Induction of TNF-alpha,uPA, IL-8 and MCP-1 by doxorubicin in human lung carcinoma cells

Purpose We have previously demonstrated doxorubicin-induced urokinase (uPA) and interleukin-8 (IL-8) expression in human H69 small-cell lung carcinoma (SCLC) cells by a microarray technique using Human Cancer Chip version 2, in which 425 human "cancer-related" genes are spotted on the plates. The microarray analysis also revealed a significant induction of tumor necrosis factor-alpha (TNF-), and doxorubicin-induced macrophage chemoattractant protein-1 (MCP-1) expression was demonstrated by an RNase protection assay. We extended the study by testing the effects of doxorubicin on the induction of TNF-, uPA, IL-8 and MCP-1 in other types of lung carcinoma cells.Methods We investigated the effects of doxorubicin on the expression of TNF-, uPA, IL-8 and MCP-1 in 12 human lung carcinoma cell lines, including five SCLC, three adenocarcinoma and four squamous cell carcinoma cells. The surface expression of their receptors was also investigated.Results TNF- was significantly induced in three cell lines, H69, SBC-7 (SCLC) and PC-9 (adenocarcinoma), uPA in five cell lines, H69, SBC-7, EBC-1 (squamous cell), EBC-2 (squamous cell), and Sq-1 (squamous cell), IL-8 in three cell lines, H69, PC-9 and EBC-1, and MCP-1 in five cell lines, H69, SBC-3 (SCLC), SBC-7, PC-9 and Sq-1. In H69 cells, TNF- antigen levels were increased approximately fivefold in the conditioned medium of doxorubicin-treated cells, in parallel with an increase in mRNA levels. As with uPA and IL-8, the maximum induction was observed at the "sublethal" concentrations of 2 and 4 M at which cell growth was slightly inhibited 24 h after treatment. Furthermore, the cells did not express receptors including types I and II TNF- receptors, uPA receptor (uPAR), C-x-C-chemokine receptor-1 (CXCR-1), or C-C-chemokine receptor-2, corresponding to TNF-, uPA, IL-8 and MCP-1, respectively, that were induced by doxorubicin in the cells, although SBC-7 cells expressed uPAR, and EBC-1 cells expressed CXCR-1.Conclusions TNF-, uPA, IL-8 and MCP-1 induced and secreted from tumor cells upon doxorubicin stimulation may activate surrounding cells expressing the receptors such as neutrophils and monocytes/macrophages in a paracrine fashion. TNF- is a major proinflammatory cytokine, and IL-8 and MCP-1 are major chemoattractants for neutrophils and monocytes/macrophages, respectively. Furthermore, uPA activates matrix metalloproteinase 9 which can truncate and activate IL-8. Thus, the simultaneous induction of TNF-, uPA, IL-8 and MCP-1 may enhance the interaction between tumor and inflammatory/immune cells, and augment cytotoxicity.Abbreviations BSA Bovine serum albumin - C-C Cysteine-cysteine - C-x-C Cysteine-x-cysteine - CCR-2 C-C chemokine receptor-2 - CXCR-1 C-x-C chemokine receptor-1 - ELISA Enzyme-linked immunosorbent assay - IL-1 Interleukin-1 - IL-6 Interleukin-6 - IL-8 Interleukin-8 - MCP-1 Macrophage chemoattractant protein-1 - MTT (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide - PBS Phosphate-buffered saline - ROS Reactive oxygen species - SCLC Small-cell lung cancer - TNF- Tumor necrosis factor-alpha - TNFRI Type I tumor necrosis factor-alpha receptor - TNFRII Type II tumor necrosis factor-alpha receptor - uPA Urokinase-type plasminogen activator - uPAR Urokinase-type plasminogen activator receptor     

The expression of tissue factor correlates with proliferative ability in meningioma

