Objective: To investigate the regulatory effect of the transcription factor NF-kB1 on the expression of miR-195 in prostate cancer (PCa). METHODS: We analyzed the possibility of NF-kB1 binding to the miR-195 promoter and the expression of NF-kB1 in PCa using the JASPAR and Oncomine databases, respectively, and determined the expressions of NF-kB1 and miR-195 in PCa cells by real-time quantitative PCR after inhibiting the former by interfering RNA targeting NF-kB1. We detected the activity of the luciferase reporter gene after constructing its gene plasmid in the miR-195 promoter region and having it co-transfected with the NF-kB1 plasmid. Then we analyzed the correlation between the expressions of miR-195 and NF-kB1 in the prostate tissue. RESULTS: NF-kB1 was overexpressed in PCa. After inhibition of the expression of NF-kB1, that of miR-195 was increased in PC-3 and DU-145 cell lines, with a negative correlation between the NF-kB1 and miR-195 expressions in the PCa tissue. The results of luciferase reporter gene assay showed direct binding of NF-kB1 to the miR-195 promoter zone. CONCLUSIONS NF-kB1 regulates the expression of miR-195 in prostate cancer.
Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; MicroRNAs/genetics* ; NF-kappa B p50 Subunit/metabolism* ; Promoter Regions, Genetic ; Prostatic Neoplasms/genetics* ; Transcription Factors/metabolism*

Objective: Hyperbaric oxygen treatment (HBOT) has demonstrated efficacy in improving hearing levels of patients with idiopathic sudden sensorineural hearing loss (ISSHL); however, the underlying mechanisms are not well understood. HBOT alleviates the inflammatory response, which is mediated by Toll-like receptor (TLR) 4 and nuclear factor (NF)-κB. In this study we investigated whether HBOT attenuates inflammation in ISHHL patients alteration of TLR4 and NF-κB expression. Methods: ISHHL patients ( = 120) and healthy control subjects ( = 20) were enrolled in this study. Patients were randomly divided into medicine group treated with medicine only ( = 60) and HBO group receiving both HBOT and medicine ( = 60). Audiometric testing was performed pre- and post-treatment. TLR4, NF-кB, and TNF-α expression in peripheral blood of ISSHL patients and healthy control subjects was assessed by ELISA before and after treatment. Results: TLR4, NF-κB, and TNF-α levels were upregulated in ISSHL patients relative to healthy control subjects; the levels were decreased following treatment and were lower in the HBO group than that in the medicine group post-treatment ( < 0.05 and < 0.01). Conclusion HBOT alleviates hearing loss in ISSHL patients by suppressing the inflammatory response induced by TLR4 and NF-κB signaling.
Adolescent ; Adult ; Aged ; China ; Female ; Hearing Loss, Sensorineural ; therapy ; Hearing Loss, Sudden ; therapy ; Humans ; Hyperbaric Oxygenation ; Inflammation ; genetics ; therapy ; Male ; Middle Aged ; NF-kappa B p50 Subunit ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Young Adult

Annexin A2, a multifunctional tumor associated protein, promotes nuclear factor-kappa B (NF-κB) activation by interacting with NF-κB p50 subunit and facilitating its nuclear translocation. Here we demonstrated that two ginsenosides Rg5 (G-Rg5) and Rk1 (G-Rk1), with similar structure, directly bound to Annexin A2 by molecular docking and cellular thermal shift assay. Both Rg5 and Rk1 inhibited the interaction between Annexin A2 and NF-κB p50 subunit, their translocation to nuclear and NF-κB activation. Inhibition of NF-κB by these two ginsenosides decreased the expression of inhibitor of apoptosis proteins (IAPs), leading to caspase activation and apoptosis. Over expression of K302A Annexin A2, a mutant version of Annexin A2, which fails to interact with G-Rg5 and G-Rk1, effectively reduced the NF-κB inhibitory effect and apoptosis induced by G-Rg5 and G-Rk1. In addition, the knockdown of Annexin A2 largely enhanced NF-κB activation and apoptosis induced by the two molecules, indicating that the effects of G-Rg5 and G-Rk1 on NF-κB were mainly mediated by Annexin A2. Taken together, this study for the first time demonstrated that G-Rg5 and G-Rk1 inhibit tumor cell growth by targeting Annexin A2 and NF-κB pathway, and G-Rg5 and G-Rk1 might be promising natural compounds for targeted cancer therapy.
Active Transport, Cell Nucleus ; drug effects ; Annexin A2 ; chemistry ; deficiency ; genetics ; metabolism ; Antineoplastic Agents ; chemistry ; metabolism ; pharmacology ; Apoptosis ; drug effects ; Biological Products ; chemistry ; metabolism ; pharmacology ; Cell Nucleus ; drug effects ; metabolism ; Down-Regulation ; drug effects ; Drug Discovery ; Gene Knockdown Techniques ; Ginsenosides ; chemistry ; Hep G2 Cells ; Humans ; Molecular Docking Simulation ; Molecular Targeted Therapy ; NF-kappa B p50 Subunit ; metabolism ; Protein Conformation

