Tumor-Associated Macrophage-Induced Invasion and Angiogenesis of Human Basal Cell Carcinoma Cells by Cyclooxygenase-2 Induction

Tumor-associated macrophages (TAMs) and cyclooxygenase-2 (COX-2) are associated with invasion, angiogenesis, and poor prognosis in many human cancers. However, the role of TAMs in human basal cell carcinoma (BCC) remains elusive. We found that the number of TAMs infiltrating the tumor is correlated with the depth of invasion, microvessel density, and COX-2 expression in human BCC cells. TAMs also aggregate near COX-2 expressing BCC tumor nests. We hypothesize that TAMs might activate COX-2 in BCC cells and subsequently increase their invasion and angiogenesis. TAMs are a kind of M2 macrophage derived from macrophages exposed to Th2 cytokines. M2-polarized macrophages derived from peripheral blood monocytes were cocultured with BCC cells without direct contact. Coculture with the M2 macrophages induced COX-2-dependent invasion and angiogenesis of BCC cells. Human THP-1 cell line cells, after treated with phorbol myristate acetate (PMA), differentiated to macrophages with M2 functional profiles. Coculture with PMA-treated THP-1 macrophages induced COX-2-dependent release of matrix metalloproteinase-9 and subsequent increased invasion of BCC cells. Macrophages also induced COX-2-dependent secretion of basic fibroblast growth factor and vascular endothelial growth factor-A, and increased angiogenesis in BCC cells

Outbreak of Contact Dermatitis Related to Acticide Ep Paste in a Paint Manufacturing Factory

An outbreak of severe itching, erythematous and edematous dermatitis over the extremities and upper back developed in 8 of 17 workers in the rawmaterials department of a paint manufacturing factory. The outbreak occurred during a 2- month period when Acticide EP paste (Thor Chemical, Cheshire , UK) was used in place of Metatin as a microbiocide (Acima Chemical, Buchs, Switzerland). To evaluate the frequency and the etiologic agent of this outbreak, a plant walk-through , examination and review of photographs of skin lesions followed by statistical analysis for association between the development of dermatitis and exposure to Acticide paste were performed. Three guinea pigs were subjected to patch tests comparing the dermatotoxicity of Acticide EP and Metatin. The results showed that 8 out of 17 workers (47%) suffered from contact dermatitis during the 2-month period. Stratification by occupational exposure further confirmed the association between the development of dermatitis and exposure to the Acticide paste. The dermatotoxicity test on guinea pigs revealed the marked corrosive effect of the paste and the absence of dermatotoxicity of Metatin. After the removal of the paste from the raw material, there were no newcases of contact dermatitis at the 6 month follow-up. We conclude that Acticide EP paste was the responsible offending agent. Because siothiazolinone derivatives are well-known antigens and 2-n-octy-4-isothiazolin-3-one is the active ingredient in Acticide EP paste, 2-n-octyl-4- isothiazolin-3-one is the likely cause of the dermatitis

Effects of Estrogen and Estrogen Receptor in Normal Human Melanocytes

Normal human melanocytes were cultured selectively with F12 culture medium supplemented with growth hormones, phorbol ester and 1% of fetal calf serum. The estrogen receptors were analyzed using hydroxylapatite- column assay with tritiated 17-beta-estradiol as the binding ligand. Phenol red- free medium was used when the changes in cell numbers, melanin content and tyrosinase were assessed after incubating with physiological concentration of 17-beta estradiol (10(-12) and 10(-9) M). It was found that the melanocytes contained both cytosol (5.42 +/- 1.11 fmol/mg protein ) and nuclear (59.13 +/- 17.12 fmol/mg protein) estrogen receptor. In response to estradiol, the cell number increased but both the melanin content and the tyrosinase activity decreased in a dose related pattern. These data suggested the presence of estrogen receptor with biological function in normal human melanocytes

Interleukin-6 Induced Basic Fibroblast Growth Factor-Dependent Angiogenesis in Basal Cell Carcinoma Cell Line Via Jak/Stat3 and Pi3- Kinase/Akt Pathways

We have previously demonstrated a xenograft of interleukin-6 (IL-6) overexpressing basal cell carcinoma (BCC) cell line induced tumors with high vasculature in nude mice. Here we asked whether IL-6 could induce angiogenic activity in BCC cell line. Tenfold concentrated conditioned medium (CM) from IL-6 overexpressing BCC cells exhibited higher angiogenic activities in chorioallantoic membrane and Matrigel plug assays, when compared with CM from vector control or parental BCC cells. The level of basic fibroblast growth factor 2 (bFGF) mRNA and secreted bFGF increased in IL-6 overexpressing BCC cells as shown by RT-PCR and ELISA, respectively. Concordantly, recombinant IL-6 treatment caused the elevation of bFGF mRNA and protein levels in parental BCC cells in a time- dependent manner. Neutralizing bFGF function by anti-bFGF antibody significantly inhibited CM-induced human umbilical vein endothelial cells (HUVEC) tube formation and Matrigel plug formation. Meanwhile, cyclooxygenase 2 (COX-2)-specific siRNA markedly abolish HUVEC tube formation. These data indicated both bFGF and COX -2 play an essential role for IL-6-induced angiogenesis in BCC cell line. Treatment with AG490 (Janus tyrosine kinase [ JAK] inhibitor) and LY294002 (PI3-Kinase inhibitor) inhibited IL-6-mediated upregulation of bFGF mRNA and protein secretion. Consistently, transfection with dominant negative mutants of signal transducer and activator of transcription 3 (STAT3) and acutely transforming retrovirus AKT8 in rodent T cell lymphoma (Akt) effectively abolished IL-6-mediated expression of bFGF mRNA and protein. Our data suggest that under in vitro experimental condition, bFGF and COX-2 are downstream effectors of IL-6-induced angiogenic activity in BCC cell. The IL-6-mediated bFGF upregulation is through activation of JAK/STAT3 and PI3 -Kinase/Akt pathways

Curcumin Induces a P53-Dependent Apoptosis in Human Basal Cell Carcinoma Cells

Curcumin, a potent antioxidant and chemopreventive agent, has recently been found to be capable of inducing apoptosis in human hepatoma and leukemia cells by way of an elusive mechanism. Here, we demonstrate that curcumin also induces apoptosis in human basal cell carcinoma cells in a dose- and time-dependent manner, as evidenced by internucleosomal DNA fragmentation and morphologic change. In our study, consistent with the occurrence of DNA fragmentation, nuclear p53 protein initially increased at 12 h and peaked at 48 h after curcumin treatment. Prior treatment of cells with cycloheximide or actinomycin D abolished the p53 increase and apoptosis induced by curcumin, suggesting that either de novo p53 protein synthesis or some proteins synthesis for stabilization of p53 is required for apoptosis. In electrophoretic mobility gel-shift assays, nuclear extracts of cells treated with curcumin displayed distinct patterns of binding between p53 and its consensus binding site. Supportive of these findings, p53 downstream targets, including p21(CIP1/WAF1) and Gadd45, could be induced to localize on the nucleus by curcumin with similar p53 kinetics. Moreover, we immunoprecipitated extracts from basal cell carcinoma cells with different anti-p53 antibodies, which are known to be specific for wild-type or mutant p53 protein. The results reveal that basal cell carcinoma cells contain exclusively wild-type p53; however, curcumin treatment did not interfere with cell cycling. Similarly, the apoptosis suppressor Bcl-2 and promoter Bax were not changed with the curcumin treatment. Finally, treatment of cells with p53 antisense oligonucleotide could effectively prevent curcumin-induced intracellular p 53 protein increase and apoptosis, but sense p53 oligonucleotide could not . Thus, our data suggest that the p 53-associated signaling pathway is critically involved in curcumin-mediated apoptotic cell death. This evidence also suggests that curcumin may be a potent agent for skin cancer prevention or therapy

Bipyridyl Dihydrochloride Inhibits Tumor Necrosis Factor-Alpha Secretion by Human Keratinocytes on Ultraviolet Irradiation

Paraquat manufacturers in Taiwan have been found to develop solar lentigo , actinic keratosis, as well as skin cancer in sun-exposed areas. Bipyridine has been found to be the responsible agent. At present, the mechanism for the pathogenesis of bipyridine-induced skin cancer is not known and it may be multifactorial. We investigated possible alterations in tumor necrosis factor (TNF)-alpha secretion in keratinocytes treated with 4,4'-bipyridyl dihydrochloride in vitro. Normal human keratinocytes were cultured and treated with bipyridine (10 micrograms/mL) or ultraviolet B( UVB) light (10 mJ/cm2), or with a combination of both. Bipyridine treatment alone resulted in a significant reduction in constitutive TNF-alpha secretion. Furthermore, in contrast to the finding in normal keratinocytes, UVB irradiation failed to promote TNF-alpha secretion in bipyridine-primed keratinocytes

