Trophic effect of various neuropeptides on the cultured ventral spinal cord of rat embryo

We studied the trophic effects of various neuropeptides, including substance P (SP), thyrotropin-releasing hormone (TRH), neurotensin (NT) and bombesin (BN) on explanted cultures of ventral spinal cord from 13- to 14-day-old rat embryos. In groups, receiving one type of drug only, there was a significant neurite-promoting effect (NPE) in SP, TRH and BN-treated cultures. At a concentration of 10(-12) M the most potent effect was shown by SP. However, BN had the most potent action at concentrations greater than 10(-10) M. It also became clear that there were maximum and minimum effective concentrations for these 3 neuropeptides. NT had no NPE at any concentration. There was no additional NPE when any two or all four of these neuropeptides were given simultaneously. Our results demonstrate that BN, SP, and TRH have a trophic effect on ventral spinal cord in cultures, and may contribute to a therapeutic strategy in amyotrophic lateral sclerosis.     

A porous 3D cell culture micro device for cell migration study

Cell migration under chemoattractant is an important biological step in cancer metastasis that causes the spread of malignant tumor cells. Porous polymeric materials are widely used to mimic the extracellular matrix (ECM) environment for applications such as three dimensional (3D) cell culturing and tissue engineering. In this paper we report a novel 3D cell culture device based on porous polymeric material to study cancer migration. We fabricated a porous channel on a polymeric chip using a selective ultrasonic foaming method. We demonstrate that a chemical concentration gradient could be established through the porous channel due to the slow diffusion process. We show that significant cell migration could be observed through the porous channel within 1–2 weeks of cell culturing when metastatic M4A4-GFP breast cancer cells were induced by 20% fetal bovine serum (FBS).We also developed a mathematical model to evaluate the diffusivity and concentration gradient through the fabricated porous structure.     

Autoantibodies to neurons and to the cytoskeleton in small cell carcinoma with paraneoplastic sensory neuropathy

Autoantibodies to neurons and to the cytoskeleton were demonstrated in the serum of a patient with small cell carcinoma of the lung (SCCL) and paraneoplastic sensory neuropathy. The serum reacted by immunofluorescence with the nuclei of neurons, and with cytochalasin B-sensitive "stress fibres" of cultured cells. The serum also reacted by immunofluorescence with the nuclei of some cultured cell lines. Immunoblotting experiments with brain tissue, with SCCL, HeLa and other cultured cells showed that the serum reacted with 1-4 bands of 35-39 kDa apparent m.w. Two dimensional immunoblotting showed that these molecules had a neutral pI. Antibodies, affinity-purified by elution from the 35-39 kDa bands, gave staining of the nuclei of neurons and of cultured cells and immunoblotted the same 35-39 kDa antigens. These observations show that the anti-neuronal autoantibody reacts not only with neurons and with SCCL but also with some cultured cell lines. Molecular mimicry has been invoked as the basis for the reactivity of this autoantibody with exposed epitopes on SCCL and on neurons.     

Chemotactic and mitogenic components of the alveolar macrophage response to particles ad neutrophil chemoattractant.

The pulmonary response to instilled particulates involves initial efflux of polymorphonuclear leukocytes (PMNs) and increased production of alveolar macrophages (AMs). The relationships of the cell migration and division components of the AM production to the initial PMN response after intrapulmonary carbon or latex administration are examined. Supernatants of lung lavages taken during early PMN migration to the alveoli promote sequential migration of PMNs, migration of monocytes, and division of pulmonary interstitial cells in normal mice. Supernatants taken during the phase of increased cell division in the lung produce no such effects, implying tht factors responsible for chemotaxis and mitosis are generated rapidly and are short-lived. A possible source is the interaction of AMs with particles, since supernatants of such incubations induce an inflammatory response in vivo. Similarly, when a synthetic chemoattractant is used, efflux of PMNs is followed by macrophages arising from migration of monocytes and from division of interstitial cells. The results suggest that particulate instillation in the lung stimulates a standard inflammatory response in which rapid generation of a factor(s) chemotactic for PMNs also attracts mononuclear cells to the alveoli. The initial efflux of cells may be explained by migration from the blood, but continued demand or replacement requires mitotic activity of precursors. For the alveolar macrophages, this includes division of cells in the pulmonary interstitium.     

