h. VM showed persistently higher mRNA levels of TGF‐β2/β3 expression than those of the skin before and after sensitization. In the VM sensitization, increment of CD206+MHC class II+ cells was observed especially at the deep lamina propria at 24 h. Most of CD206+ cells were also positive for the binding to Fc chimeric TGF‐β receptor that interacts with all TGF‐β isoforms, suggesting TGF‐β expression.ConclusionDC migration to dLNs and localization of DCs at the sensitized sites are limited in the VM sensitization. Our results suggest that the existence of TGF‐β‐expressing CD206+ cells may contribute less sensitization ability and CHS responses in the VM.

Copper-doped MOF-based nanocomposite for GSH depleted chemo/photothermal/chemodynamic combination therapy

Chemodynamic therapy (CDT) can kill cancer cells by generating toxic agents within tumors through Fenton-/Fenton-like reaction, exhibiting high tumor specificity and low systemic toxicity. However, the therapeutic effect of CDT is usually limited by the overexpressed glutathione (GSH) and low catalytic efficiency in the tumor microenvironment. To deplete GSH and achieve chemo/photothermal/chemodynamic combination therapy, Cu2+, the photothermal material polydopamine (PDA), and chemotherapeutic agent DOX were used to construct a copper-doped MOF-based nanocomposite (DOX@Cu2+/ZIF-8@PDA, DCZP). The Cu2+ could be reduced to Cu+ by GSH to achieve GSH depletion and CDT activation. Under laser irradiation, the heat generated by DCZP could be used for photothermal therapy and promote the Fenton-like reaction with the released DOX to achieve combination therapy. Extracellular experiments demonstrated that more than 50% of GSH could be consumed and the generated heat upon laser irradiation could promote •OH production. In vitro experiments showed that the DCZP combined therapy resulted in 49.4% of late cell apoptosis. More importantly, under the combination therapy of DCZP, the tumors of mice were almost ablated and the expression of tumor growth factors also could be suppressed. The work provided a promising strategy by constructing a GSH depletion and combination therapy nanocomposite for safe and efficient cancer therapy.

Effects of administration of probiotic lactobacilli on immunity conferred by the herpesvirus of turkeys vaccine against challenge with a very virulent Marek’s disease virus in chickens

Several vaccines have been used to control Marek’s disease (MD) in chickens. However, the emergence of new strains of Marek’s disease virus (MDV) imposes a threat to vaccine efficacy. Therefore, the current study was carried out to investigate whether concurrent administration of probiotics with the herpesvirus of turkeys (HVT) vaccine enhances its protective efficacy against MDV infection. In this regard, a cocktail comprised of four Lactobacillus species was administered with HVT to chicken embryos at embryonic day 18 (ED18) and/or from day 1 to day 4 post-hatch. The results revealed that the administration of a probiotic Lactobacillus with HVT at ED18 followed by oral gavage with the same lactobacilli cocktail to newly hatched chicks for the first 4 days post-hatch increased the expression of major histocompatibility complex (MHC) II on macrophages and B cells in spleen and decreased the number of CD4+CD25+ T regulatory cells in the spleen. Subsequently, chicks were infected with MDV. The chickens that received in ovo HVT and lactobacilli or HVT had higher expression of IFN-α at 21dpi in the spleen compared to the chickens that were challenged with MDV. Also, the expression of IFN-β in cecal tonsils at 10dpi was higher in the groups that received in ovo HVT and lactobacilli and oral lactobacilli compared to the group that received in ovo HVT alone. Moreover, the expression of tumor growth factor (TGF)-β4 at 4 days post-infection was reduced in the group that received both HVT and probiotics at ED18. Additionally, concurrent probiotics administration reduced tumor incidence by half when compared to HVT vaccine alone indicating enhancing effect of lactobacilli with HVT vaccine on host immune responses. In conclusion, these findings suggest the potential use of probiotic lactobacilli as adjuvants with the HVT vaccine against MDV infection in chickens.

