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Purpose@#mTORC1 and mTORC2 inhibition by Ku-0063794 could confer profound anticancer effects against cancer cells because it eliminates feedback activation of Akt. Herein, we aimed to determine anticancer effects of docetaxel and Ku-0063794, individually or in combination, against breast cancer cells, especially triple-negative breast cancer (TNBC) cells. @*Materials and Methods@#MCF-7 breast cancer and MDA-MB-231 TNBC cell lines for in vitro studies and mouse xenograft model for in vivo studies were used to investigate the effect of docetaxel, Ku-0063794, or their combination. @*Results@#In the in vitro experiments, combination therapy synergistically reduced cell viability and induced higher apoptotic cell death in breast cancer cells than the individual monotherapies (p < 0.05). Western blot analysis and flow cytometric analysis showed that the combination therapy induced higher apoptotic cell death than the individual monotherapies (p < 0.05). In the in vivo experiment, docetaxel and Ku-0063794 combination therapy reduced the growth of MDA-MB-231 cells xenografted in the nude mice better than in the individual monotherapies (p < 0.05). Immunohistochemistry showed that the combination therapy induced the highest expression of cleaved caspase-3 and the lowest expression of Bcl-xL in the MDA-MB-231 cells xenografted in the nude mice (p < 0.05). Western blot analysis and immunofluorescence, incorporating both in vitro and in vivo experiments, consistently validated that unlike individual monotherapies, docetaxel and Ku-0063794 combination therapy significantly inhibited epithelial-mesenchymal transition (EMT) and autophagy (p < 0.05). @*Conclusion@#These data suggest that docetaxel and Ku-0063794 combination therapy has higher anticancer activities over individual monotherapies against MDA-MB-231 TNBC cells through a greater inhibition of autophagy and EMT.
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Purpose@#Survivin is a typical antiapoptotic protein. It is copiously expressed during human fetal development but is infrequently present in adult tissues. In this experiment, we researched the treatment effect of the secretome that adiposederived stem cells (ASCs) transfected with survivin. @*Methods@#First of all, we generated survivin-overexpressing ASCs transfected with a plasmid comprising a gene encoding survivin. The secreted substances released from survivin-overexpressing ASCs (survivin-secretome) were collected, and were determined their in vitro and in vivo therapeutic potential, especially in the model of liver impairment. @*Results@#In vitro, the survivin-secretome significantly increased cell viability and promoted the expression of proliferationrelated markers (proliferating cell nuclear antigen [PCNA], phospho-signal transducer and activator of transcription 3 (p-STAT3), hepatocyte growth factor [HGF], vascular endothelial growth factor [VEGF]) and anti-apoptosis-related markers (myeloid cell leukemia-1 [Mcl-1] and survivin) (P < 0.05). In vivo using 70% hepatectomy mice, the survivin-secretome group exhibited the lowest serum levels of interleukin-6, tumor necrosis factor-α (P < 0.05). The serum levels of liver transaminases (alanine aminotransferase and aspartate aminotransferase) were also the lowest in the survivin-secretome group (P < 0.05). The survivin-secretome group also exhibited the highest liver regeneration on the 7th day after 70% partial hepatectomy (P < 0.05). In the subsequent liver specimen analysis, the specimens of survivin-secretome exhibited the highest expression of p-STAT3, HGF, VEGF, PCNA, and Mcl-1 and the lowest expression of bcl-2-like protein 4 (P < 0.05). @*Conclusion@#Taken together, secretome secreted by survivin-overexpressing ASCs could be an effective way to improve liver regeneration and repair for liver injury treatment.
