An Autopsy Case of Primary Gastric Choriocarcinoma

An autopsy case of primary gastric choriocarcinoma in a 55-year-oldmale is presented. The tumor was diagnosed as choriocarcinomaof the stomach from a histological examination of biopsy specimens.The level of human chorionic gonadotropin (HCG) was significantlyincreased in the serum and urine. A histological examinationof autopsy specimens showed the tumor of the stomach to be apure choriocarcinoma composed of syncytiotrophoblasts and cytotrophoblastswith a mixture of eoslnophilic necrotic tissues, but with noelements of adenocarcinoma. The tumor showed metastases and/orinvasions to the liver, lungs, pancreas, omentum, pleura, peritoneumand lymph nodes. Positive immuno-histochemical staining forthe ß-subunit of HCG (HCG-ß) in the gastrictumor was demonstrated in the form of granular diffuse depositsin the cytoplasm of trophoblasts and predominated in syncytiotrophoblastsover transitional cells. Under electron microscopic observation,positive immunostalnlng for HCG-ß was observed inthe perinuclear space, cistemae of the rough endoplasmic reticulum(RER) and secretory vesicles of syncytiotrophoblasts and transitionalcells.     

Cathepsin B immunohistochemical staining in tumor and endothelial cells is a new prognostic factor for survival in patients with brain tumors.

The cysteine endopeptidase, cathepsin (Cat) B, and its endogenous inhibitor, stefin A, were found relevant for cancer progression of many neoplasms, including human brain tumors. Histological sections of 100 primary brain tumors, 27 benign and 73 malignant, were stained immunohistochemically for Cat B and stefin A. The immunohistochemical staining of Cat B in tumor cells, endothelial cells, and macrophages was scored separately from 0-12. The score in tumor and endothelial cells was significantly higher in malignant tumors compared with benign tumors (P<0.000). A significant correlation between immunostaining of Cat B (scored together for tumor and endothelial cells) and clinical parameters, such as duration of symptoms, Karnofsky score, psycho-organic symptoms, and histological score was demonstrated. Univariate survival analysis indicated that total Cat B score above 8 was a significant predictor for shorter overall survival (P = 0.003). In glioblastoma multiforme, intense Cat B staining of endothelial cells was a significant predictor for shorter survival (P = 0.003). Stefin A immunostaining was weak and detected only in a few benign and some malignant tumors, suggesting that this inhibitor alone is not sufficient in balancing proteolytic activity of Cat B. We conclude that specific immunostaining of Cat B in tumor and endothelial cells can be used to predict the risk of death in patients with primary tumors of the central nervous system.     

Enhancement of R3230AC rat mammary tumor growth and cellular differentiation by dibutyryl cyclic adenosine monophosphate.

Daily sc injections of 8 mg N6, O2'-dibutyryl 3',5'-cyclic AMP (DBcAMP) beginning 1 day after tumor implantation significantly increased the growth rate of R32230AC rat mammary adenocarcinomas, which nearly doubled in in situ volume by day 40 compared to similarly implanted tumors in saline-injected controls. Weights of excised tumors, intact, drained, and dried all increased approximately 80%, which suggested that the increase in tumor size was not due to accumulation of secreted fluid or tissue water. Injections of 17beta-estradiol valerate (0.1 mg/wk) from day 1 or of DBcAMP from day 22 resulted in insignificant changes in growth--28% and 35% increases in tumor volume and a 5% decrease and an 18% increase, respectively, in drained wet weight. Electron microscopic examination revealed that estrogen and DBcAMP caused differentiation of the tumor cells into two different states: Estrogen-treated tumors resembled lactating mammary glands; they contained large lipid droplets, organized rough endoplasmic reticulum, and vesicles containing electron dense granules resembling protein. DBcAMP-treated tumor cells were marked by a proliferation of the Golgi complex and numerous vesicles containing fine granular material.     

