Background: Despite the availability of various therapeutic strategies, the prognosis for patients with metastatic breast cancer remains poor. Epithelial-mesenchymal transition (EMT) is a critical mechanism driving metastasis in breast cancer, enabling tumor cells to lose epithelial characteristics and acquire enhanced motility and invasiveness. Results: This study investigates the role of 11alpha-hydroxyprogesterone (11α-OHP), a steroid hormone with an incompletely understood biosynthesis and metabolic pathway, in regulating lung metastasis in breast cancer. Using the MMTV-PyMT FVB mouse model, which spontaneously develops breast tumors we administered 11α-OHP for five weeks starting at 10 weeks of age. At 15 weeks, histological analysis revealed a significant reduction in lung metastasis in 11α-OHP-treated mice compared to controls, with notably smaller metastatic tumor areas in the lungs. Additionally, treated mice exhibited increased expression of epithelial cell adhesion proteins and decreased levels of focal adhesion kinase (FAK) in lung tissues. In vitro experiments using MDA-MB-231 cells corroborated these findings, showing that 11α-OHP significantly inhibited cell motility and invasiveness in scratch wound, transwell migration, and invasion assays. Notably, 11α-OHP did not significantly alter primary tumor growth in the MMTV-PyMT model. Conclusions These findings suggest that 11α-OHP may suppress breast cancer metastasis by modulating EMT, highlighting its potential as a therapeutic target for preventing metastatic progression.

Purpose: We developed immune checkpoint molecules to target recombinant dendritic cells (DCs) and verified their anti-tumor efficacy and immune response against prostate cancer. Materials and Methods: DCs were generated from mononuclear cells in the tibia and femur bone marrow of mice. We knocked down the programmed death ligand 1 (PD-L1) on monocyte-derived DCs through siRNA PD-L1. Cell surface antigens were immune fluorescently stained through flow cytometry to analyze cultured cell phenotypes. Furthermore, we evaluated the efficacy of monocyte-derived DCs and recombinant DCs in a prostate cancer mouse model with subcutaneous TRAMP-C1 cells. Lastly, DC-induced mixed lymphocyte and lymphocyte-only proliferations were compared to determine cultured DCs’ function. Results: Compared to the control group, siRNA PD-L1 therapeutic DC-treated mice exhibited significantly inhibited tumor volume and increased tumor cell apoptosis. Remarkably, this treatment substantially augmented interferon-gamma and interleukin-2 production by stimulating T-cells in an allogeneic mixed lymphocyte reaction. Moreover, we demonstrated that PD-L1 gene silencing improved cell proliferation and cytokine production. Conclusions We developed monocyte-derived DCs transfected with PD-L1 siRNA from mouse bone marrow. Our study highlights that PD-L1 inhibition in DCs increases antigen-specific immune responses, corroborating previous immunotherapy methodology findings regarding castration-resistant prostate cancer.

PURPOSE: Studies on the incidence and mortality of refractures after primary osteoporotic fracture are limited by the relatively rare incidence of such refractures and small sample sizes. The objectives of this research were: 1) to determine the incidence of osteoporotic refractures and fracture locations and 2) to assess mortality rates associated with osteoporotic refracture over a median follow up of 3 years using nationwide claim database. MATERIALS AND METHODS: Patients over 50 years of age who had an osteoporotic fracture that was confirmed operationally were enrolled. Refracture was defined as that after 6 months of an untreated period. Mortality rate was calculated using the Charlson comorbidity index and was analyzed using Cox proportional hazards regression analysis. RESULTS: A total of 18956 first-time instances of osteoporotic fracture were reported between 2007 and 2012 after a median follow up of 3.1 years (range, 1 to 7 years). Among 18956 patients, 2941 (15.50%) experienced refracture. After follow up for 1 year, cumulative mortality rates for re-fracture and non-refracture groups were 9.1% and 7.2%, respectively. After adjusting for covriates, mortality rate was 1.2 times greater in patients with re-fracture than in patients without re-fracture over a median follow up of 3 years (hazard ratio: 1.20, 95% confidence interval: 1.08–1.34, p<0.001). CONCLUSION: The incidence of osteoporotic re-fracture in this nationwide study was 15.5%, and the mortality rate of re-fracture patients was 1.2 times higher than that of non-refracture patients over a median follow up of 3 years.
Comorbidity ; Follow-Up Studies ; Humans ; Incidence ; Korea ; Mortality ; Osteoporotic Fractures ; Sample Size