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Background@#Di-2-ethylhexyl phthalate (DEHP) is known to disrupt thyroid hormonal status. However, the underlying molecular mechanism of this disruption is unclear. Therefore, we investigated the direct effects of DEHP on the thyroid gland. @*Methods@#DEHP (vehicle, 50 mg/kg, and 500 mg/kg) was administered to Sprague-Dawley rats for 2 weeks. The expression of the thyroid hormone synthesis pathway in rat thyroid tissues was analyzed through RNA sequencing analysis, quantitative reverse transcription-polymerase chain reaction (RT-PCR), and immunohistochemical (IHC) staining. DEHP was treated to FRTL-5 rat thyroid cells, and an RT-PCR analysis was performed. A reporter gene assay containing the promoter of thyroid stimulating hormone receptor (TSHR) in Nthy-ori 3-1 human thyroid cells was constructed, and luciferase activity was determined. @*Results@#After DEHP treatment, the free thyroxine (T4) and total T4 levels in rats significantly decreased. RNA sequencing analysis of rat thyroid tissues showed little difference between vehicle and DEHP groups. In the RT-PCR analysis, Tshr expression was significantly lower in both DEHP groups (50 and 500 mg/kg) compared to that in the vehicle group, and IHC staining showed that TSHR expression in the 50 mg/kg DEHP group significantly decreased. DEHP treatment to FRTL-5 cells significantly down-regulated Tshr expression. DEHP treatment also reduced luciferase activity in a reporter gene assay for TSHR. @*Conclusion@#Although overall genetic changes in the thyroid hormone synthesis pathway are not clear, DEHP exposure could significantly down-regulate Tshr expression in thyroid glands. Down-regulation of Tshr gene appears to be one of potential mechanisms of thyroid disruption by DEHP exposure.
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Background@#Di-2-ethylhexyl phthalate (DEHP) is known to disrupt thyroid hormonal status. However, the underlying molecular mechanism of this disruption is unclear. Therefore, we investigated the direct effects of DEHP on the thyroid gland. @*Methods@#DEHP (vehicle, 50 mg/kg, and 500 mg/kg) was administered to Sprague-Dawley rats for 2 weeks. The expression of the thyroid hormone synthesis pathway in rat thyroid tissues was analyzed through RNA sequencing analysis, quantitative reverse transcription-polymerase chain reaction (RT-PCR), and immunohistochemical (IHC) staining. DEHP was treated to FRTL-5 rat thyroid cells, and an RT-PCR analysis was performed. A reporter gene assay containing the promoter of thyroid stimulating hormone receptor (TSHR) in Nthy-ori 3-1 human thyroid cells was constructed, and luciferase activity was determined. @*Results@#After DEHP treatment, the free thyroxine (T4) and total T4 levels in rats significantly decreased. RNA sequencing analysis of rat thyroid tissues showed little difference between vehicle and DEHP groups. In the RT-PCR analysis, Tshr expression was significantly lower in both DEHP groups (50 and 500 mg/kg) compared to that in the vehicle group, and IHC staining showed that TSHR expression in the 50 mg/kg DEHP group significantly decreased. DEHP treatment to FRTL-5 cells significantly down-regulated Tshr expression. DEHP treatment also reduced luciferase activity in a reporter gene assay for TSHR. @*Conclusion@#Although overall genetic changes in the thyroid hormone synthesis pathway are not clear, DEHP exposure could significantly down-regulate Tshr expression in thyroid glands. Down-regulation of Tshr gene appears to be one of potential mechanisms of thyroid disruption by DEHP exposure.
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Purpose@#Visfatin is a key cytokine released from the pe ripheral blood mononuclear cells (PBMCs) as well as adipose tissue, and it is involved in immune response as well as inflammation. In this study, we investigated whether the serum visfatin level could be a prognostic factor for predicting the severity of inflammation in patients with acute cholecystitis. @*Methods@#We examined the blood samples and gallbladder specimens from patients who underwent laparoscopic cholecystectomy for either acute (n = 18) or chronic cholecystitis (n = 18). We determined the visfatin levels of these samples using various procedures such as real-time polymerase chain reaction, enzyme-linked immunosorbent assay, western blotting, and immunohistochemistry. @*Results@#The patients with acute cholecystitis exhibited higher mRNA expression of visfatin in PBMCs, higher serum levels of visfatin, and increased protein expression of visfatin in the gallbladder specimens than in patients with chronic cholecystitis. In the in vitro model of acute cholecystitis, the mRNA expression of visfatin showed the fastest increase among the other pro-inflammatory mediators studied, including interleukin (IL)-10, tumor necrosis factor-, IL-6, intracellular adhesion molecule-1, and ascular cell adhesion molecule-1. Inhibition of visfatin using siRNA abrogated the inhibitory effects of lipopolysaccharide (LPS) on the expression of ABCG1 in GBECs, suggesting that visfatin is significantly involved in the LPS-driven suppression of ABCG1. @*Conclusion@#Taken together, we concluded that visfatin is a pro-inflammatory mediators that is upregulated during acute cholecystitis and is expected to be increased within a short time after inflammation. Therefore, measuring the serum level of visfatin would be helpful in predicting the inflammatory severity in the patients with acute cholecystitis.