The Phosphotidyl Inositol 3-Kinase/Akt Signal Pathway Is Involved in Interleukin-6-Mediated Mcl-1 Upregulation and Anti-Apoptosis Activity in Basal Cell Carcinoma Cells

Dysregulation of interleukin-6 has been reported to be associated with various types of tumors, and interleukin-6 plays an important part in regulating apoptosis in many types of cells. Previously, Mcl-1 was shown to be significantly increased in interleukin-6- overexpressed basal cell carcinoma cells and conferred on them anti- apoptotic activity. The aim of this study was to investigate which signaling pathway is involved in the anti-apoptotic effect of interleukin-6 on basal cell carcinoma cells. Here we show that the addition of recombinant 100 ng per ml interleukin-6 to basal cell carcinoma cells induced a 2.3- fold increase in the level of Mcl-1 protein in basal cell carcinoma cells. Transfection with dominant- negative STAT3 ( STAT3F) into interleukin-6- treated basal cell carcinoma cells caused a decrease of phosphotyrosyl STAT3 but did not alter Mcl-1 protein levels; however, AG490, a Janus tyrosine kinase inhibitor, was capable of inhibiting the interleukin -6-induced elevation of Mcl-1 protein. Next, interleukin-6 stimulation elicited extracellular signal- regulated kinase activation in basal cell carcinoma cells, and the mitogen - activated protein kinase inhibitor, PD98059, could affect this response without affecting the interleukin-6- medi-ated Mcl-1 upregulation. Use of the two phosphotidyl inositol 3- kinase inhibitors, LY294002 and wortmannin, to check whether this pathway is involved in Mcl-1 upregulation by interleukin-6, we found that the phosphotidyl inositol 3- kinase inhibitors completely attenuated the interleukin-6- induced Mcl-1 upregulation. Furthermore, in the interleukin- 6-overexpressing basal cell carcinoma cell clone, dominant- negative Akt also significantly reduced the increased level of Mcl-1. Interestingly, Janus tyrosine kinase inhibitor, AG 490, treatment strongly blocked the phosphotidyl inositol 3- kinase pathway activation, as evidenced by the decrease in phospho-Akt level. Blockage of phosphotidyl inositol 3- kinase/Akt pathway abolished the interleukin-6-mediated anti -apoptotic activity in ultraviolet B treated cells. Unexpectedly, without ultraviolet B irradiation, STAT3F transfection also induced a significant apoptosis in basal cell carcinoma/ interleukin-6 cells. Taken together, our data suggest that both the phosphotidyl inositol 3-kinase/Akt and STAT3 pathways are potentially involved in interleukin-6 -mediated cell survival activity in basal cell carcinoma cells; however, the upregulation of the anti-apoptotic Mcl-1 protein by interleukin-6 is mainly through the Janus tyrosine kinase/ phosphotidyl inositol 3-kinase/ Akt, but not the STAT3 pathway

Clinical Studies on Sporotrichosis in Taiwan

A total of 59 cases, who visited the Dermatological Clinic of National Taiwan University Hospital in the past 27 years, was proved to be Sporotrichosis. The diagnostic criteria were the suggestive clinical pictures with any one of the following points: (1) positive fungal culture with characteristic colonies of Sporotrichum schenckii, (2) good therapeutic response to potassium iodide, (3) demonstration of fungal elements in the suggested pathological tissues or( 4) demonstration of fungal elements in pus or tissue using direct immunofluorescence. All the informations concerning the clinical pictures and epidemiological points of view of these 59 patients were analysed. The chief cutaneous lesions were composed of nodules, ulcers or infiltrated plaques in linear arrangement on the exposed areas especially arms, legs or face. Thirty eight cases were male and 21 female. It affected peoples of the age between 4 to 70 years old, and their occupations were mainly outside workers such as agriculturist, worker, etc. Most of the affected people lived in the suburban areas of Taipei city such as Taoyuan and Taipei county. Serological tests such as precipitation, complement fixation and hemagglutination tests were performed in 12 cases, yet no reliable results were got