Turning tumor cells in situ into T-helper cell-stimulating, MHC class II tumor epitope-presenters: immuno-curing and immuno-consolidation

Immunological control or cure of tumors depends on initiating a robust T helper cell response to MHC class II epitopes of tumor-associated antigens. T helper cells regulate the potency of cytotoxic T lymphocyte and antibody responses. We have developed a novel approach to stimulate T helper cells by converting tumor cells into MHC class II molecule-positive, antigen presenting cells. Furthermore, using antisense methods, we suppress expression of the Ii protein, that normally blocks the antigenic peptide binding site of MHC class II molecules during synthesis in the endoplasmic reticulum. In such gene-engineered tumor cells, the MHC class II molecules pick up antigenic peptides, which have been transported into the endoplasmic reticulum for binding to MHC class I molecules. All nucleated cells create such "surveys of self" to detect viral or malignant transformation. Our method extends that survey of self to MHC class II endogenous tumor-associated antigens. Simultaneous presentation of tumor antigens by both MHC class I and II generates a robust and long-lasting antitumor immune response. Injecting murine tumors with genes, which induce MHC class II molecules and suppress Ii protein, cures a significant number of animals with renal and prostate tumors. We have developed analogous human gene vectors that are suitable for most patients and cancers, because they are monomorphic and active in all HLA-DR alleles. We review our findings, and analyze remaining issues for preclinical study and the design of clinical trials.     

The fine structure of astroblastoma

An astroblastoma in the cerebrum was investigated with the electron microscope. A distinctive feature of the tumor was the radial arrangement of astrocytic cells around the blood vessels. The blood vessels in the tumor commonly exhibited fenestrated endothelial cells. The endothelium was generally surrounded by lamellated basal laminae that were compactly invested in the neoplastic cells. The most remarkable feature of the tumor cells was the frequent presence of coated vesicles. The cytoplasm of some tumor cells was filled by delicate glial fibrils and microtubules, whereas that of others contained a moderate amount of rough-surfaced endoplasmic reticulum, free ribosomes, and mitochondria, but only scanty glial fibrils and microtubules. The former usually possessed long, thin cell processes, whereas the latter had poor development of cell processes and commonly showed multiple or giant nuclei containing several nuclear bodies and prominent nucleoli. In summary, this ultrastructural study of astroblastoma revealed the coexistence of varied maturity astrocytic cells and their compact arrangement around fenestrated blood vessels.     

Morphologic and biochemical characterization of erythrophoromas in goldfish (Carassius auratus).

The properties of spontaneous cutaneous erythrophoromas that occurred in 17 adult goldfish (Carassius auratus) were described. This study was based on combined histology, cytochemistry, electron microscopy, and biochemical analysis. Tumors that varied from 0.4 to 2.7 cm in maximum diameter were located on various parts of the skin. As shown by light microscopy, tumors were composed of red- to orange-pigmented, dendritic, stellate, or spindle-shaped cells, which coalesced into an excrescence in the dermal layer. No metastasis was observed. Evidence of invasiveness in five examples and local recurrence in three examples indicated that some of these tumors were malignant. Electron microscopy revealed numerous round cytoplasmic organelles (pterinosomes), which measured 0.4--0.7 mu, and well-developed tubular, smooth endoplasmic reticulum in the tumor cells. Biochemical analysis showed that red pigmentation of tumor cells was imparted by pteridines and carotenoids, most of which were detectable in normal adult erythrophores. These data indicate that tumors were comprised of neoplastic erythrophores.     

细胞内氯离子通道蛋白4与恶性肿瘤发生发展的研究进展

细胞内氯离子通道蛋白4(chloride intracellular channel 4,CLIC4)是细胞内氯离子通道家族成员之一,广泛分布于细胞内,定位于多种细胞器如线粒体、内质网及细胞核和细胞质中,在多种肿瘤组织中表达异常,与肿瘤细胞的黏附、分化、凋亡等生物学行为密切相关,研究其在肿瘤发生、发展过程中的具体作用,将为多种肿瘤的诊断、治疗及预后提供新的依据。     

Interaction of retinoic acid and 3-methylcholanthrene on the fine structure of mouse prostate epithelium in vitro.