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Purpose@#Visfatin is a key cytokine released from the pe ripheral blood mononuclear cells (PBMCs) as well as adipose tissue, and it is involved in immune response as well as inflammation. In this study, we investigated whether the serum visfatin level could be a prognostic factor for predicting the severity of inflammation in patients with acute cholecystitis. @*Methods@#We examined the blood samples and gallbladder specimens from patients who underwent laparoscopic cholecystectomy for either acute (n = 18) or chronic cholecystitis (n = 18). We determined the visfatin levels of these samples using various procedures such as real-time polymerase chain reaction, enzyme-linked immunosorbent assay, western blotting, and immunohistochemistry. @*Results@#The patients with acute cholecystitis exhibited higher mRNA expression of visfatin in PBMCs, higher serum levels of visfatin, and increased protein expression of visfatin in the gallbladder specimens than in patients with chronic cholecystitis. In the in vitro model of acute cholecystitis, the mRNA expression of visfatin showed the fastest increase among the other pro-inflammatory mediators studied, including interleukin (IL)-10, tumor necrosis factor-, IL-6, intracellular adhesion molecule-1, and ascular cell adhesion molecule-1. Inhibition of visfatin using siRNA abrogated the inhibitory effects of lipopolysaccharide (LPS) on the expression of ABCG1 in GBECs, suggesting that visfatin is significantly involved in the LPS-driven suppression of ABCG1. @*Conclusion@#Taken together, we concluded that visfatin is a pro-inflammatory mediators that is upregulated during acute cholecystitis and is expected to be increased within a short time after inflammation. Therefore, measuring the serum level of visfatin would be helpful in predicting the inflammatory severity in the patients with acute cholecystitis.
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BACKGROUND: Secretome refers to the total set of molecules secreted or surface-shed by stem cells. The limitations of stem cell research have led numerous investigators to turn their attention to the use of secretome instead of stem cells. In this study, we intended to reinforce antifibrotic properties of the secretome released from adipose-derived stem cells (ASCs) transfected with miR-214. METHODS: We generated miR-214-transfected ASCs, and extracted the secretome (miR214-secretome) from conditioned media of the transfected ASCs through a series of ultrafiltrations. Subsequently, we intravenously injected the miR-214-secretome into mice with liver fibrosis, and determined the effects of miR-214-secretome on liver fibrosis. RESULTS: Compared with that by naïve secretome, liver fibrosis was ameliorated by intravenous infusion of miR-214-secretome into mice with liver fibrosis, which was demonstrated by significantly lower expression of fibrosis-related markers (alpha-smooth muscle actin, transforming growth factor-β, and metalloproteinases-2) in the livers as well as lower fibrotic scores in the special stained livers compared with naïve secretome. The infusion of miR-214-secretome also led to lesser local and systemic inflammation, higher expression of an antioxidant enzyme (superoxide dismutase), and higher liver proliferative and synthetic function. CONCLUSION: MicroRNA-214 transfection stimulates ASCs to release the secretome with higher antifibrotic and anti-inflammatory properties. miR-214-secretome is thus expected to be one of the prominent ways of overcoming liver fibrosis, if further studies consistently validate its safety and efficiency.
