The aim of this study was to investigate the inhibitory effect of Toll-like receptor 7 (TLR7) agonist gardiquimod on K562 cells. Human γδT cells from peripheral blood cells were amplified by isopentenyl pyrophosphate. The proliferation capacity of γδT cells and K562 cells were measured with MTT assay after treatment with different concentrations of gardiquimod. Cytotoxicity of γδT cells on K562 cells was detected by CCK-8 kit, and the intracellular expression of TLR7, cell cycle and apoptosis of K562 cells before and after treatment with gardiquimod were measured by flow cytometry. The results demonstrated that gardiquimod could significantly stimulate the proliferation of γδT cells, and inhibit proliferation of K562 cells under the concentration of 11.0 µg/ml for 48 h. The expression of TLR7 increased after treatment with gardiquimod. No apoptosis was observed, but there were significant changes in cell cycle, moreover the K562 cells treated with gardiquimod were more killed by γδT cells. It is concluded that the gardiquimod can inhibit the proliferation of K562 cells and enhance their sensitivity to killing activity of human γδT cells.
Aminoquinolines ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Imidazoles ; pharmacology ; K562 Cells ; Toll-Like Receptor 7 ; agonists

OBJECTIVETo investigate the expression of Toll-like receptor 7 (TLR7) in gastric cancer cell lines and the effect of imiquimod, a TLR7 agonist, on the proliferation and apoptosis of SGC-7901 cells.

METHODSThe protein expression levels of TLR7 were detected with Western blotting in 3 human gastric cancer cell lines (SGC-7901, HGC-27 and MKN-28). The cell line expressing the highest TLR7 level was exposed to different doses of imiquimod for 12-72 h and the cell viability was assessed with MTT assay. The cell apoptosis rate after 100 µg/ml imiquimod treatment for 12 or 24 h was quantified by flow cytometry, and the ultrastructual changes of the cells were observed under electron microscope. The expression of apoptosis-related genes Bcl-2 and Bax were analyzed with real-time PCR.

RESULTSAll the 3 cell lines expressed TLR7, among which SGC-7901 cells showed the highest expression level. TLR7 agonist imiquimod dose- and time-dependently reduced the viability of SGC-7901 cells. Exposure to 100 µg/ml imiquimod for 24 h resulted in SGC-7901 cell apoptosis as shown by an increased ratio of early apoptotic cells and significant ultrastructural changes of the cells. Real-time PCR demonstrated that imiquimod treatment for 24 h caused a dose-dependent reduction of Bcl-2 mRNA expression and increment of Bax mRNA expression.

CONCLUSIONSTLR7 protein is expressed in all the 3 gastric cancer lines and its agonist imiquimod can inhibit cell proliferation and induce apoptosis in SGC-7901 cells.

Aminoquinolines ; pharmacology ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Real-Time Polymerase Chain Reaction ; Stomach Neoplasms ; metabolism ; pathology ; Toll-Like Receptor 7 ; agonists ; metabolism

cAMP-specific phosphodiesterase type 4 (PDE4) is one of the hot targets for treatment of inflammatory diseases. PDE4 inhibitors can suppress inflammation by increasing the concentration of cAMP in inflammatory cells. The efficacy and safety evaluations of several PDE4 inhibitors are currently carried on in clinical trials, for example GSK256066 in asthma, roflumilast and GSK256066 in chronic obstructive pulmonary disease, tetomilast in inflammatory bowel disease, and apremilast in dermatitis and arthritis etc. This article reviews the recent progress on PDE4-targeted therapy for inflammatory diseases.
Aminopyridines ; pharmacology ; Aminoquinolines ; pharmacology ; Arthritis ; drug therapy ; Asthma ; drug therapy ; Benzamides ; pharmacology ; Cyclopropanes ; pharmacology ; Dermatitis ; drug therapy ; Humans ; Inflammation ; drug therapy ; Inflammatory Bowel Diseases ; drug therapy ; Phosphodiesterase 4 Inhibitors ; pharmacology ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Sulfones ; pharmacology ; Thalidomide ; analogs & derivatives ; pharmacology ; Thiazoles ; pharmacology

Aminoquinolines ; pharmacology ; Anti-HIV Agents ; pharmacology ; Benzimidazoles ; pharmacology ; CCR5 Receptor Antagonists ; CD4 Antigens ; metabolism ; Cyclohexanes ; pharmacology ; HIV-1 ; pathogenicity ; physiology ; Heterocyclic Compounds ; pharmacology ; Heterocyclic Compounds, 1-Ring ; Humans ; Maraviroc ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; Receptors, CCR5 ; metabolism ; Receptors, CXCR4 ; antagonists & inhibitors ; metabolism ; T-Lymphocytes ; virology ; Triazoles ; pharmacology ; Viral Tropism

Epidermal growth factor receptor (EGFR)-targeted therapies have been effective in some cancers, but not in hepatocellular carcinoma (HCC). The aim of this study was to investigate the drug potential to overcome multi-drug resistance in HCC cells. Thirteen drug-sensitive HCC cells were assessed using the CCK-8 assay. G0-G1 arrest was measured by FACS. Western blot analysis was used to detect the key enzymes in both the Ras/Raf and PI3K pathways. When establishing the IC50 of HCC to several drugs, including EKB-569, sorafenib, erlotinib, gefitinib, pazopanib, and brivanib, SK-Hep1 cells treated with EKB-569 have shown the highest (72.8%-86.4%) G0-G1 arrest and decreased the phosphorylation of AKT and ERK at the protein level. We found that EKB-569 had higher efficacy in HCC, compared to first generation, reversible EGFR-TK inhibitors. Furthermore, the combination of sorafenib and EKB-569 showed a synergistic effect to inhibit proliferation of SNU-475, previously the most resistant cell to EGFR-TKIs. Therefore, novel EKB-569 in combination with sorafenib may be able to overcome HCC resistance to EGFR-TK inhibitors.
Aminoquinolines/*pharmacology ; Aniline Compounds/*pharmacology ; Antineoplastic Agents/*pharmacology ; Antineoplastic Combined Chemotherapy Protocols/pharmacology ; Benzenesulfonates/pharmacology ; Carcinoma, Hepatocellular/*drug therapy/pathology ; Cell Cycle Checkpoints/drug effects ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Drug Resistance, Neoplasm ; Drug Synergism ; Humans ; Liver Neoplasms/drug therapy/pathology ; Mitogen-Activated Protein Kinases/*metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt/*metabolism ; Pyridines/pharmacology ; Receptor, Epidermal Growth Factor/*antagonists & inhibitors