Abstract B56: RNA-seq analysis of prostate cancer samples from Hispanic patients reveal progressive characteristics in Gleason patterns

Abstract Background. One of the prognostic markers widely used in clinic to predict survival in prostate cancer (PC) patients is the Gleason score. Differences have been observed in biochemical progression-free and overall survival in patients with different Gleason scores. Because each Gleason represents a different state of tumor progression, we decided to characterize expression profiles and pathways in Gleason 3, 4 and 5 in order to explore the tumor biology context in each pattern, and thus determine possible prognosis biomarkers in PC. Methods. We extracted RNA from normal and malignant formalin-fixed paraffin-embedded tissues from radical prostatectomy samples of Colombian patients and analyzed expression using data generated with next generation sequencing methods. Pathway analysis was done with MetaCore. Results. Eleven gene networks (7 genes) were common between all Gleason scores and represented cellular processes possibly involved with damage of the prostatic tissue including: cell adhesion, cytoskeleton remodeling and metabolism of phosphatidyl compounds. According to the analysis in Gleason pattern 3, the principal pathways were associated with cytoskeleton remodeling, gap junctions signaling and regulation of cell adhesion. These pathways could contribute to de-differentiation process experienced by cells towards tumor transformation in early stages of progression. Also, signaling of epithelial-to-mesenchymal transition (EMT) were observed, this findings could suggests not only cytoskeleton remodeling during de-differentiation, but also appearance of metastatic cells in this Gleason pattern. On the other hand, the action of the Androgen Receptor (AR) is suggested by protein-protein interactions characterized by rapid activation of signaling cascades of proliferation, cell growth and survival, via MAPK and PI3K/AKT. The latter in turn induces AR overexpression through control of regulatory protein MDM-2 and optimizes binding of AR to promoter/enhancer regions over its target genes. Also, AR inhibits WNT signaling via interaction with B-catenin. However, AR is a LEF-1/TCF transcriptional target, which is activated by WNT/B-catenin signaling. The pathways identified in Gleason pattern 4 were similar to those of Gleason 3, with a more significant participation of pathways leading to cytoskeleton remodeling, regulation of cell adhesion molecules and of EMT. In addition, there is ligand-independent activation of AR, wherein activating mutations, splicing variants and activation by other signaling cascades, may lead to directed transactivation of target genes in a deregulated manner. Further, NGF/TrkA/PI3K-mediated signaling may lead to inhibition of apoptosis and survival of these cells. Finally, pathways analysis in Gleason pattern 5 showed that the most significant was cell adhesion and ECM remodeling, which favors the metastatic process and regulation of cell motility and adhesion, processes aided by migration and angiogenesis signaling pathways. Likewise, AR activation and downstream signaling has an important role, transactivating transcription factors, which together lead to cell proliferation, inhibition of apoptosis, expression of tumor growth factors, regulation of cell adhesion molecules and from inflammatory response. The latter is reflected in another pathway identified, as is the antigen presentation by MHC class II. Conclusion. The pathways identified by different Gleason patterns exhibit progression of localized prostate tumors into metastatic ones, where there is cell growth and dysregulation in signaling pathways at the beginning, and then, as they progress, they acquire the ability to migrate and to regulate AR signaling through different mechanisms so that allows AR to be a central regulator in PC. Citation Format: Natalia L. Acosta, Melody C. Baddoo, Alba L. Combita, Rodolfo Varela, Jorge Mesa, Maria C. Sanabria-Salas, Jovanny Zabaleta. RNA-seq analysis of prostate cancer samples from Hispanic patients reveal progressive characteristics in Gleason patterns. [abstract]. In: Proceedings of the Ninth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2016 Sep 25-28; Fort Lauderdale, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(2 Suppl):Abstract nr B56.

Bisphenol-A induces expression of HOXC6, an estrogen-regulated homeobox-containing gene associated with breast cancer