Subject(s)
Animals , Humans , Mice , Actins , Culture Media, Conditioned , Inflammation , Infusions, Intravenous , Liver , Liver Cirrhosis , Mesenchymal Stem Cells , MicroRNAs , Research Personnel , Stem Cell Research , Stem Cells , TransfectionABSTRACT
PURPOSE: Almost all liver diseases are known to be accompanied by increased levels of reactive oxygen species (ROS), regardless of the cause of the liver disorder. However, little is known about the role of hypoxic conditioned media (HCM) in the view of pro-oxidative/antioxidative balance. METHODS: Normoxic conditioned media (NCM) and HCM were obtained after culturing adipose-derived stem cells in 20% O₂ or 1% O₂ for 24 hours, respectively. Their effects on the expression of various markers reflecting pro-oxidative/antioxidative balance were investigated in both in vitro (thioacetamide-treated AML12 cells) and in vivo (partially hepatectomized mice) models of liver injury, respectively. RESULTS: HCM treatment induced the higher expression of antioxidant enzymes, such as superoxide dismutase, glutathione peroxidase, and catalase than did NCM in the in vitro model of liver injury. We also found that HCM increased the expression of nuclear factor erythroid 2-related factor (NRF2). The in vivo models of liver injury consistently validated the phenomenon of upregulated expression of antioxidant enzymes by HCM. CONCLUSION: We thus could conclude that HCM provides protection against ROS-related toxicity by increasing the expression of antioxidant enzymes, in part by releasing NRF2 in the injured liver.
Subject(s)
Antioxidants , Catalase , Culture Media, Conditioned , Glutathione Peroxidase , In Vitro Techniques , Liver , Liver Diseases , Mesenchymal Stem Cells , Reactive Oxygen Species , Stem Cells , Superoxide DismutaseABSTRACT
PURPOSE: Everolimus only inhibits mammalian target of rapamycin complex 1 (mTORC1), whereas Ku0063794 inhibits both mTORC1 and mTORC2. Although they have similar anticancer effects, their combination has a synergistic effect against hepatocellular carcinoma (HCC) cells. We aimed to determine the mechanism underlying the synergistic effects of everolimus and Ku0063794 associated with autophagy in HCC cells. MATERIALS AND METHODS: We compared the effects of everolimus and Ku0063794, individually or in combination, on both the in vitro and in vivo models of HCCs. RESULTS: HepG2 cells treated with both agents had significantly lower rates of cell proliferation and higher apoptosis than the individual monotherapies (p < 0.05). Autophagic studies consistently indicated that, unlike the monotherapies, the combination therapy significantly reduced autophagy (p < 0.05). Autophagic blockage directly promoted the pro-apoptotic effects of combination therapy, suggesting autophagy as the survival mechanism of HCC cells. Unlike the monotherapies, combination therapy showed the potential to inhibit sirtuin 1 (SIRT1), the positive regulator of autophagy. SIRT1 overexpression abrogated the autophagy-inhibiting and pro-apoptotic effects of combination therapy. In a nude mouse xenograft model, the shrinkage of tumors was more prominent in mice treated with combination therapy than in mice treated with the respective monotherapies (p < 0.05). The immunohistochemical and immunofluorescence stains of the tumor obtained from the xenograft model showed that combination therapy had the potential of reducing autophagy and promoting apoptosis. CONCLUSION: The combination of everolimus and Ku0063794 potentiates anticancer effects on HCCs through a decrease in autophagy, which is prompted by SIRT1 downregulation.