Tissue factor (TF) is a cell-surface glycoprotein responsible for initiating the extrinsic pathway of coagulation; this is inhibited by tissue factor pathway inhibitor (TFPI). As TF reportedly regulates tumor growth and angiogenesis, we investigated the role of TF in meningioma. Using immunohistochemical methods, we studied the expression of TF, TFPI, MIB-1 labeling index in 44 meningiomas to determine whether those factors reflect histological grade and proliferative ability. CD31 and CD68 immunostaining was used to assess vascular density and macrophage infiltration, respectively. Additionally we assessed the influence of TF on meningioma cell proliferation by MTT assay. TF was expressed in 1 of 34 (2.9%) benign, 1 of 5 (20%) atypical, and all of 5 anaplastic meningiomas. TFPI was detected in 2 benign (5.9%), 1 atypical (20%), and 3 (60%) anaplastic meningiomas. Both TF and TFPI expression was significantly correlated with the MIB-1 labeling index (LI). However, neither TF nor TFPI showed a correlation with vascular density. The density of tumor-associated macrophages was not correlated with TF or TFPI immunoreactivity. MTT assay revealed that TF increased the proliferation of meningioma cells. Although some macrophages expressed TF, a great number of the TF immunopositive parenchymal cells in the meningiomas were tumor cells. The present study suggest that the TF system reflects the proliferative ability and malignancy of meningiomas.     

Expression of serine proteinase inhibitor PP5/TFPI-2/MSPI decreases the invasive potential of human choriocarcinoma cells in vitro and in vivo

OBJECTIVE: PP5/TFPI-2/MSPI is a Kunitz-type serine proteinase inhibitor with broad inhibitory spectra, abundantly produced by placenta and detected in the blood of pregnant women. Expression of PP5/TFPI-2/MSPI is exclusively detected in syncytiotrophoblasts of placenta, but is barely detectable in choriocarcinoma cells, a trophoblast-derived malignant tumor. Chromosome 7, in which the PP5/TFPI-2/MSPI gene is localized, is frequently lost in various types of tumors. We attempted to elucidate the relation between PP5/TFPI-2/MSPI expression and the malignant properties of choriocarcinoma cells. METHODS: Human choriocarcinoma cells, JAR, were transfected with either a human PP5/TFPI-2/MSPI expression vector or an empty vector, and stable clones were obtained. Messenger RNA expression, protein secretion/localization, growth rate, and plating efficiency were evaluated. In vitro migration and invasive activity were determined by transwell chamber experiments. In vivo tumor growth was evaluated by the subcutaneous injection of cells to nude mice and followed by histological examination. RESULTS: Expression of mRNA and protein of PP5/TFPI-2/MSPI were confirmed, and a high producing clone and a low producing clone were chosen for further analysis. The majority of secreted PP5/TFPI-2/MSPI protein was revealed to associate with the extracellular matrix. Expression of PP5/TFPI-2/MSPI did not affect the growth and migration of the tumor cells, but enhanced their plating efficiency. Its expression significantly inhibited invasion through the Matrigel. Invasive growth into the subcutaneous muscle layer was not evident in the nude mouse tumors of the PP5/TFPI-2/MSPI-expressing cells. CONCLUSION: PP5/TFPI-2/MSPI-expressing choriocarcinoma cells showed suppressed potential of invasion in vitro and in vivo. It is suggested that loss or suppression of PP5/TFPI-2/MSPI expression may result in the acquisition of invasiveness in choriocarcinoma cells.     

Hypothalamic neuroactivity in specific processes and central regulation of body temperature and water intake

The method described was designed to elucidate the role of a particular neuronal system or specific nucleus in the central nervous system (CNS) in controlling physiological and biological functions. The neurochemical aspects of the CNS regulatory mechanism and related networks remain to be further investigated. There is little information available about the relationship between neuroactivity in the specific brain nuclei and physiological or biological responses in mammals. An adequate analysis of this relationship provides valuable insight to clarify which nucleus and what types of neurons are truly involved in the excitation of physiological events and its regulation. In the present study, we used microdialysis for stimulation of the anterior hypothalamus (AH) and simultaneous analysis of cholinergic activity, and we investigated c-Fos-like immunoreactivity (Fos-IR) in the brain in the same animal following microdialysis. The nuclear protein c-Fos, the product of c-fos oncogene, has been used as a marker of neuronal activity at the cellular level in the brain. Various physiological and pharmacological stimuli have been shown to induce Fos-IR in specific neuronal populations located in various regions of the brain. However, there are few studies investigating the responses produced by c-Fos expression in specific regions in same animals. We showed the involvement of hypothalamic cholinergic mechanisms in the thermoregulatory and water regulatory processes using the above procedures.     