PURPOSE: Biliary cancer is a highly malignant neoplasm with poor prognosis and most patients need to undergo palliative chemotherapy, however major clinical problem associated with the use of chemotherapy is chemoresistance. So far, we aimed at investigating clinical implications of apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) and Jagged1 as chemoresistance factors in biliary tract cancer. METHODS: We used 5 human biliary tract cancer cell lines (SNU-245, SNU-308, SNU-478, SNU-1079, and SNU-1196), and investigated the chemosensitivity of APEX1 and Jagged1 through 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and Western blot. Alternately, the 10 patients of advanced biliary cancer consist of 2 group according to the chemotherapy response examined by immunohistochemistry using APEX1 and Jagged1 antibody, and protein expression level was scored for staining intensity and percent positive cell. RESULTS: The result of MTT assay after APEX1 knockdown showed that strong coexpression of APEX1 and Jagged1 cell line (SNU-245, SNU-1079, and SNU-1196) showed a greater decrease in IC₅₀ of chemotherapeutic agent (5-fluorouracil, gemcitabine and cisplatin). The Western blot analysis of APEX1 and Jagged1 expression in biliary cancer cell lines after APEX1 knockdown definitively demonstrated decreased Jagged1 expression. The APEX1 and Jagged1expression level of immunohistochemistry represented that chemorefractory patients had higher than chemoresponsive patients. CONCLUSION: These results demonstrate that simultaneous high expression of APEX1 and Jagged1 is associated with chemoresistance in biliary cancer and suggest that is a potential therapeutic target for chemoresistance in advanced biliary cancer.
Biliary Tract Neoplasms* ; Biliary Tract* ; Blotting, Western ; Cell Line ; Cisplatin ; Drug Therapy ; Fluorouracil ; Humans ; Immunohistochemistry ; Prognosis

To evaluate the hepatotoxicity and nephrotoxicity of Galla Rhois (GR) toward the liver and kidney of ICR mice, alterations in related markers including body weight, organ weight, urine composition, liver pathology and kidney pathology were analyzed after oral administration of 250, 500 and 1,000 mg/kg body weight/day of gallotannin-enriched extract of GR (GEGR) for 14 days. GEGR contained 68.7+/-2.5% of gallotannin, 25.3+/-0.9% of gallic acid and 4.4+/-0.1% of methyl gallate. Also, the level of malondialdehyde (MDA), a marker of lipid peroxidation, was decreased with 19% in the serum of high dose GEGR (HGEGR)-treated mice. The body and organ weight, clinical phenotypes, urine parameters and mice mortality did not differ among GEGR-treated groups and the vehicle-treated group. Furthermore, no significant increase was observed in alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), blood urea nitrogen (BUN) and the serum creatinine (Cr) in the GEGR-treated group relative to the vehicle-treated group. Moreover, the specific pathological features induced by most toxic compounds such as CCl4 were not observed upon liver and kidney histological analysis. Overall, the results of the present study suggest that GEGR does not induce any specific toxicity in liver and kidney organs of ICR at doses of 1,000 mg/kg body weight/day, indicating that this is no observed adverse effect level (NOAEL).
Administration, Oral ; Alanine Transaminase ; Alkaline Phosphatase ; Animals ; Aspartate Aminotransferases ; Blood Urea Nitrogen ; Body Weight ; Creatinine ; Gallic Acid ; Kidney ; L-Lactate Dehydrogenase ; Lipid Peroxidation ; Liver ; Malondialdehyde ; Mice ; Mice, Inbred ICR* ; Mortality ; No-Observed-Adverse-Effect Level ; Organ Size ; Pathology ; Phenotype