Subject(s)
Animals , Mice , Apoptosis , Autophagy , Carcinoma, Hepatocellular , Cell Proliferation , Coloring Agents , Down-Regulation , Everolimus , Fluorescent Antibody Technique , Hep G2 Cells , Heterografts , In Vitro Techniques , Mice, Nude , Sirolimus , Sirtuin 1 , TOR Serine-Threonine KinasesABSTRACT
BACKGROUND/AIMS: Hepatitis B viral protein X (HBx) is implicated in the pathogenesis of hepatocellular carcinoma (HCC) as well as the elevation of heat shock proteins (HSPs) after hepatitis B virus (HBV) infection. We thus investigated the anticancer effects of an HSP90 inhibitor 17-Dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG) in HBx-transfected hepatocellular carcinoma cells. METHODS: pcDNA-HBx was made by inserting the HBx gene derived from the HBV-infected patient into pcDNA3.1 using the restriction enzymes (XbaI/HindIII). HBx-expressing HepG2 cells were then generated by transfecting HepG2 cells with pcDNA containing HBx gene. To compare the anticancer effects of 17-DMAG between pcDNA-HBx transfected HepG2 cells and the control cells (pcDNA-transfected HepG2 cells), we performed various molecular studies, including Ez-cytox proliferation assay, Western blot analysis, and flow cytometry. RESULTS: 17-DMAG inhibited the proliferation of pcDNA-HBx transfected HepG2 cells better than control cells (P<0.05). After treating with a various concentration of 17-DMAG (50–1,000 nM), pcDNA-HBx transfected HepG2 cells exhibited higher expression of pro-apoptotic proteins (c-caspase-3, c-caspase-8, and c-caspase-9) than did control cells (P<0.05). pcDNA-HBx transfected HepG2 cells showed higher activities of caspase-3, caspase-8, and caspase-9 than did control cells (P<0.05). Finally, we found that the expression of pro-apoptotic proteins (PARP and c-caspase-3) was considerably decreased by the use of a caspase inhibitor suggesting that 17-DMAG induces the cell death of HepG2 cells caspase-dependently. CONCLUSIONS: Our study strongly suggests that 17-DMAG has antiviral effects against HBV as well as anticancer effects against HepG2 cells. Thus, the application of 17-DMAG appears to be particularly advantageous to the HCC patients related with HBV infection.
Subject(s)
Humans , Apoptosis , Apoptosis Regulatory Proteins , Blotting, Western , Carcinoma, Hepatocellular , Caspase 3 , Caspase 8 , Caspase 9 , Caspases , Cell Death , Flow Cytometry , Heat-Shock Proteins , Hep G2 Cells , Hepatitis B virus , Hepatitis B , Hepatitis , TransfectionABSTRACT
BACKGROUND: The stem cell-derived secretome has received considerable attention as an alternative to stem cells for therapeutic applications. However, establishing optimal culture conditions is key to obtaining appropriate secretome contents. Here, the optimal culturing environment for achieving a high-efficiency secretome was determined via hypoxic preconditioning of human adipose-derived stem cells (ASC). METHODS: Normoxic conditioned media (NCM) and hypoxic conditioned media (HCM) were obtained after culturing human ASCs under normoxia (20% O2) or hypoxia (1% O2), respectively. Subsequently, both normal and thioacetamide-induced hepatotoxic hepatocytes were treated with NCM or HCM. In addition, partially hepatectomized mice were infused with control saline, NCM, and HCM. The effects on liver regeneration and serum transaminases levels were then compared. RESULTS: Hypoxic preconditioning significantly increased mRNA expression of proinflammatory cytokines (interleukin-6 and tumor necrosis factor-α) and growth factors (hepatocyte growth factor and vascular endothelial growth factor). In both normal and thioacetamide-induced hepatotoxic hepatocyte (alpha mouse liver 12 [AML12]) cell lines, HCM treatment resulted in the highest cell viability (122% and 95%, respectively), followed by NCM (111% and 78%, respectively). In addition, intravenous administration of HCM to partially hepatectomized mice resulted in substantially enhanced liver regeneration compared with the NCM group (P<0.05). CONCLUSIONS: Taken together, the secretome obtained from ASC with hypoxic preconditioning showed potential to alleviate liver damage both in vitro and in vivo. Hypoxic culture of ASC is expected to play an important role in regenerative medicine by inducing secretome production that is beneficial for improving liver regeneration.
Subject(s)
Animals , Humans , Mice , Administration, Intravenous , Hypoxia , Cell Line , Cell Survival , Culture Media, Conditioned , Cytokines , Hepatocytes , Intercellular Signaling Peptides and Proteins , Liver Regeneration , Liver , Necrosis , Regenerative Medicine , RNA, Messenger , Stem Cells , TransaminasesABSTRACT
BACKGROUND: Hemophilia B is an inheritable X-linked bleeding disorder that occurs as a consequence of genetic alterations within the factor IX (IX) gene. In the present study, pseudotyped HIV-I-derived lentiviral vectors expressing human IX (lentivirus-IX) were assessed for the ability to produce an active human IX in the animals transduced with lentivirus-IX. METHODS: The IX concentrations and activated partial thromboplastin times (aPTT) were measured from the supernatants of HeLa cells that were transduced with lentivirus-IX. In an animal study, we injected 1microgram of lentivirus-IX into the hind limbs of Sparague-Dawley (SD) rats. The IX concentrations were measured from the plasma of the vehicle injected rats and the plasma of the lentivirus-IX injected rats for 8 weeks. RESULTS: The in vitro expression of human IX was detected in a dose-dependent manner following the transduction of lentivirus-IX into the HeLa cells (control: 10+/-3 vs. 100 ng of lentivirus-IX: 1486+/-50 ng/mL, P<0.05). The aPTT also showed the tendency of dose-dependent decrease (control: 83.9+/-0.5 vs. 50 ng of lentivirus-IX: 80.1+/- 0.8 sec), but this was not statistically significant. In the animal experiment, the plasma IX concentration from the lentivirus-IX transduced rats (n=3) was significantly increased compared to the vehicle-injected rats (n=4) (5.9+/-3.9 vs. 46.4+/-20.6 ng/mL) at post-injection 1 week. CONCLUSION: This study demonstrated that in vivo delivery of lentiviral vectors expressing human IX to the muscle cells has the potential to be a therapeutic modality for hemophilia B.
Subject(s)
Animals , Humans , Rats , Animal Experimentation , Blood Coagulation Factors , Extremities , Factor IX , Genetic Therapy , HeLa Cells , Hemophilia A , Hemophilia B , Hemorrhage , Lentivirus , Muscle Cells , Plasma , ThromboplastinABSTRACT
PURPOSE: This study was designed to explore what experiences nurses had while caring and providing emotional support for patients. METHODS: Participants were eight nurses working at hospitals for more than one year. Data were collected from June, 2006 to January, 2007 through in-depth interview by using tape-recordings. Data were analysed with the phenomenological method proposed by Colazzi(1978). RESULTS: From significant statements, 4 clustered themes, 7 themes and 23 sub-themes were extracted from the essential meaning of the emotional experience of hospital nurses. The 4 clustered themes were 'movement of mind', 'affection and service for patients', 'worthwhile and conflict' and 'control oneself'. The 7 themes were 'special feeling', 'rapport formation', 'consideration', 'human interaction', 'value discovery', 'loss of volition', and 'keep to balance'. CONCLUSION: Although nurses had tough experiences for providing care for patients' emotional support, they had also experienced spiritual maturity from its experience. The result of this study would contribute for nurses not only to care for patients who need emotional support but also to develop knowledge in nursing.
Subject(s)
HumansABSTRACT
Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide), the major pungent ingredient of red pepper, has been reported to possess anti-carcinogenic and anti-mutagenic activities. In this study, the anti-migration activity of capsaicin on highly metastatic B16-F10 melanoma cells was investigated. Capsaicin significantly inhibited the migration of melanoma cells without showing obvious cellular cytotoxicity at low doses. This effect correlated with the down-regulation of phosphatidylinositol 3-kinase (PI3-K) and its downstream target, Akt. Although B16-F10 cell migration was increased by the PI3-K activator through the activation of Akt, these PI3-K activator-induced phenomena were attenuated by capsaicin. Moreover, capsaicin was found to significantly inhibit Rac1 activity in a pull-down assay. These results demonstrate that capsaicin inhibits the migration of B16-F10 cells through the inhibition of the PI3-K/Akt/Rac1 signal pathway. The present investigation suggests that capsaicin targets PI3-K/Akt/ Rac1-mediated cellular events in B16-F10 melanoma cells. Consequently, capsaicin administration should be considered an effective approach for the suppression of invasion and metastasis in malignant melanoma chemotherapy.