HOXC6 is a homeobox-containing gene associated with mammary gland development and is overexpressed in variety of cancers including breast and prostate cancers. Here, we have examined the expression of HOXC6 in breast cancer tissue, investigated its transcriptional regulation via estradiol (E2) and bisphenol-A (BPA, an estrogenic endocrine disruptor) in vitro and in vivo. We observed that HOXC6 is differentially over-expressed in breast cancer tissue. E2 induces HOXC6 expression in cultured breast cancer cells and in mammary glands of Sprague Dawley rats. HOXC6 expression is also induced upon exposure to BPA both in vitro and in vivo. Estrogen-receptor-alpha (ERα) and ER-coregulators such as MLL-histone methylases are bound to the HOXC6 promoter upon exposure to E2 or BPA and that resulted in increased histone H3K4-trimethylation, histone acetylation, and recruitment of RNA polymerase II at the HOXC6 promoter. HOXC6 overexpression induces expression of tumor growth factors and facilitates growth 3D-colony formation, indicating its potential roles in tumor growth. Our studies demonstrate that HOXC6, which is a critical player in mammary gland development, is upregulated in multiple cases of breast cancer, and is transcriptionally regulated by E2 and BPA, in vitro and in vivo.

B-cell translocation 1 gene inhibits cellular metastasis-associated behavior in breast cancer

B-cell translocation gene 1 (BTG1) is a member of the BTG/transducer of ERBB2 family, which regulates cell cycle progression in a variety of cell types and may have a role in inhibiting proliferation, promoting apoptosis and stimulating cellular differentiation in numerous cell types. However, the role of BTG1 in cancer metastasis is yet to be elucidated. In the present study, analysis of clinical specimens revealed that BTG1 mRNA levels were lower in lymph node metastases than those in benign breast tumors and normal human breast tissue. The effect of BTG1 on the metastatic behavior of breast cancer cells following stable transfection with a BTG1 expression vector was also investigated. The overexpression of BTG1 was observed to inhibit cell adhesion, migration and invasion. Furthermore, the overexpression of BTG1 was found to be involved in the inhibition of the metastasis-related proteins matrix metalloproteinase-2 and -9, as well as the promotion of the cell-cell adhesion-associated protein E-cadherin. In syngeneic nude mice breast tumor models, hepatic metastasis and angiogenesis were observed in the mice injected with the control cells, but not in those injected with pcDNA3-BTG1 cells. Immunohistochemistry revealed that overexpression of BTG1 decreased vascular endothelial growth factor expression in tumors. To the best of our knowledge, this is the first study to show that BTG1 overexpression decreases migration and invasion of breast cancer cells and thereby inhibits distant metastasis in mice breast tumor models.

Malignant Ovarian Tumors with Induced Expression of Carbonyl Reductase Show Spontaneous Regression

BackgroundThe present study investigated tumor proliferation in a tumor model using murine ovarian cancer cells with increased carbonyl reductase (CR) expression.MethodsCR cDNA was transfected into murine T-Ag-MOSE ovarian cancer cells by lipofection. CR-transfected cells (CR induction group) or empty vector-treated cells (control group) were injected into the backs of 8-week-old nude mice at a concentration of 0.5 × 106 per 0.2 mL. Subsequent tumor proliferation in both groups was observed for 5 weeks.ResultsThe control group showed an increase in tumor volume during the 5 weeks of observation. However, tumor volume in the CR induction group increased up to the second week but then decreased continuously until the fifth week of observation. The tumor growth curves for the two groups showed a significant difference (Mann-Whitney U test, P < 0.001). Histological and biochemical experiments were performed using tumor tissues isolated in the third week. Necrosis and inflammatory cell infiltration were noted for tumors in the CR induction group. Also, the number of apoptotic cells was significantly increased in the CR induction group compared with the control group (P < 0.001). Milk fat globule EGF factor 8, an “eat-me” signal for phagocytes such as macrophages, was expressed extensively in the tumor cytoplasm and interstitial cells of the CR induction group, and engulfment of apoptotic cells by macrophages was observed. Vascular endothelial growth factor expression in tumors was notably decreased in the CR induction group compared with the control group.ConclusionIncreased necrosis due to engulfing of apoptotic cells by phagocytes attracted by increased milk fat globule EGF factor 8 was considered to be the mechanism of spontaneous tumor regression in the CR induction group.

