tretinoin and phorbolol-myristate-acetate

Matrix metalloproteinase (MMP)-21 and MMP-26 (matrilysin-2) are two recently cloned epithelial metalloproteases. Here we examined their expression in various benign skin disorders, in which macrophages and fibroblasts have been implicated as well as in cultures of these cells. Expression of MMP-21 was detected by immunohistochemistry in a subset of macrophages of granulomatous skin lesions and in fibroblasts in dermatofibromas. MMP-21 mRNA was found in THP-1, U937, HEL 299 and Hs68 cells. Furthermore, MMP-21 protein was detected by immunohistochemistry in cultures of the same cell lines. In culture MMP-21 was upregulated by phorbol myristate acetate in THP-1 cells and by retinoic acid (RA) in U937 cells, and downregulated by transforming growth factor beta 1 (TGF-beta1) in HEL 299 as assessed by Taqman quantitative polymerase chain reaction (PCR). Expression of MMP-26 was detected by immunohistochemistry in granulomatous skin diseases and actinic elastosis. MMP-26 at both mRNA and protein levels was only found in HEL 299 cells. In culture it was downregulated by TGF-beta1, RA and IL-1beta as assessed by Taqman quantitative PCR. Our results suggest these two novel MMPs are not only associated with cancer but may be important in connective tissue remodelling and pathobiology of various benign skin disorders.

Topics: Cell Line; Fibroblasts; Gene Expression Regulation; Humans; Immunohistochemistry; Interleukin-1beta; Keratolytic Agents; Macrophages; Matrix Metalloproteinases, Secreted; Monocytes; RNA, Messenger; Skin Diseases; Tetradecanoylphorbol Acetate; Transforming Growth Factor beta1; Tretinoin

PMA (10, 20 ng/ml) and doxorubicin (5-20 ng/ml) decreased the viability and MTT-activity of NALM-1 pre-B leukemic cells (3 days' treatment). Further, CD10 was downregulated, suggesting that PMA and doxorubicin induced differentiation of NALM-1 cells. However, PMA did not alter expression of B cell markers CD20 and of mIgM. In contrast to PMA, another differentiation agent ATRA did not alter CD10 expression on NALM-1 cells but affected viability after 6 days (5, 10 ng/ml). The data in this study are the first evidence that PMA and doxorubicin inhibited viability and MTT activity and induced partial differentiation, by decreasing CD10 on NALM-1 cells.

Topics: Antibiotics, Antineoplastic; Antigens, CD20; Biomarkers, Tumor; Cell Line, Tumor; Cell Survival; Dose-Response Relationship, Drug; Doxorubicin; Enzyme-Linked Immunosorbent Assay; Formazans; Humans; Immunoglobulin mu-Chains; Leukemia; Neprilysin; Syndecan-1; Tetradecanoylphorbol Acetate; Tetrazolium Salts; Time Factors; Tretinoin

Extracellular nucleotides exert a variety of biological actions through several kinds of P2 receptors in many tissues and cell types. We found that treatment with nucleotides increases intracellular Ca2+ concentration ([Ca2+]i) in SK-N-BE(2)C human neuroblastoma cells with a following order of potency: UDP > UTP > ADP >> ATP. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that specific mRNAs coding for human P2Y1, P2Y4, and P2Y6 receptors were expressed in the cells, but Northern blot analysis revealed that P2Y6 receptors were the predominant type. Activation of protein kinase C-alpha by treatment with 1 micro m phorbol 12-myristate 13-acetate dramatically inhibited both the UDP-induced [Ca2+]i rise and inositol 1,4,5-trisphosphate (IP3) generation, whereas incubation with pertussis toxin had little effect on the responses. The UDP-induced [Ca2+]i rise and IP3 production were maintained up to 30 min after stimulation, while bradykinin-induced responses rapidly decreased to the basal level within 5 min of stimulation. Pretreatment of cells with the maximal effective concentration of UDP reduced the subsequent carbachol- or bradykinin-induced [Ca2+]i rise without inhibition of IP3 generation. Neuronal differentiation of the cells by treatment with retinoic acid for 7 days did not change the expression level of P2Y6 receptors. Taken together, the data indicate that P2Y6 receptors highly responsive to diphosphonucleotide UDP are endogenously expressed in the human neuroblastoma SK-N-BE(2)C cells and that they are involved in the modulation of other phospholipase C-coupled receptor-mediated Ca2+ mobilization by depleting the IP3-sensitive Ca2+ stores.

Topics: Bradykinin; Calcium; Cell Differentiation; Cholinergic Agonists; Enzyme Activation; Humans; Inositol 1,4,5-Trisphosphate; Neuroblastoma; Nucleotides; Pertussis Toxin; Protein Kinase C; Protein Kinase C-alpha; Receptors, Purinergic P2; Receptors, Purinergic P2Y1; RNA, Messenger; Signal Transduction; Tetradecanoylphorbol Acetate; Tretinoin; Tumor Cells, Cultured; Uridine Diphosphate; Uridine Triphosphate

The conformational changes of HL60 cells during differentiation and apoptosis are still not clearly understood. This study uses a new method, Fourier-transform infrared (FT-IR) spectroscopy, to analyse the changes of HL60 cells. We detected several differences among normal HL60 cells, differentiated cells induced by all-trans retinoic acid (ATRA) and PMA, and apoptotic cells induced by vincristine (VCR). The results suggest that the alpha-helix content of the membrane protein of differentiated HL60 cells induced by ATRA and PMA increased,and that the beta-sheet content of membrane proteins of apoptotic cells induced by VCR also increased. Also, C-O(H)-stretching modes of serine, threonine and tyrosine residues of cell proteins were shown to be distinctly different, but no significant differences were detected between cells differentiated along different lineages by ATRA and PMA.

Topics: Antineoplastic Agents; Antineoplastic Agents, Phytogenic; Apoptosis; Cell Differentiation; DNA, Neoplasm; Glycoproteins; HL-60 Cells; Humans; Lipid Metabolism; Membrane Proteins; Protein Conformation; Spectroscopy, Fourier Transform Infrared; Tetradecanoylphorbol Acetate; Tretinoin; Tumor Cells, Cultured; Vincristine