Subject(s)
Animals , Mice , Phosphatidylinositol 3-Kinase/metabolism , Capsaicin/pharmacology , Cell Line, Tumor , Cell Movement/drug effects , Cell Survival/drug effects , Dose-Response Relationship, Drug , Immunoblotting , Melanoma, Experimental/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction/drug effects , rac1 GTP-Binding Protein/metabolismABSTRACT
PURPOSE: To better understand the extent to which chemokines participate in the mucosal inflammatory response in patients with ulcerative colitis (UC), we assessed the expression of an array of chemokines in the colonic mucosa of UC patients. METHODS: Colonic mucosal biopsy specimens were obtained from 15 patients with UC and 12 normal controls. Messenger RNA (mRNA) levels for 10 chemokines were quantitated by reverse-transcription PCR using synthetic standard RNAs. The biopsy specimens were also cultured, and secreted chemokines in culture supernatants were assayed by ELISA. RESULTS: The mRNA expression of C-X-C (IL-8, GROalpha, GRObeta, GROgamma, ENA-78, and IP-10) and C-C (MCP-1, MIP-1beta, and RANTES), but not C (lymphotactin) chemokines was significantly higher in the affected mucosa of UC patients than in the unaffected mucosa of UC patients or in the normal mucosa of normal controls. The degree of increased expression was more prominent in the C-X-C than in the C-C chemokines. Further, the secretion of IL-8, GROalpha, ENA-78, and MCP-1 was higher in UC patients than in normal controls. Secretions of MIP-1beta and RANTES also showed a trend toward an increase in UC, but it did not reach statistical significance. CONCLUSION: The increased expression of a variety of chemokines in UC suggest that chemokines may play an important role in the immunopathogenesis of UC.
Subject(s)
Humans , Biopsy , Chemokine CCL4 , Chemokine CCL5 , Chemokines , Chemokines, CC , Colitis, Ulcerative , Colon , Enzyme-Linked Immunosorbent Assay , Interleukin-8 , Mucous Membrane , Polymerase Chain Reaction , RNA , RNA, Messenger , UlcerABSTRACT
Human colon epithelial cells secrete an array of proinflammatory cytokines that includes IL-8, MCP-1, GM-CSF, TNF alpha and IL-6. This response may serve to attract neutrophils and macrophags to the site of infection. In addition to IL-8 and MCP-1, the chemokine family contains other members, which, alone or in combination, can recruit and/or activate inflammatory and lymphoid cells. In this study, we asked whether colon epithelial cells express a broader array of chemokines than previously described. The colon epithelial cell line, Caco-2, was stimulated for 3h with IL-1 alpha, or was infected with Salmonella dublin. RNA was extracted and chemokine mRNA levels were determined by quantitative reverse transcription-PCR using internal RNA standards. Ex pression of GRO alpha, GRO beta, GRO gamma and IP-10 increased by bacterial infection or IL-l alpha stimulation. These data strongly support the notion that epithelal cells are an important and integral component of the host's natural immune system.
Subject(s)
Humans , Bacterial Infections , Chemokines , Colon , Cytokines , Epithelial Cells , Granulocyte-Macrophage Colony-Stimulating Factor , Immune System , Interleukin-1alpha , Interleukin-6 , Interleukin-8 , Lymphocytes , Neutrophils , RNA , RNA, Messenger , SalmonellaABSTRACT
PURPOSE: Chemokines are potent regulators of the host inflammatory or immune responses. Mucosal synthesis of chemokines may be important in the pathogenesis of mucosal inflammation in ulcerative colitis (UC). We performed this study to investigate the expression of C-X-C chemokine genes in UC. METHODS: Mucosal tissues were obtained from six normal controls and six UC patients by endoscopic biopsies. In patients with UC, mucosal tissues were separately obtained from both involved and uninvolved regions. RNA was extracted and mRNA levels of five C-X-C chemokines were determined by quantitative reverse transcription-PCR using internal RNA standards. RESULTS: Mucosal mRNA levels of all chemokines tested increased in the involved region of UC compared with the uninvolved region of UC or normal controls. CONCLUSION: Our data suggest that mucosal expression of C-X-C chemokines contributes to the pathogenesis of UC
Subject(s)
Humans , Biopsy , Chemokines , Chemokines, CXC , Colitis, Ulcerative , Colon , Inflammation , Mucous Membrane , RNA , RNA, Messenger , UlcerABSTRACT
Two endocrine active pituitary microadenomas were treated by transsphenoidal microsurgery and evaluated endocrinologically by radioimmunoassay of hormones. They showed significant rise of serum human growth hormone(HGH) after thyrotrophine releasing hormone(TRH) administration and no suppression by oral glucose tolerance test(GTT) before operation. Pathologic findings are acidophilic adenomas.