To explore the role of methotrexate (MTX) in regulating the number of regulatory T cells (Treg) and the mRNA expression of transcription factor Foxp3.
 Methods: 1) We analyzed the number of Treg and the mRNA expression of Foxp3 by flow cytometry (FCM) and quantitative real-time PCR (qRT-PCR) respectively in patients with psoriasis vulgaris, patients with psoriasis vulgaris after the 8-week treatment of MTX, and healthy people. 2) BALB/c female mice were smeared with imiquimod (IMQ) cream for 6 days. We recorded the change of the lesion in mice every day. The morphological changes of lesion in mice were evaluated by the psoriasis area and severity index (PASI) and HE staining. 3) The mouse model was randomly divided into a control group and an MTX group. The MTX group was treated with different doses of MTX (38.5 and 77.0 nmol/L) on the third day of this experiment. The morphological changes of lesion in mice were evaluated by PASI and HE staining. We tested the number of Treg and the expression level of Foxp3 mRNA in splenic lymphocytes.
 Results: 1) The number of Treg and the expression level of Foxp3 mRNA were lower in psoriasis vulgaris patients than those in the healthy control group (P<0.05). After 8-week treatment of MTX, the number of Treg was increased (P<0.05) and Foxp3 mRNA level was up-regulated (P<0.01). 2) Typical psoriasis-like skin lesions, such as red scaly skin plaque were found after topical application of IMQ. Both the number of Treg in the splenic lymphocytes of mice and the Foxp3 mRNA level of Treg were reduced by IMQ (P<0.01 and P<0.05). 3) Different doses of MTX for mice showed the ability to improve skin lesion, increase the number of Treg in the spleen of mice and Foxp3 mRNA level in psoriatic dermatitis of mice (P<0.05).
 Conclusion: MTX is able to regulate the number of Treg and Foxp3 mRNA expression in psoriasis.
Adjuvants, Immunologic ; pharmacology ; Aminoquinolines ; pharmacology ; Animals ; Case-Control Studies ; Female ; Forkhead Transcription Factors ; metabolism ; Humans ; Imiquimod ; Immunosuppressive Agents ; administration & dosage ; pharmacology ; Lymphocyte Count ; Methotrexate ; administration & dosage ; pharmacology ; Mice ; Mice, Inbred BALB C ; Psoriasis ; drug therapy ; immunology ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Random Allocation ; Spleen ; cytology ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; metabolism

OBJECTIVETo investigate the effects of the quinoline derivative PQ1 combined with cisplatin on the proliferation and gap junction communication of prostate cancer PC3 cells.

METHODSWe cultured in vitro prostate cancer PC3 cells and divided them into DMSO blank control, cisplatin control, and cisplatin (10 mg/ml) plus PQ1 (1, 2, 5, 10, and 15 μmol/L) groups. We measured the proliferation of the prostate cancer PC3 cells, determined the expressions of the connexin 43 (Cx43) mRNA and protein by RT-PCR and Western blot, and compared the indexes among different groups.

RESULTSCisplatin combined with PQl at 1 - 10 μmol/L significantly inhibited the proliferation of the PC3 cells and the inhibition rate rose in a concentration- and time-dependent manner, from (48.72 ± 0.98)% vs (50.33 ± 0.62)% at 0 μmol/L to (77.38 ± 1.12)% vs (83.50 ± 1.05)% at 15 μmol/L at 24 and 48 hours (P < 0.05). Compared with the cisplatin control, cisplatin combined with PQ1 at 1, 2, 5, 10, and 15 μmol/L increased the expression of Cx43 mRNA from 0.379 ± 0.113 to 0.669 ± 0.031, 0.831 ± 0. 127, 0.769 ± 0.100, 0.532 ± 0.086, and 0.475 ± 0.134, respectively (P < 0.05), and cisplatin combined with PQ1 at 1, 2, 5, and 10 μmol/L elevated that of Cx43 protein from 0.138 ± 0.146 to 0.263 ± 0.111, 0.306 ± 0.152, 0.415 ± 0.280, and 0.643 ± 0.310, respectively (P < 0.05).

CONCLUSIONThe quinoline derivative PQ1 can promote the gap junction communication of prostate cancer PC3 cells and enhance the killing effect of cisplatin on PC3 cells by upregulating the expressions of Cx43 mRNA and protein.

Aminoquinolines ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Connexin 43 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Gap Junctions ; drug effects ; physiology ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; physiopathology ; RNA, Messenger ; metabolism ; Time Factors

Aminoquinolines ; pharmacology ; therapeutic use ; Animals ; Asthma ; drug therapy ; immunology ; pathology ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Count ; Chemokine CCL17 ; Chemokine CCL22 ; Chemokines, CC ; analysis ; genetics ; Cytokines ; biosynthesis ; Flow Cytometry ; Interleukin-4 ; antagonists & inhibitors ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis ; STAT6 Transcription Factor ; analysis ; antagonists & inhibitors ; genetics ; Signal Transduction ; drug effects