A case of adenoid cystic carcinoma (acc) of the Breast and review of the utility of preoperative imaging diagnose

A case of adenoid cystic carcinoma (ACC) of the breast in a 66-year-old woman is reported herein. ACC accounts for about 0.1% of all breast cancers. Our patient presented with a small, elastic and hard mass, measuring 2.0x2.0 cm, between both outer quadrants of the right breast. Although physical examination, ultrasonography and magnetic resonance (MR) mammography suggested a benign tumor, aspiration biopsy cytology (ABC) was performed twice, and the second ABC specimen was evaluated as suspicious for breast carcinoma. Breast conserving surgery with a level II lymph node dissection was subsequently performed. There was no lymph node metastases and estrogen receptor (ER) status was negative. Light microscopy revealed various growth patterns, with the cells showing biphasic cellularity. According to immunohistochemical analyses, CEA, actin and vimentin were positive, S-100 protein was negative, and the cytokeratin reaction was partially positive. Therefore, ACC of the breast was diagnosed. Although ACC of the breast is a rare neoplasm, it should be considered in the differential diagnosis even if various diagnostic imaging studies suggest a benign tumor of the breast. Awareness of this tumor will help prevent misdiagnosis.     

Single-incision laparoscopic surgery for benign multicystic mesothelioma of the peritoneum in a young man: A case report

Benign multicystic peritoneal mesothelioma (BMPM) is a rare condition, particularly in men, and the preoperative diagnosis poses a challenge. Here, we present a case involving single-incision laparoscopic surgery (SILS) for BMPM in a 24-year-old man with a pelvic mass and a history of ulcerative colitis. Pelvic imaging revealed multifocal cysts, prompting the performance of SILS. The tumor was successfully resected with no residual lesions, and pathology confirmed the diagnosis of BMPM. This case represents the first documented instance of SILS being employed for BMPM in a man. BMPM, characterized by pelvic multifocal cysts, is a differential diagnosis, and SILS emerges as a viable option for both diagnosis and treatment.     

Speciation of arsenic in biological samples

Speciation of arsenicals in biological samples is an essential tool to gain insight into its distribution in tissues and its species-specific toxicity to target organs. Biological samples (urine, hair, fingernail) examined in the present study were collected from 41 people of West Bengal, India, who were drinking arsenic (As)-contaminated water, whereas 25 blood and urine samples were collected from a population who stopped drinking As contaminated water 2 years before the blood collection. Speciation of arsenicals in urine, water-methanol extract of freeze-dried red blood cells (RBCs), trichloroacetic acid treated plasma, and water extract of hair and fingernail was carried out by high-performance liquid chromatography (HPLC)-inductively coupled argon plasma mass spectrometry (ICP MS). Urine contained arsenobetaine (AsB, 1.0%), arsenite (iAs(III), 11.3), arsenate (iAs(V), 10.1), monomethylarsonous acid (MMA(III), 6.6), monomethylarsonic acid (MMA(V), 10.5), dimethylarsinous acid (DMA(III), 13.0), and dimethylarsinic acid (DMA(V), 47.5); fingernail contained iAs(III) (62.4%), iAs(V) (20.2), MMA(V) (5.7), DMA(III) (8.9), and DMA(V) (2.8); hair contained iAs(III) (58.9%), iAs(V) (34.8), MMA(V) (2.9), and DMA(V) (3.4); RBCs contained AsB (22.5%) and DMA(V) (77.5); and blood plasma contained AsB (16.7%), iAs(III) (21.1), MMA(V) (27.1), and DMA(V) (35.1). MMA(III), DMA(III), and iAs(V) were not found in any plasma and RBCs samples, but urine contained all of them. Arsenic in urine, fingernails, and hair are positively correlated with water As, suggesting that any of these measurements could be considered as a biomarker to As exposure. Status of urine and exogenous contamination of hair urgently need speciation of As in these samples, but speciation of As in nail is related to its total As (tAs) concentration. Therefore, total As concentrations of nails could be considered as biomarker to As exposure in the endemic areas.