Endocrine-disrupting chemicals (EDCs) are exogenous substances that alter the structure or function of the endocrine system. 4-Tert-octylphenol (OP) is one of the most representative EDCs and has estrogenic effects. In this study, we examined the effects of ethinyl estradiol (EE) and OP on the pituitary gland, placenta, and uterus of pregnant rats. Expression levels of human chorionic gonadotropin (hCG), oxytocin (OT), and contraction-associated proteins (CAPs) were determined, and uterine contractile activity was measured by uterine contraction assay. EE and OP both increased mRNA expression of OT and hCG in the pituitary gland but not the placenta. Since OT and hCG control uterine contraction, we next examined CAP expression in the uterus. Expression of 15-hydroxyprostaglandin-dehydrogenase (PGDH) was upregulated by OP, whereas expression of other CAPs was unaffected. To clarify the effect of OP on uterine contraction in pregnant rats, uterine contraction assay was performed. The 17beta-Estradiol (E2) did not affect contraction of primary uterine cells harvested from pregnant rats in a 3D collagen gel model. However, OP showed different effects from E2 by significantly reducing contraction activity. In summary, we demonstrated that OP interferes with regulation of OT and hCG in the pituitary gland as well as PGDH in the uterus, thereby reducing uterine contraction activity. This result differs from the action of endogenous E2. Collectively, these findings suggest that exposure to EDCs such as OP during pregnancycan reduce uterine contractile ability, which may result in contraction-associated adverse effects such as metratonia, bradytocia, and uterine leiomyomata.
Animals ; Chorionic Gonadotropin ; Collagen ; Endocrine System ; Estradiol ; Estrogens ; Ethinyl Estradiol ; Oxytocin* ; Pituitary Gland ; Placenta ; Rats* ; RNA, Messenger ; Uterine Contraction ; Uterus

Although formaldehyde (FA) is known to be a major allergen responsible for allergic contact dermatitis, there are conflicting reports regarding correlation between FA exposure and interleukin (IL-4) expression. To investigate whether allergic responses including IL-4 expression were induced by repeated dermal exposure to low dose FA, alterations in the luciferase signal and allergic phenotypes were measured in IL-4/Luc/CNS-1 transgenic (Tg) mice containing luciferase cDNA under control of the IL-4 promoter after exposure to 4% FA for 2 weeks. High levels of luciferase were detected in the abdominal region of the whole body and submandibular lymph node (SLN) of FA treated mice. Additionally, the ear thickness and IgE concentration were significantly upregulated in the FA treated group when compared with the acetone olive oil (AOO) treated group. FA treated mice showed enhanced auricular lymph node (ALN) weight, epidermis and dermis thickness, and infiltration of inflammatory cells. Furthermore, the expression of IL-6 among T helper 2 cytokines was higher in the FA treated group than the AOO treated group, while vascular endothelial growth factor (VEGF) levels remained constant. Overall, the results presented herein provide additional evidence that various allergic responses may be successfully induced in IL-4/Luc/CNS-1 Tg mice after exposure to low dose FA for 2 weeks. The luciferase signal under the IL-4 promoter may reflect general indicators of the allergic response induced by exposure to low dose FA.
Acetone ; Animals ; Cytokines ; Dermatitis, Allergic Contact ; Dermis ; DNA, Complementary ; Ear ; Epidermis ; Formaldehyde* ; Immunoglobulin E ; Interleukin-4 ; Interleukin-6 ; Interleukins ; Luciferases ; Lymph Nodes ; Mice ; Mice, Transgenic* ; Olea ; Phenotype ; Vascular Endothelial Growth Factor A ; Olive Oil