Autoantibodies to Neurons and to the Cytoskeleton in Small Cell Carcinoma with Paraneoplastic Sensory Neuropathy

Autoantibodies to neurons and to the cytoskeleton were demonstrated in the serum of a patient with small cell carcinoma of the lung (SCCL) and paraneoplastic sensory neuropathy. The serum reacted by immunofluorescence with the nuclei of neurons, and with cytochalasin B-sensitive “stress fibres” of cultured cells. The serum also reacted by immunofluorescence with the nuclei of some cultured cell lines. Immunoblotting experiments with brain tissue, with SCCL, HeLa and other cultured cells showed that the serum reacted with 1–4 bands of 35–39 kDa apparent m.w. Two dimensional immunoblotting showed that these molecules had a neutral pI. Antibodies, affinity-purified by elution from the 35–39 kDa bands, gave staining of the nuclei of neurons and of cultured cells and immunoblotted the same 35–39 kDa antigens. These observations show that the anti-neuronal autoantibody reacts not only with neurons and with SCCL but also with some cultured cell lines. Molecular mimicry has been invoked as the basis for the reactivity of this autoantibody with exposed epitopes on SCCL and on neurons.     

Human chorionic gonadotropin in lung and lung tumors. Immunohistochemical study on unbalanced distribution of subunits

To demonstrate unbalanced distribution of subunits of human chorionic gonadotropin (hCG) in the lung and lung tumors and to clarify its significance in differentiation and carcinogenesis of the lung, immunohistochemistry was performed on human fetus, infant, and adult lungs, and endocrine and nonendocrine tumors of the lung. Tissues were immunostained for alpha-subunits and for beta-subunits of glycoprotein hormones (hCG, luteinizing hormone, follicle stimulating hormone, and thyroid stimulating hormone), serotonin, and gastrin-releasing peptide. Immunoreactive alpha-subunit was first identified in endocrine-like cells at the 39th gestational week, and was found in all infant lungs and two-thirds of adult lungs. The hCG beta-immunoreactive cells were extremely rare in an adult lung, and were not found in fetus or infant lungs. The alpha-subunit-containing cells were present in neuroepithelial bodies, tumorlets, carcinoid tumors, and small cell carcinomas of the lung (SCCL). There were occasionally alpha-subunit-containing cells in non-SCCL but one of the carcinomas also contained many serotonin-positive and gastrin-releasing peptide-positive cells in the same region. All alpha-subunit-immunoreactive cells lacked immunoreactivity for beta-subunits of glycoprotein hormones, except some for hCG beta in one carcinoid tumor. Immunoreactive cells for isolated hCG beta appeared much more frequently in non-SCCL than in SCCL. Most non-SCCL containing hCG beta-positive cells did not show alpha-subunit-immunoreactivity. Thus, immunohistochemical distribution of hCG-subunits was unbalanced and hCG-subunits may be expressed through an independent mechanism, commonly in the lung and lung tumors. The significance of isolated alpha-subunit is further discussed in light of multidirectional differentiation of lung neoplasms (14, 17).     

A comparative study of vascular proliferation in brain metastasis of lung carcinomas

Because of the marked vascular proliferation seen in brain metastases of small cell carcinoma of the lung (SCCL), we studied the morphometric and immunohistochemical characteristics of proliferating vessels in metastases from 20 autopsy cases of SCCL with brain metastasis. These were compared with those in surgically resected brain metastases of lung carcinomas, including 6 cases of SCCL, 19 cases of adenocarcinoma and 5 cases of squamous cell carcinoma. Angiogenesis in the tumours was scored by the microscopic angiogenesis grading system (MAGS). The MAGS score for autopsy and surgical metastatic lesions was highest in SCCL. Histologically, many vascular glomeruloid structures were formed in the brain metastases of SCCL, and immunohistochemistry revealed that these lesions were composed of proliferating endothelial cells and pericyte/smooth muscle cells. Immunostaining for basic fibroblast growth factor, a potent angiogenic factor, showed immunoreactivity in the tumour cells, regardless of histological type, and in the surrounding glial cells. Complex autocrine and paracrine phenomena participate in the development of metastatic cerebral lesions with vascular proliferation.     

Monoclonal antibody directed against neuroendocrine properties of both normal and malignant cells

A monoclonal antibody, 6H7, was produced by the immunization of small cell carcinoma of the lung (SCCL). Immunohistochemical examination indicated that 6H7 reacted not only with SCCL but also various neuronal and/or endocrine tumors such as neuroblastoma, pheochromocytoma, carcinoid and adrenal cortical tumors. 6H7 was also reactive with normal neuroendocrine tissues including brain, spinal cord, thyroid follicular cells, pancreatic islet cells and adrenal cells. 6H7 did not react with squamous cell carcinomas, one large cell carcinoma or most adenocarcinomas of the lung, or carcinomas of the stomach, colon, pancreas, breast and esophagus. The antigen recognized by 6H7 was analyzed on gel filtration after purification of the antigen by liquid chromatography which indicated the molecular weight of the antigen to be 270,000-300,000. From SDS-PAGE analysis the antigen reactive with 6H7 appeared to consist of polypeptide dimers of 128,000.     

Degree of neutrophil chemotaxis is dependent upon the chemoattractant and barrier.

Stimulated neutrophil migration across lung endothelial and epithelial barriers is important in lung inflammatory processes. To better understand the interaction between chemoattractants, neutrophils, and endothelium and epithelium, we compared the ability of leukotriene B4 (LTB4), formylmethionylleucylphenylalanine (FMLP), and platelet-activating factor (PAF) to induce human neutrophil migration across 3-microns-pore filters alone and human umbilical vein endothelial (HUVE) cells and two different epithelial cell types, Madin-Darby canine kidney (MDCK) cells and human lung A549 cells, cultured in monolayers on these filters. LTB4, FMLP, and PAF induced neutrophil migration through naked filters, endothelial cells, and epithelial cells in a dose-related fashion. At optimal chemoattractant doses, LTB4, FMLP, and PAF induced relatively equivalent neutrophil migration through filters and endothelial and epithelial monolayers. However, the doses at which optimal neutrophil migration was observed to occur as well as the degree of neutrophil migration through the three barriers varied depending upon the chemoattractant. Based on dose-response experiments, the relative rank order of potency for the three chemoattractants was: LTB4 = FMLP greater than PAF for filter alone barrier; LTB4 greater than FMLP greater than PAF for HUVE cell barrier; and FMLP greater than LTB4 greater than PAF for MDCK and A549 epithelial cell barriers. Our data suggest that neutrophil chemotactic and subsequent lung inflammatory responses are interrelatedly influenced by both the quantity and type of chemoattractant present and the barrier through which the neutrophil must migrate.     

Inhibition of tumor invasion and extracellular matrix degradation by ubenimex (bestatin)

We have investigated the effect of the immunomodulator ubenimex (hereafter referred to as bestatin) on the enzymatic degradation of the extracellular matrix by human renal cell carcinoma SN12M cells during the invasive process. The invasion of SN12M cells into reconstituted basement membrane (Matrigel) was inhibited by the presence of bestatin in a concentration-dependent manner. However, bestatin did not have any effect on tumor cell adhesion and migration to the extracellular matrices which may be involved in tumor cell invasion. Bestatin inhibited the degradation of type IV collagen by tumor cells, but not by tumor-conditioned medium (TCM), in a concentration-dependent manner. We also found that bestatin inhibited hydrolysing activities towards substrates of aminopeptidases in SN12M cells. Since bestatin was found to inhibit aminopeptidase activity, the inhibition of tumor invasion by bestatin is likely to be associated with its action as an enzyme inhibitor. Bestatin only slightly inhibited tumor cell plasmin activity, which can lead to the conversion of the latent collagenase to the active form, but this slight effect was not significant. The zymography of TCM from SN12M cells showed that the treatment of tumor cells with bestatin resulted in the disappearance of the 68 kDa type IV collagenase-enzyme level (active form) and slight reduction of the 72 kDa type IV collagenase-enzyme level (latent form). These results indicated that bestatin may inhibit tumor cell invasion through a mechanism involving its inhibitory action on aminopeptidases in tumor cells, suggesting that the aminopeptidase may partly be associated with the conversion of a latent form of type IV procollagenase to an active form or the secretion of the collagenases from tumor cells.     