The effects of 3-methylcholanthrene (MCA) and retinoic acid (RA) on the fine structure of AKR mouse prostate epithelium in organ culture were correlated with changes in cell proliferation. In intact glands before explantation, the epithelial cytoplasm showed concentric flat or globular cisternae of endoplasmic reticulum in both supranuclear and basal areas, a well-developed Golgi complex, secretory vesicles, and numerous microvilli at the luminal surface. After explantation, the cytoplasmic organelles, particularly the endoplasmic reticulum, regressed and tonofilaments appeared. The regression was largely prevented by RA. MCA induced considerable epithelial hyperplasia and squamous metaplasia. The fine structure of the newly formed cells revealed a complete loss of endoplasmic reticulum, Golgi apparatus, secretory vesicles, and microvilli, with the appearance of bundles of tonofilaments and a striking increase in the number of desmosomes. Administration of RA to explants pretreated with the carcinogen partially reversed the hyperplasia and squamous metaplasia. The tonofilaments disappeared and the number of desmosomes greatly decreased, whereas endoplasmic reticulum, Golgi complex, secretory vesicles, and microvilli were largely reestablished. Planimetric measurements of the alveolar epithelium showed that the squamous transformation and its partial reversal by RA coincide with the rise and decline of epithelial hyperplasia. The data suggest that the restoration of secretory differentiation by RA was responsible for the initial breakdown of the hyperplastic epithelium, whereas the lowering of DNA synthesis by RA prevented further hyperplasia and kept cell replication within normal limits.     

Pleomorphic fibrosarcoma

263 tumors diagnosed as malignant fibrous histiocytomas and undifferentiated pleomorphic sarcomas have been reassessed morphologically, immunohistochemically and ultrastructurally. The diagnosis of pleomorphic fibrosarcoma was made in 19 patients (5%). The tumors occur in the lower and upper extremities and trunk. The patients have a long history of painless intramuscular mass (for a median of 6 months). Follow up showed disease recurrence in 24% of patients and metastases in 32% of patients. All tumors consisted of spindle-shaped fibroblasts and myofibroblasts with a prominent rough endoplasmic reticulum and Golgi apparatus, extracellular collagen fibrils. Variable number of histiocite-like fibroblasts and undifferentiated cells were found. No other signs of tumor cell line differentiation were noted. A pleomorphic fibrosarcoma and malignant fibrous histiocytoma have similar clinical and radiographic manifestation and survival rates.     

Expression of the class VI intermediate filament nestin in human central nervous system tumors.

Tumor cells of a particular tissue may show a pattern of gene expression characteristic of the precursor cells of this tissue. To test this proposition for tumors of the central nervous system (CNS) we have used immunohistochemistry to analyze the expression of nestin in primary human CNS tumors and corresponding nonneoplastic brain tissue. Nestin defines a recently discovered sixth class of intermediate filament proteins and in the rat is expressed predominantly in CNS stem cells. In the adult nonneoplastic human brain we have detected only nestin expression in occasional endothelial cells. In contrast, a variety of primary CNS tumors contained substantially elevated nestin levels. The nestin-positive cells in the tumor tissue were tumor cells and/or endothelial cells. Glioblastomas expressed higher nestin levels than less malignant gliomas. This may indicate a correlation between nestin expression and malignancy within the glioma tumor group. In the primitive neuroectodermal class of tumors we observed both nestin-expressing and nonexpressing tumors, suggesting that nestin expression could be used to further characterize this complex and heterogeneous tumor group. Nine metastatic carcinomas were studied, and none showed nestin immunoreactivity in tumor cells. In conclusion, our data support the notion that primary CNS tumors share gene expression patterns with primitive, undifferentiated CNS cells and that nestin, like other intermediate filaments, may be useful in tumor diagnosis.     

Focal adhesion kinase is expressed in the angiogenic blood vessels of malignant astrocytic tumors in vivo and promotes capillary tube formation of brain microvascular endothelial cells.

PURPOSE: Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase that has been shown to promote proliferation, migration, and invasion of several cell types in vitro, and we have shown recently that FAK promotes proliferation of malignant astrocytoma cells in vivo. To determine the role of FAK in angiogenesis in malignant astrocytic tumors, we investigated the expression and function of FAK in brain endothelial cells. EXPERIMENTAL DESIGN: We characterized the expression of FAK and activated FAK in endothelial cells by immunohistochemistry. We also determined the function of FAK in brain microvascular endothelial cells by transfecting these cells with a dominant interfering form of FAK [FAK-related nonkinase (FRNK)] or a mutant FRNK (Leu-1034 to Ser) and assessed the effect on capillary tube formation and cell migration. RESULTS: We found that FAK was expressed in the endothelial cells of grade III (4 of 9 samples) and IV (9 of 10 samples) astrocytoma biopsies but not in the endothelial cells of normal brain (0 of 9 samples) and not in grade I (0 of 5 samples) or II (0 of 4 samples) astrocytoma biopsies. Furthermore, we found that both FAK and activated FAK were expressed in the endothelial cells in malignant astrocytoma tumors propagated intracerebrally in the severe combined immunodeficient mouse brain. As expected, immunofluorescence analysis showed FRNK protein to localize to focal adhesions, whereas mutant FRNK protein did not. FRNK-transfected endothelial cells showed a 55% reduction in branched tube formation and a 40% reduction in tube length when propagated in three-dimensional collagen gels, compared with cells transfected with the mutant FRNK construct. Furthermore, FRNK-transfected cells showed a 35-50% reduction in haptotactic migration toward fibronectin and collagen, compared with mutant FRNK-transfected cells. CONCLUSIONS: These data suggest that FAK promotes angiogenesis and that this occurs, at least in part, through the promotion of endothelial cell migration.     

The roles of chemokine CXCL12 in embryonic and brain tumor angiogenesis

The formation of blood vessels in embryos and tumors are different processes but under the control of common molecular mechanisms. Chemokine CXCL12 involved in both embryonic and tumor angiogenesis. In this review, we summarize recent advances in understanding the roles of CXCL12 in brain tumor angiogenesis/vasculogenesis. CXCL12 and its cognate receptors are abnormally induced in brain tumors, in particular in tumor cells and endothelium. Pathologically enhanced CXCL12 signaling may promote the formation of new vessels through recruiting circulating endothelial progenitor cells or directly enhancing the migration/growth of endothelial cells. Therefore, CXCL12 signaling represents an important mechanism that regulates brain tumor angiogenesis/vasculogenesis and may provide potential targets for anti-angiogenic therapy in malignant gliomas.     

Efficient inhibition of in vivo human malignant glioma growth and angiogenesis by interferon-beta treatment at early stage of tumor development.

Malignant gliomas are highly angiogenic and aggressive tumors. IFN-beta has been used for the treatment of patients with malignant glioma; however, its antitumor mechanism in vivo remains unclear. To understand the in vivo antitumor effect and mechanism of recombinant human IFN-beta (rhIFN-beta) depending on the stages of tumor development or progression, we used orthotopic xenograft brain tumors generated by stereotactic intracerebral implantation of U-87 human glioma cells in nude mice. Mice bearing tumors 7 days (group 1) and 21 days (group 2) postimplant were treated with 2 x 10(5) IU/day of rhIFN-beta or saline i.p. for 15 days, respectively. Tumor growth was suppressed by 69.6% in group 1 and 10.8% in group 2 compared with tumors of each control group treated with saline. rhIFN-beta-treated group 1 animals showed 38% reduction in vascularization along with a 2.5-fold increase of the apoptotic index and no change in the proliferative index as compared with untreated tumors. The expression level of vascular endothelial cell growth factor and basic fibroblast growth factor was not affected by rhIFN-beta treatment. rhIFN-beta showed inhibitory activity on proliferation of U-87 cells, human umbilical vein endothelial cells, and PAM 212 murine keratinocytes in vitro. Our results indicate that the in vivo antitumor effect of rhIFN-beta on malignant gliomas may be mediated, at least in part, via angiogenesis inhibition rather than antiproliferative activity and that rhIFN-beta may be more effective for the treatment of malignant glioma patients at an early stage with minimal or microscopic tumor burdens rather than at an advanced stage of tumor development.     

SEL1L SNP rs12435998, a predictor of glioblastoma survival and response to radio-chemotherapy

The suppressor of Lin-12-like (C. elegans) (SEL1L) is involved in the endoplasmic reticulum (ER)-associated degradation pathway, malignant transformation and stem cells.In 412 formalin-fixed and paraffin-embedded brain tumors and 39 Glioblastoma multiforme (GBM) cell lines, we determined the frequency of five SEL1L single nucleotide genetic variants with regulatory and coding functions by a SNaPShot™ assay. We tested their possible association with brain tumor risk, prognosis and therapy.We studied the in vitro cytotoxicity of valproic acid (VPA), temozolomide (TMZ), doxorubicin (DOX) and paclitaxel (PTX), alone or in combination, on 11 GBM cell lines, with respect to the SNP rs12435998 genotype.The SNP rs12435998 was prevalent in anaplastic and malignant gliomas, and in meningiomas of all histologic grades, but unrelated to brain tumor risks. In GBM patients, the SNP rs12435998 was associated with prolonged overall survival (OS) and better response to TMZ-based radio-chemotherapy. GBM stem cells with this SNP showed lower levels of SEL1L expression and enhanced sensitivity to VPA.     

未折叠蛋白反应与上皮间质转化在肿瘤中的研究进展

内质网是新合成的蛋白质进入分泌途径的细胞器。受多种生理及病理因素影响，细胞的分泌能力被扰乱，导致错误折叠和未折叠蛋白质的积累。进展期肿瘤细胞由于受到各种内外应激因子的刺激出现蛋白质稳态失衡和内质网应激。为了应对这种情况，肿瘤细胞通过激活未折叠蛋白反应维持生存。未折叠蛋白反应及上皮间质转化在肿瘤的发生、发展、转移中起到关键作用。这篇综述中，我们将分析未折叠蛋白反应在肿瘤中的作用，探讨内质网应激、未折叠蛋白反应与上皮间质转化之间的关系。     

Intracellular immunoglobulin in lymphocytes from patients with chronic lymphocytic leukemia: An immunoelectron microscopic study

The presence of ultrastructural distribution of intracellular immunoglobulin (Ig) in leukemic cells from patients with chronic lymphocytic leukemia (CLL) and in normal B cells were investigated by an immunoelectron microscopic technique. Most normal B cells had no intracellular Ig in spite of the presence of surface Ig. However, leukemic cells from 10 to 11 patients with CLL contained intracellular Ig and showed various staining patterns. In eight patients, Ig was present in the perinuclear space (PN), the endoplasmic reticulum (ER) and, if identified, the Golgi complexes. In four patients, most cells had diffuse staining of the cytoplasm. In five patients, Ig was detected in the ER-associated structures or the vesicles, in addition to the PN and ER. These findings suggest that CLL cells have a greater capacity to produce Ig than those of normal B cells and include various clones with distinct staining patterns of intracellular Ig.     

Tie2/TEK modulates the interaction of glioma and brain tumor stem cells with endothelial cells and promotes an invasive phenotype

Malignant gliomas are the prototype of highly infiltrative tumors and this characteristic is the main factor for the inevitable tumor recurrence and short survival after most aggressive therapies. The aberrant communication between glioma cells and tumor microenvironment represents one of the major factors regulating brain tumor dispersal. Our group has previously reported that the tyrosine kinase receptor Tie2/TEK is expressed in glioma cells and brain tumor stem cells and is associated with the malignant progression of these tumors. In this study, we sought to determine whether the angiopoietin 1 (Ang1)/Tie2 axis regulates crosstalk between glioma cells and endothelial cells. We found that Ang1 enhanced the adhesion of Tie2-expressing glioma and brain tumor stem cells to endothelial cells. Conversely, specific small interfering RNA (siRNA) knockdown of Tie2 expression inhibited the adhesion capability of glioma cells. Tie2 activation induced integrin β1 and N-cadherin upregulation, and neutralizing antibodies against these molecules inhibited the adhesion of Tie2-positive glioma cells to endothelial cells. In 2D and 3D cultures, we observed that Ang1/Tie2 axis activation was related to increased glioma cell invasion, which was inhibited by using Tie2 siRNA. Importantly, intracranial co-implantation of Tie2-positive glioma cells and endothelial cells in a mouse model resulted in diffusely invasive tumors with cell clusters surrounding glomeruloid vessels mimicking a tumoral niche distribution. Collectively, our results provide new information about the Tie2 signaling in glioma cells that regulates the cross-talk between glioma cells and tumor microenvironment, envisioning Tie2 as a multi-compartmental target for glioma therapy.