Tumor angiogenesis in predicting the survival of patients with stage I lung cancer

The effects of angiogenesis on survival were assessed by measuring the tumor microvessel density and vascular endothelial growth factor expression in patients with resected stage I non-small cell lung carcinoma. The study population included 141 patients who underwent complete resection for stage pT1 and T2 N0 M0 tumors between 1999 and 2007. Lobectomy and pneumonectomy were performed in 131 and 10 patients, respectively. Tumor specimens were analyzed immunohistochemically for staining with anti-CD105 antibody to determine tumor microvessel density and anti-vascular endothelial growth factor antibody to determine the vascular endothelial growth factor expression level. Univariate and multivariate analyses were performed for factors influencing patients' survival. The overall 5-year survival was calculated as 68%, with rates of 76.9% for patients with T1 disease and 66.2% for patients with T2 disease (P=.4). The vascular endothelial growth factor expression rate was 94.3% for patients with stage I non-small cell lung carcinoma. Vascular endothelial growth factor expression did not influence survival (P=.9). The median microvessel density of the tumors measured based on the level of CD105 expression was 19.8. The effect of microvessel density on survival was significant (P=.02). The 5-year survivals of patients with tumors with 20 or more microvessels and less than 20 microvessels were 76.8% and 56.1%, respectively; this difference was highly significant (P=.004). The microvessel density was determined as an independent factor influencing survival on multivariate analysis (P=.03). The level of vascular endothelial growth factor expression in tumors was not a successful predictor of survival in patients with resected stage I non-small cell lung carcinoma. A high microvessel density based on CD105 is a strong predictor of prognosis in these patients.

Tumor growth factor expression in obesity and changes in expression with weight loss: another cause of increased virulence and incidence of cancer in obesity

Background Obesity is associated with increased tumerogenesis. Previously, we demonstrated that inflammation in obesity caused cancer fighting cells to display greater surface receptor levels, predisposing them to early cell death. We measured the inflammatory tumor growth factor levels to determine whether inflammation in obesity increases expression of these factors, potentially predisposing these patients to greater rates of neoplasia. Methods A total of 24 patients undergoing weight loss surgery had samples collected preoperatively and at 6 and 12 months after surgery. The growth factors analyzed included tumor necrosis factor (TNF)-α, granulocyte-macrophage colony-stimulating factor, interferon-γ, interleukin (IL)-1b, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, vascular endothelial growth factor, hepatocyte growth factor, TNF-receptor I (TNF-RI), TNF-RII, death receptor 5, leptin, and adiponectin. Control samples were obtained from 10 healthy, normal weight volunteers. Results The tumor growth factors TNF-α, TNF-RI, TNF-RII, vascular endothelial growth factor, hepatocyte growth factor, interferon-γ, IL-2, IL-5, and IL-6 all decreased significantly ( P <.05) compared with the preoperative values. The IL-4, IL-8, leptin, death receptor 5, adiponectin, and granulocyte-macrophage colony-stimulating factor levels did not change significantly over time. The IL-1b and IL-10 levels were less than the detection limit at all points. When obese patient serum was compared with healthy volunteer pooled serum, we found that the leptin, death receptor 5, hepatocyte growth factor, vascular endothelial growth factor, TNF-RI, TNF-RII, TNF-α, IFN-γ, granulocyte-macrophage colony-stimulating factor, IL-4, IL-5, IL-6, and IL-8 levels were all 2–37 times greater than the levels in the controls at baseline. The concentrations of these same growth factors had decreased levels only 1–3.5 times greater than those of the controls at 12 months postoperatively. Conclusion Many inflammatory tumor growth factors are present in greater concentrations in obese individuals. This could explain the greater prevalence of neoplasia in the morbidly obese population.

PL-305: The effect of weight loss surgery on the expression of tumor growth factors in morbid obesity

Obesity is associated with increased tumerogenesis. Previously,we demonstrated that inflammation in obesity caused cancer fighting cells to display higher surface receptors levels predisposing them to early cell death. Herein, we measured inflammatory tumor growth factors to determine whether inflammation in obesity increases expression of these factors potentially predisposing these patients to higher rates of neoplasia.

Summary: Tissue Remodeling and Repair Mechanisms in Chronic Obstructive Pulmonary Disease

Nearly all drugs used for treatment of airway disease are homologs or close derivates of the sympathomimetics, parasympatholytics, cromones, andmethylxanthines that were developedmore than 50 years ago. By the standards of classical reversible airflow obstruction (e.g., asthma), the application of these derivatives to the treatment of chronic obstructive pulmonary disease (COPD) has been marginally successful in reversing airflow obstruction and relieving symptoms. The reason for this is the nature of the anatomic changes in COPD. Bronchodilation is a poor target for treatment of COPD if the goal is to restore patients with advanced disease to near-normal lung function. This conference was convened to examine the possibilities for the distant future (20 years or more)—possibilities that will lead to the restoration of normal lung structure in lungs that are severely damaged from COPD. The conference was entitled “Tissue Remodeling and Repair in COPD.” By repair, the conferees were examining the hypothesis that an understanding of the process by which lung development occurs could be harnessed to redevelop new, normal lung units where terminal damage had already occurred. Much of the conference focused on alveolation, and, hence, on classically defined emphysema only. COPD is nonetheless an airway and alveolar disease, and it is not clear that the same processes would apply for repair of conducting airways and alveoli. As summarized by David Warburton, lung development starts at 5 weeks, at which time a tracheal evagination from the esophagus can be visualized. The conducting portion of the airway extends to branch ultimately into alveolated tissue. The conducting airways at each branch point are sacrificed for the next generation so that the 300 million alveoli from which we exchange gas are formed from a single ancestral bud. Airway branching is mostly complete at birth, but the development of alveolation continues to age 8 years. Several important points emerged from this discussion. First, airways in the fetus and young infant develop in an optimal environment over a normal substrate that is not damaged by disease. Second, timing is everything. The brilliant choreography of lung branching and morphogenesis is precisely timed. Specific genes have been identified, but generating new lung from proximal airways requires timely promotion and termination of gene expression. The developmental priority lies in the stem cell hierarchy. Improper expression of the FgF-10 gene can lead to massive overalveolation. Vascular endothelial growth factor, which controls vascular morphogenesis when overexpressed in mice, leads to devastating hypervascularity. Specific mutations that affect elastin expression can create an emphysema-like phenotype. Excessive expression of tumor growth factor (TGF)is particularly enigmatic. Overexpression

Hepatocyte growth factor ameliorates dermal sclerosis in the tight-skin mouse model of scleroderma

The tight-skin (TSK/+) mouse, a genetic model of systemic sclerosis (SSc), develops cutaneous fibrosis and defects in pulmonary architecture. Because hepatocyte growth factor (HGF) is an important mitogen and morphogen that contributes to the repair process after tissue injury, we investigated the role of HGF in cutaneous fibrosis and pulmonary architecture defects in SSc using TSK/+ mice. TSK/+ mice were injected in the gluteal muscle with either hemagglutinating virus of Japan (HVJ) liposomes containing 8 μg of a human HGF expression vector (HGF-HVJ liposomes) or a mock vector (untreated control). Gene transfer was repeated once weekly for 8 weeks. The effects of HGF gene transfection on the histopathology and expression of tumor growth factor (TGF)-β and IL-4 mRNA in TSK/+ mice were examined. The effect of recombinant HGF on IL-4 production by TSK/+ CD4+ T cells stimulated by allogeneic dendritic cells (DCs) in vitro was also examined. Histologic analysis revealed that HGF gene transfection in TSK/+ mice resulted in a marked reduction of hypodermal thickness, including the subcutaneous connective tissue layer. The hypodermal thickness of HGF-treated TSK/+ mice was decreased two-fold to three-fold compared with untreated TSK/+ mice. However, TSK/+ associated defects in pulmonary architecture were unaffected by HGF gene transfection. HGF gene transfection significantly inhibited the expression of IL-4 and TGF-β1 mRNA in the spleen and skin but not in the lung. We also performed a mixed lymphocyte culture and examined the effect of recombinant HGF on the generation of IL-4. Recombinant HGF significantly inhibited IL-4 production in TSK/+ CD4+ T cells stimulated by allogeneic DCs. HGF gene transfection inhibited IL-4 and TGF-β mRNA expression, which has been postulated to have a major role in fibrinogenesis and reduced hypodermal thickness, including the subcutaneous connective tissue layer of TSK/+ mice. HGF might represent a novel strategy for the treatment of SSc.

Effect of astrgaloside IV on the long-term consequences of renal ischemia-reperfusion injury in rat

To observe the effect of astrgaloside IV (Astr) on the long-term consequences of renal ischemia-reperfusion injury (IRI) in rat. Fifty-four male Sprague-Dawley rats were randomized into 3 equal groups: IRI group, Astr group, and sham operation group. All rats underwent right nephrectomy and isolation of the left renal artery. The left renal arteries of the IRI group and Astr group were gripped by vascular clamp for 60 minutes and that of the sham operation group was only isolated without gripping. Two milliliters of Astr solution (0.1 g/L) was perfused into the stomach of the rats in the Astr group three days before and after the operation respectively. The rats in the IRI and sham operation groups were perfused with normal saline of the same volume. Four, twelve, and twenty-four weeks after the operation 24-hour urine specimens of the rats were collected to detect the urine protein. At each time point 6 rats from each group were anesthetized and blood was collected from the abdominal aorta to measure the level of serum creatinine (Cr), their left kidneys were taken out to undergo pathological examination and extraction of mRNA. Histochemistry was used to detect the expression of tumor growth factor (TGF)-beta1 protein in the renal tissues. RT-PCR was used to detect the expression of TGF-beta1 mRNA. Collagen staining and immunohistochemistry were used to measure the proportion of collagen positive material to the total area. The level of urine protein was increased progressively, those 12 and 24 weeks after the operation in the IRI group were significantly higher than those in the Astr and sham operation groups (all P < 0.05). The serum Cr 4 weeks after the operation was 36 micromol/L +/- 4 micromol/L, significantly higher than those in the Astr and sham operation groups (31 micromol/L +/- 8 micromol/L and 31 micromol/L +/- 5 micromol/L), and the serum Cr levels 4 weeks 12 and 24 weeks after the operation in the IRI group remained significantly higher than those in the Astr and sham operation groups (all P < 0.05). Collagen staining showed that the glomerular basement membrane, tunica adventitia vasorum, and adventitia of renal tubule were remarkably redder in the IRI than in the Astr and sham operation groups. The expression of TGF-beta1 protein was progressively increased since 12 weeks after the operation in the IRI group, significantly stronger in the Astr and sham operation groups. The expression of TGF-beta1 mRNA was progressively increased since 12 weeks after the operation in the IRI and Astr groups, significantly stronger than that in the sham operation group (P < 0.05). However, the expression of TGF-beta1 mRNA 24 weeks after the operation was significantly stronger in the IRI group than in the Astr group (P < 0.05). After renal IRI the probability of development of renal fibrosis increases. Astrgaloside IV markedly ameliorates renal injury by downregulating the TGF-beta1 expression.

Do Angiogenesis and Growth Factor Expression Predict Prognosis of Esophageal Cancer?

A retrospective study of surgically resectable esophageal cancers was undertaken to determine the relationship between angiogenesis score and growth factor expression with tumor size, histology, degree of differentiation, depth of invasion, nodal disease, and the presence of Barrett's esophagus. The office and hospital charts of 27 patients who had esophageal resection for carcinoma between 1990 and 1995 at Rush-Presbyterian-St. Luke's Medical Center were reviewed. Data collection included patient demographics, survival, tumor size, histology, differentiation, depth of invasion, nodal metastases, and the presence of Barrett's esophagus. The pathology specimens were immunostained for von Willebrand factor (factor VIII-related antigen). Immunostaining was also performed for vascular endothelial growth factor and transforming growth factor alpha. Twenty normal esophageal specimens served as controls. Angiogenesis score was determined by counting vessels under conventional light microscopy at x200 magnification, and growth factor expression was graded on a scale of 1 to 4. Cancers had higher angiogenesis and growth factor expression than controls (P = 0.01). Patient age, tumor size, histology, differentiation, depth of invasion, and Barrett's esophagus did not correlate with angiogenesis score or tumor growth factor expression. Lymph node status did correlate with both angiogenesis score and growth factor expression (P < or = 0.02). We conclude that high angiogenesis score and growth factor expression correlate with the presence of lymph node metastases. This may help select patients for preoperative radiation and chemotherapy or determine the extent of surgery performed for esophageal carcinoma.