Low density lipoproteins and Lovastatin modulate the organ-specific transendothelial migration of primary and metastatic human colon adenocarcinoma cell lines in vitro

Tumor cell arrest and tumor migration are two of the critical steps in the metastatic cascade. We hypothesized that these steps may be facilitated by the low density lipoprotein (LDL)-induced activation of microvessel endothelial cells (MVEC). The purpose of our study was to investigate the biological effects of an LDL-enriched milieu and the effects of the anticholesterol drug Lovastatin on metastatic behavior. The SW480 and SW620 are primary and metastatic human colonic adenocarcinoma cell lines derived from the same patient. We investigated the effect of LDL on adhesion and migration of the two tumor cell lines across human brain, lung, liver and dermal endothelial monolayers. Adhesion and migration assays were done before and after pretreat-ment of the MVEC or tumor cells with LDL (100 mg/ml) for 24 h. Although metastatic SW620 cells were more adherent to MVEC compared with primary SW480 cells, LDL pretreatment of SW480 and SW620 cells did not affect tumor cell adhesion to MVEC. In contrast, tumor cell migration was significantly increased across endothelial monolayers when MVEC were pretreated with LDL. Transendothelial cell migration was not sig-nificantly affected by pretreatment of the tumor cells with LDL. Lovastatin is an inhibitor of HMG-CoA reduc-tase, the rate-limiting enzyme in cholesterol biosynthesis. It has been shown to have anti-tumor activity in vitro. We investigated the effect of Lovastatin on tumor cell kinetics and tumor cell migration across MVEC. Growth curves and migration assays were done before and after pretreatment of the tumor cells with Lovastatin (30 mg/ml). Migration assays were also done after treatment of unstimulated or LDL-stimulated MVEC (100 mg/ml) for 24 h with Lovastatin. Lovastatin inhibited the in vitro growth of the metastatic SW620 cell line to a greater extent than the invasive SW480E cell line. On the other hand, pretreatment of tumor cells with Lovastatin (30 mg/ml) did not suppress transendothelial tumor cell migration of tumor cells. Finally, Lovastatin given to mice effectively suppressed the number of MCA-26 tumor colonies in the liver of Balb/c mice com-pared with untreated mice. ©Lippincott Williams & Wilkins     

Neuroendocrine Carcinoma of Skin with Simultaneous Cytokeratin Expression

An unusual tumor of the skin was removed from the thigh of a 52-year-old white male. By light microscopy, the tumor was composed of intermediate and small cells in sheets and clusters. Ultrastructural study of the tumor cells showed numerous dense core granules and dendritic cell processes as well as intermediate filaments and cell junctions frequently within the same cells. Most of the tumor cells were stained intensely by antibodies to neurone-specific enolase (NSE), a marker of cells of the central and peripheral nervous system. The neuropeptides met-enkephalin and vasoactive intestinal peptide (VIP) were also found in tumor cells. Immunohistochemistry furthermore demonstrated cytokeratin. Both the ultrastructural appearance and keratin content of this tumor set it apart from conventional Merkel cell (or trabecular) carcinoma of the skin in a manner analogous to bipartite (i.e., epidermoid and small cell) carcinoma of lung. The production of neuropeptides simultaneously with the production of keratin establishes this as a bipartite skin tumor (i.e., ectodermal and neuroectodermal phenotype). We suggest that at least some primary neuroendocrine tumors of the skin arise from multi-potential ectodermal cells not of neural crest origin, as has been proposed for small cell carcinoma of lung.     

Soluble factors released by the target organ enhance the urokinase-type plasminogen activator activity of metastatic tumor cells

The ability of tumor cells to respond to microenvironmental factors present in the target organ may be necessary for successful metastasis. Many studies suggest that urokinase-type plasminogen activator (u-PA) has a significant role in several steps of the metastatic process. In previous work it had been observed that lung conditioned media stimulated the migration and growthin vitro of cells from a murine mammary adenocarcinoma (M3) with moderate lung metastasizing potential. In the same experiments liver conditioned medium exerted a marked cytostatic effect on M3 cells. The aim of the present work to investigate whether conditioned media from lung, kidney or liver, were able to modulate u-PAin vitro secretion by these murine M3 cells. Secreted u-PA measured by fibrinolytic assay, was significantly increased only when M3 primary cultured cells were treated for 24 h with lung conditioned media prepared from normal mice or from mice bearing a small tumor. Exposure to kidney or liver conditioned media did not modify the u-PA secretion pattern already shown by the tumor cells. The activity shown by lung conditioned media seemed to be specific for these syngeneic tumor cells, as no effect was observed on murine embryo cells. These results suggest that soluble factors released by the target organ could specifically induce tumor cellsin vivo to enhance the production of degradative enzymes, thus facilitating the last steps of the metastatic cascade.     

Neuroendocrine carcinoma of skin with simultaneous cytokeratin expression

An unusual tumor of the skin was removed from the thigh of a 52-year-old white male. By light microscopy, the tumor was composed of intermediate and small cells in sheets and clusters. Ultrastructural study of the tumor cells showed numerous dense core granules and dendritic cell processes as well as intermediate filaments and cell junctions frequently within the same cells. Most of the tumor cells were stained intensely by antibodies to neurone-specific enolase (NSE), a marker of cells of the central and peripheral nervous system. The neuropeptides met-enkephalin and vasoactive intestinal peptide (VIP) were also found in tumor cells. Immunohistochemistry furthermore demonstrated cytokeratin. Both the ultrastructural appearance and keratin content of this tumor set it apart from conventional Merkel cell (or trabecular) carcinoma of the skin in a manner analogous to bipartite (i.e., epidermoid and small cell) carcinoma of lung. The production of neuropeptides simultaneously with the production of keratin establishes this as a bipartite skin tumor (i.e., ectodermal and neuroectodermal phenotype). We suggest that at least some primary neuroendocrine tumors of the skin arise from multipotential ectodermal cells not of neural crest origin, as has been proposed for small cell carcinoma of lung.     

Neuroendocrine Carcinoma of Skin with Simultaneous Cytokeratin Expression

An unusual tumor of the skin was removed from the thigh of a 52-year-old white male. By light microscopy, the tumor was composed of intermediate and small cells in sheets and clusters. Ultrastructural study of the tumor cells showed numerous dense core granules and dendritic cell processes as well as intermediate filaments and cell junctions frequently within the same cells. Most of the tumor cells were stained intensely by antibodies to neurone-specific enolase (NSE), a marker of cells of the central and peripheral nervous system. The neuropeptides met-enkephalin and vasoactive intestinal peptide (VIP) were also found in tumor cells. Immunohistochemistry furthermore demonstrated cytokeratin. Both the ultrastructural appearance and keratin content of this tumor set it apart from conventional Merkel cell (or trabecular) carcinoma of the skin in a manner analogous to bipartite (i.e., epidermoid and small cell) carcinoma of lung. The production of neuropeptides simultaneously with the production of keratin establishes this as a bipartite skin tumor (i.e., ectodermal and neuroectodermal phenotype). We suggest that at least some primary neuroendocrine tumors of the skin arise from multi-potential ectodermal cells not of neural crest origin, as has been proposed for small cell carcinoma of lung.     

Mesothelial cells produce a chemoattractant for lung fibroblasts: role of fibronectin

Pleural fibrosis may complicate several types of non-exudative pleural injury. Although the pathogenesis of such lesions is poorly understood, it is conceivable that mesothelial cells may recruit fibroblasts to sites of pleural damage. In order to test this possibility, conditioned medium from cultured rat mesothelial cells was tested for chemoattractant activity towards RL-87 rat lung fibroblasts. For this purpose, rat pleural or pericardial mesothelial cells were maintained in vitro for 6 to 96 h. Conditioned medium from each source was obtained at defined culture times and tested for chemotactic activity in a 48-well microchemotaxis assembly. A progressive, time-dependent increase in fibroblast chemoattractant activity was detected in both pleural and pericardial mesothelial cell conditioned medium samples. This effect was maximal in 96-h cultures. Checkerboard analysis revealed that the conditioned medium was truly chemotactic for lung fibroblasts. Characterization of the chemoattractant demonstrated that it was a nondialyzable (greater than 16 kD), thermolabile (100 degrees C for 15 min), acid-stable (pH 2.5), trypsin-sensitive, and pepsin-sensitive protein. The chemotaxin was shown to be fibronectin, since activity was abolished, in a dose-dependent manner, by treatment with anti-rat fibronectin antiserum as well as by passage through a gelatin agarose affinity column. This product consisted of two bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis of apparent molecular masses 250 and 220 kD. The secretion of a mesothelial cell-derived fibroblast chemoattractant may play a role in the response of the pleura to injury and in the pathogenesis of pleural fibrosis.     