Chronic prostatitis is a common male disease, and its pathogenesis is not yet clear. Most scholars believe that oxidative stress and immune imbalance are the keys to the occurrence and progression of chronic prostatitis. Currently immunotherapy of chronic prostatitis remains in the exploratory stage. This article relates the active ingredients of 5 Chinese medicinal herbs (total glucosides of paeony, tripterigium wilfordii polglycosidium, curcumin, geniposide, and quercetin) for the treatment of chronic prostatitis and their possible action mechanisms as follows: 1) inhibiting the immune response and activation and proliferation of T-cells, and adjusting the proportion of Th1/Th2 cells; 2) upregulating the expression of Treg and enhancing the patient's tolerability; 3) suppressing the activation of the NF-kB factor, reducing the release of iNOS, and further decreasing the release of NO, IL-2 and other inflammatory cytokines, which contribute to the suppression of the immune response; 4) inhibiting the production of such chemokines as MCP-1 and MIP-1α in order to reduce their induction of inflammatory response. Studies on the immune mechanisms of Chinese medicinal herbs in the treatment of chronic prostatitis are clinically valuable for the development of new drugs for this disease.
Chemokines ; immunology ; Cytokines ; immunology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Immune System ; drug effects ; Male ; NF-kappa B p50 Subunit ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Plants, Medicinal ; Prostatitis ; drug therapy ; immunology ; T-Lymphocytes, Regulatory ; drug effects ; Th1-Th2 Balance

Humans ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; metabolism ; NF-kappa B p50 Subunit ; metabolism ; Transcription Factor RelA ; metabolism

Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-&kappa;B/ p65 implicates the activation of NF-&kappa;B signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-&kappa;B/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0. 5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs) , we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
Active Transport, Cell Nucleus ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Fluorescent Antibody Technique ; Human Umbilical Vein Endothelial Cells ; Humans ; NF-kappa B p50 Subunit ; metabolism ; Software

<b>OBJECTIVEb>To investigate the effect of ganoderic acid A (GA-A) on the biological behaviors of human osteosarcoma cells in vitro.

<b>METHODSb>MG63 and HOS cells were treated with 0.1, 0.25, and 0.5 mmol/L GA-A, and the changes in cell proliferation, apoptosis and migration were evaluated using MTT assay, flow cytometry, and Transwell assay, respectively. The expressions of STAT3, p38, and NF-&kappa;B1 in the cells were analyzed by Western blotting.

<b>RESULTSb>GA-A effectively inhibited the proliferation of human osteosarcoma HOS and MG-63 cells in a dose-dependent manner, and induced obvious cell apoptosis in both cells. Treatment with 0.5 mmol/L GA-A also resulted in significant inhibition of the invasion of both cells. The results of Western blotting showed that GA-A down-regulated the expression level of phosphorylated STAT3 and increased the phosphorylation level of p38 and NF-&kappa;B1 expression in both cells.

<b>CONCLUSIONb>GA-A can induce proliferation inhibition, apoptosis and suppression of invasion in human osteosarcoma HOS and MG-63 cells.

Apoptosis ; drug effects ; Bone Neoplasms ; pathology ; Cell Line, Tumor ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Heptanoic Acids ; pharmacology ; Humans ; Lanosterol ; analogs & derivatives ; pharmacology ; NF-kappa B p50 Subunit ; metabolism ; Osteosarcoma ; pathology ; Phosphorylation ; STAT3 Transcription Factor ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

<b>OBJECTIVEb>To investigate the effect of Emodin combined with 3'-azido-3'-deoxythymidine (AZT) on the proliferation and apoptosis of concentrated leukemia stem cells (CLSC)-human acute myeloid leukemia KG-la cells and expression of BCL-2, NF-&kappa;B and TGF-β.