BACKGROUND/AIMS: Signal transducers and activators of transcription (STATs) are a family of transcription factors that are activated in response to cytokines and growth factors. STAT3 activation has been implicated in modulating the activity of downstream mediators, such as Bcl-xL and matrix metalloproteinase-2 (MMP-2). The aim of this study was to investigate the immunohistochemical expression of STAT3, B-cell lymphoma-extra large (Bcl-xL), and MMP-2 proteins according to histopathological parameters in colon adenocarcinomas, including lymph node metastasis, tumor differentiation, the TNM stage and the tumor size. METHODS: Immunohistochemical staining with monoclonal STAT3, Bcl-xL, and MMP-2 antibodies was performed on paraffin-embedded specimens from 20 colon adenomas and 39 adenocarcinomas. RESULTS: The expression of STAT3, Bcl-xL, and MMP-2 was increased in the adenocarcinomas as compared with the adenomas (p<0.001). STAT3 expression was stronger in tumors with a distant metastasis than in tumors without a distant metastasis (p=0.012). A larger tumor size was related to an increase in STAT3 expression (p=0.035). CONCLUSIONS: STAT3, Bcl-xL, and MMP-2 may play important roles in the tumorigenesis of colorectal carcinoma. STAT3 may be indicative of a poor prognosis due to its correlation with distant metastases and a larger tumor size.
Adenocarcinoma ; Adenoma ; Antibodies ; B-Lymphocytes ; Cell Transformation, Neoplastic ; Colon ; Colorectal Neoplasms ; Cytokines ; Humans ; Intercellular Signaling Peptides and Proteins ; Lymph Nodes ; Matrix Metalloproteinase 2 ; Neoplasm Metastasis ; Prognosis ; Proteins ; Transcription Factors ; Transducers

Ectonucleotide pyrophosphatase/phosphodiestrase 2 (Enpp2) isolated from the supernatant of human melanoma cells is a lysophospholipase D that transforms lysophosphatidylcholine into lysophospatidic acid. Although multiple analyses have investigated the function of Enpp2 in the hypothalamus, its role in the uterus during the estrous cycle is not well understood. In the present study, rat uterine Enpp2 was analyzed by RT-PCR, Western blotting, and immunohistochemistry. Quantitative PCR analysis demonstrated that uterine Enpp2 mRNA was decreased during estrus compared to proestrus and diestrus. To determine whether uterine Enpp2 expression is affected by sex steroid hormones, immature rats were treated with 17beta-estradiol (E2), progesterone, or both on postnatal days 14 to 16. Interestingly, the expression of Enpp2 mRNA and protein were down-regulated by E2 in the uterus during estrus but not during proestrus or diestrus, suggesting that Enpp2 may play a role in uterine function during estrus. Enpp2 is primarily localized in the stromal cells of the endometrium during proestrus and estrus. During diestrus, Enpp2 was highly expressed in the epithelial cells of the endometrium. Taken together, these results suggest that uterine Enpp2 may be regulated by E2 and plays a role in reproductive functions during female rat development.
Animals ; Estradiol/pharmacology ; Estrous Cycle/*physiology ; Female ; Gene Expression Regulation/*physiology ; Immunohistochemistry ; Mifepristone/pharmacology ; Phosphoric Diester Hydrolases/genetics/*metabolism ; Progesterone/pharmacology ; RNA, Messenger/genetics/metabolism ; Rats ; Rats, Sprague-Dawley ; Uterus/*metabolism

BACKGROUND: Limited data are available for the clinical utility of serial interferon-gamma producing T-cell response after initiation of treatment in patients with extrapulmonary tuberculosis (TB). We studied the serial TB-specific antigen T-cell responses measured using the T-SPOT.TB assay during the course of therapy. MATERIALS AND METHODS: We prospectively enrolled adult patients who were newly diagnosed with active extrapulmonary TB over a 24-month period. All patients were given standard anti-TB treatment. Blood samples were obtained for T-SPOT.TB at diagnosis, as well as 1-, 3-, 6-, and 12-months after initiating anti-TB therapy. RESULTS: A total of 52 patients with extrapulmonary TB (38 confirmed and 14 probable TB) were included in the final analysis. All patients had clinical and radiologic improvement after treatment and cured. T-SPOT.TB was positive for 90% at diagnosis, 100% at 1-, 3-, and 6-months, and 93% at 12-months after initiation of anti-TB therapy. There was no significant difference in median T-cell response between early secreting antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) at all time points. Median T-cell response steadily increased up to 6 months and then decreased. CONCLUSIONS: T-SPOT.TB assay remained positive after successful anti-TB treatment in most patients with extrapulmonary TB. Our data suggests that serial T-SPOT.TB has limited clinical utility as a surrogate marker of treatment response in patients with extrapulmonary TB.
Adult ; Biomarkers ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interferon-gamma ; Prospective Studies ; T-Lymphocytes ; Tuberculosis