IL-4 and IL-13 Induce Chemotaxis of Human Foreskin Fibroblasts, But Not Human Fetal Lung Fibroblasts

Through shared receptors, IL-4 and IL-13 have been suggested to regulate not only inflammatory cells, but also to play a role in stimulating fibroblasts during fibrotic processes. Previous studies have shown that IL-4 is a chemoattractant for foreskin fibroblasts. The current study was designed to determine the effect of IL-4 and IL-13 on the migration of two types of fibroblasts: foreskin and human fetal lung fibroblasts (HFL-1). Using the Boyden blindwell chamber method, human foreskin or fetal lung fibroblasts (both 10(6)/mL) were placed in upper wells with various concentrations of IL-4 or IL-13 in the lower wells as chemoattractants. Both IL-4 (1 pg/mL) and IL-13 (100 pg/mL) induced foreskin fibroblast chemotaxis, up to 50 +/- 8 and 24 +/- 7 cells/5 high-power fields, respectively (both p < 0.05). In contrast, neither cytokine induced migration of the lung fibroblasts although both type of cells express IL-4 receptor and IL-13alpha1 receptor. These results suggest that fibroblasts are heterogeneous with regard to their ability to respond to cytokine-driven chemotaxis. Therefore, the role of specific cytokines in mediating fibrotic responses might vary depending on local mesenchymal cell responses.     

长链非编码MALAT-1 调控miR-205 的表达影响非小细胞肺癌细胞的生长和迁移

目的:探讨在非小细胞肺癌中,LncRNA MALAT-1 与miR-205 的相互关系,以及影响肺癌细胞生物学行为的机制。方法:qPCR 检测不同非小细胞肺癌中LncRNA MALAT-1 的表达情况;双荧光素酶报告基因检测MALAT-1 与miR-205的相互作用;Transwell 侵袭实验和划痕实验检测抑制MALAT-1 后肺癌细胞侵袭能力的变化,以及抑制miR-205 的表达后肺癌细胞迁移和侵袭能力的恢复情况;裸鼠皮下成瘤检测抑制LncRNA MALAT-1 后肺癌细胞体外成瘤体积和质量变化。结果:与其他肺癌细胞株相比,A549 细胞中MALAT-1 表达最高,miR-205 的表达水平最低;双荧光素酶实验证实MALAT-1 能与miR-205 的3忆UTR 特异性结合,可以调控miR-205 的表达与活性;抑制MALAT-1 的表达后可以降低肺癌细胞的迁移和侵袭能力;抑制miR-205 的表达水平过后,肺癌细胞的迁移和侵袭能力相对增强;抑制MALAT-1 的表达后,荷瘤小鼠的肿瘤体积和重量都明显减小。结论:MALAT-1 可以调控miR-205 的表达影响肺癌细胞A549 的侵袭和迁移能力。     

Apelin triggers macrophage polarization to M2 type in head and neck cancer

Cancer comes after cardiovascular diseases in terms of mortality rate in the world. Chemotherapy, radiotherapy and surgical interventions are the current cancer treatment. Recently, it has been observed that immunotherapeutic approaches provide a significant improvement when used along with these interventions. The mononuclear system mainly consists of macrophages that play an active role in the pathology of many diseases because of having high plasticity capacities. Previous research suggested that they can be used as an alternative to cancer treatment. Aim was to investigate the effect of apelin on macrophage polarization in the tumor microenvironment.Mouse macrophage cell line RAW264.7 cells and head and were chosen for this study. The apelin expression was knockdown in neck cell carcinoma cell line SCCL MT1 cells using shRNA technique. SCCL MT1 cells having normal or suppressed apelin expression were co-cultured with mouse macrophage RAW264.7 cells. The effect of co-culturing on the expression of inflammatory genes in RAW264.7 cells was investigated.Suppressed apelin expression in SCCL MT1 cells resulted in elevated pro-inflammatory response in co-cultured macrophages. Expression of the IL1β, IL6, and TNFα genes significantly increased, however anti-inflammatory cytokine levels were significantly decreased. However, in the control group, a downregulation was determined in pro-inflammatory genes, while an increase was observed in anti-inflammatory genes. The protein levels of these cytokines in concordance with the RT-PCR analysis.As a result of this study, apelin released from cancer cells was found to affect macrophage polarization. These results indicated that the apelin peptide may cause the intense presence of M2-type macrophages in the tumor niche, and the therapeutic approaches targeting of apelin in cancer cells may have a potential role in macrophage polarization.