<b>METHODSb>The tumor stem cell-like subpopulation in human leukemia cell line KG-1a was enriched with 5-fluorouracil (5-FU). The CD34⁺ CD38⁻ subpopulation in the KG-1a cells was detected with flow cytometry, the cell proliferation was detected by MTT method to study the of Emodin and AZT in the CLSC. The cell apoptosis was analyzed by flow cytometry. The expression of NF-&kappa;B, BCL-2 and TGF-β mRNA and proteins were measured with RT-PCR and Western blot respectively.

<b>RESULTSb>As compared with cells treated with mentioned above drugs alone, the inhibition of proliferation potential and apoptosis rate of cells in combination group markedly increase with time and concentration dependent member (P < 0.01), the expression of NF-&kappa;B, BCL-2 and TGF-β mRNA and proteins decreased.

<b>CONCLUSIONb>Emodin combined AZT can synergistically inhibit the proliferation, induce cell apoptosis, and down regulate the expression of NF-&kappa;B, BCL-2 and TGF-β mRNA and proteins in the CLSC, the possible mechanism of synergistic effect may be associated with inhibiton of BCL-2 activation and down-regulation of the expression of NF-&kappa;B, and TGF-β.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Emodin ; pharmacology ; Humans ; Leukemia ; NF-kappa B p50 Subunit ; metabolism ; Neoplastic Stem Cells ; cytology ; drug effects ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Zidovudine ; pharmacology

<b>OBJECTIVEb>To explore the expression of high mobility group box protein 1 (HMGB1) and nuclear factor-kappa B (NF-&kappa;B) in patients with acute leukemia and its significance.

<b>METHODb>20 samples of bone marrow and peripheral blood from each acute leukemia groups (newly diagnozed, relapsed and complete remission groups) and 20 samples as control from patients with no-hematologic malignancies were collected. The expression level of HMGB1 in peripheral blood plasma was determined by ELISA; HMGB1 and NF-&kappa;B level in mononuclear cells were examined by RT-PCR. Western blot was used to determine HMGB1 and NF-&kappa;B protein levels. HMGB1 and NF-&kappa;B in bone marrow smears were determined by immnohistochemistry method (IHC).

<b>RESULTSb>The expression level of HMGB1 obviously increased in patients of newly diagnosed and relapsed groups, as compared with control group there was statistical significance (P < 0.05), but there was no obvious difference in expression level of HMGB1 between complete remission group and control group (P > 0.05). The expression level of HMGB1 and NF-kB in monnuclear cells of bone marrow in newly-diagnosed group and relapsed group was significantly higher than that in control group (P < 0.05), but the expression levels of HMGB1 and NF-kB in complete remisson group did not change (P > 0.05). The results of immnohistochemistry method indicated that the possitive expression of HMGB1 and NF-kB maily was found in bone marrow smears of newly diagnosed and relapsed groups.

<b>CONCLUSIONb>HMGB1 is overexpressed in acute leukemia, which may be involved in the occurrence and development of acute leukemia by activating the NF-&kappa;B signaling pathway, HMGB1 may be a important index for observing therapeutic effectiveness and predicting recurrence of acute leukemia.

Acute Disease ; Blotting, Western ; Bone Marrow ; Case-Control Studies ; HMGB1 Protein ; metabolism ; Humans ; Leukemia ; diagnosis ; metabolism ; NF-kappa B p50 Subunit ; metabolism ; Remission Induction ; Signal Transduction

OBJECTIVE: To evaluate the effects of NF-κB activation on the proliferation and apoptosis throughTLR4/MyD88 signaling pathway in human nasopharyngeal carcinoma (NPC) 5-8F cell lines. METHOD: TLR4 induced by LPS is inhibited by PE anti-human. Real-Time Quantitative PCR and Western blot were employed to evaluate the efficacy of mRNA level and protein expression. The growth inhibition rate of 5-8F by Celecoxib was evaluated with MTT method. The cell cycle and apoptosis were measured with flow cytometric method (FCM). RESULT: By using the specific inhibitor, the protein and gene expression of NF-κB and MyD88 were both significantly lower than the control group (P<. 05). Meanwhile, the down-rugulation of NF-κB could inhibit proliferation of NPC 5-8F cells and promote their apoptosis (P<0. 05). CONCLUSION By inhibiting TLR4 / MyD88 signaling pathway, the expression of NF-κB in NPC 5-8F cells could decrease, then the cell proliferation was inhibited and cell apoptosis was induced. The results showed that TLR4 / MyD88 / NF-κB induced by LPS is an important pathway in the genesis and development of NPC. This study provides evidence for targeting research of NPC.
Apoptosis ; Carcinoma ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Myeloid Differentiation Factor 88 ; metabolism ; NF-kappa B p50 Subunit ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; pathology ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism