ABSTRACT
Intense pulsed light (IPL) devices have been shown to be highly effective for the skin rejuvenation. In our study, we try to elucidate effects of IPL in fibroblast proliferation, in gene expression, and in extracellular matrix protein production. 1BR3G human skin fibroblasts were used to test the effects of an IPL device (MiniSilk FT, Deka®). Fibroblasts were divided into three groups: group 1 was irradiated with filter 800-1200 nm (frequency 10 Hz, 15 s, fluence 60.1 J/cm) twice; group 2 was irradiated with filter 550-1200 nm (double pulse 5 ms + 5 ms, delay 10 ms, fluence 13 J/cm2) twice; and group 3 was irradiated with filter 550-1200 nm (frequency 10 Hz, 15 s, fluence 60.1 J/cm2) twice. To determine changes in gene expression, messenger RNA (mRNA) levels for collagen types I and III and metalloproteinase 1 (MMP-1) were performed 48 h after irradiation. To determine changes in hyaluronic acid, versican, and decorin, mRNA and ELISA tests were performed after 48 h of treatment. In addition to this, a Picro-Sirius red staining for collagen was made. The study showed an increase of mRNA and hyaluronic acid, decorin, and versican production. With RT-PCR assays, an increase mRNA for collagen type I, type III, and MMP-1 was observed. Collagen and hyaluronic synthesis was increased in all groups with no differences among them, while decorin and versican synthesis was higher in those groups irradiated with 550-1200-nm filters with no dependence of type pulse or total energy dose. IPL applied in vitro cultured cells increases fibroblasts activity. Synthesis of extracellular proteins seems to be produced more specifically in determined wavelengths, which could demonstrate a biochemical mechanism light depending.
Subject(s)
Collagen Type III/metabolism , Collagen Type I/metabolism , Lasers , Matrix Metalloproteinase 1/metabolism , Transcriptional Activation/radiation effects , Cell Line , Cell Proliferation , Collagen Type I/genetics , Collagen Type III/genetics , Decorin/biosynthesis , Fibroblasts/metabolism , Fibroblasts/radiation effects , Gene Expression , Humans , Hyaluronic Acid/biosynthesis , Intense Pulsed Light Therapy , Matrix Metalloproteinase 1/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Skin/cytology , Versicans/biosynthesisABSTRACT
BACKGROUND AND OBJECTIVES: Light devices emitting near infrared have been shown to be highly effective for the skin rejuvenation but biochemical and molecular mechanism or optimum dose treatment are not well-known. In our study we try to elucidate why systems emitting near infrared produce skin improvement such as fibroblasts proliferation, increase in gene expression or extracellular matrix (ECM) protein production. STUDY DESIGN/MATERIALS AND METHODS: 1BR3G human skin fibroblasts were used to test the effects of an intense pulsed light device emitting with an 800-1200 nm filter (MiniSilk FT manufactured by Deka(®)). In our protocol, fibroblasts were irradiated twice successively with a 10 Hz frequency, with a total fluence up to 60 J/cm(2) for 15s each pass. After incubating for 48 h, fibroblasts were harvested from the culture plates to test cell proliferation by flow cytometer. To determine changes in gene expression (mRNA levels for collagen types I and III and metalloproteinase 1 (MMP-1)) and protein production (hyaluronic acid, versican and decorin) tests were performed after irradiation. RESULTS: After 48 h irradiation, 1BR3G human skin fibroblasts were observed to proliferate at a fast rate. The study of ECM macromolecules production using ELISA showed an increase of hyaluronic acid and versican production but no changes were observed for decorin. With RT-PCR assays, an increase in mRNA for collagen type I, type III and MMP-1 were observed. CONCLUSION: Intense pulsed light emitting near infrared applied in vitro cultured cells increases fibroblasts proliferation and activity, which can be a possible mechanism of action for these devices in aging skin treatment.
Subject(s)
Extracellular Matrix Proteins/metabolism , Extracellular Matrix/physiology , Fibroblasts/physiology , Photic Stimulation/methods , Skin Physiological Phenomena/radiation effects , Skin/cytology , Cell Line , Cell Proliferation/physiology , Cell Proliferation/radiation effects , Dose-Response Relationship, Radiation , Extracellular Matrix/radiation effects , Fibroblasts/cytology , Fibroblasts/radiation effects , Gene Expression Regulation/physiology , Gene Expression Regulation/radiation effects , Humans , Infrared Rays , Light , Radiation Dosage , Skin/radiation effectsABSTRACT
Breast cancer metastasis suppressor-1 (BRMS1) is a putative antimetastatic gene. However, results relating its expression to the prognosis of breast cancer are still controversial, and all studies carried out to date have failed to show a relationship between the expression of BRMS1 and axillary lymph node metastasis in breast cancer. It has been recently suggested that BRMS1 may exert its physiological role through the modulation of apoptosis. In order to test this hypothesis, we studied the expression of BRMS1 and genes known to be directly related with apoptosis in human breast carcinoma. The expression of mRNA corresponding to BRMS1, BCL2, BAX, CASP3 and the apoptosis-related X-chromosome RNA binding motif (RBM) genes (RBMX, RBM3, RBM10 small and large variant) was studied by means of differential RT-PCR in 94 samples obtained from previously untreated patients with breast carcinoma. A significant (p=0.03) inverse correlation between BRMS1 mRNA expression and expression of the mRNA corresponding to the large variant of the X-chromosome RBM10 gene was found. The degree of direct correlation with another member of the X-chromosome RBM gene family, RBMX, almost attained statistical significance (p=0.06). These results point towards a possible link between BRMS1 expression and apoptosis in human breast cancer through a relationship with the expression of genes belonging to the X-chromosome RBM family.
Subject(s)
Apoptosis/genetics , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Neoplasm Proteins/genetics , Caspase 3/genetics , Chromosomes, Human, X , Female , Gene Expression Regulation, Neoplastic , Heterogeneous-Nuclear Ribonucleoproteins/genetics , Humans , Mitochondria/genetics , Mitochondria/metabolism , Proto-Oncogene Proteins c-bcl-2/genetics , RNA, Messenger , RNA-Binding Proteins/genetics , Repressor Proteins , bcl-2-Associated X Protein/geneticsABSTRACT
BACKGROUND: BRMS1, recently identified in MDA-MB-435 breast cancer cells, seems to act as a potent antimetastatic gene in experimental tumor models. MATERIALS AND METHODS: To verify if BRMS1 exerts its action in a similar way in clinical tumors, BRMS1-mRNA expression was investigated in a series of 107 human breast carcinomas and correlated with the presence or not of axillary node metastases. Additionally, BRMS1 expression was correlated with all available clinical (histologic variety, histologic and nuclear grade) and biological parameters (ploidy, expression of Ki67, hormone receptors, c-erb-B2 and p53), as well as with the expression of hMAM- and Nup88-mRNA, previously shown by us to correlate with very low and very high aggressiveness of breast cancer, respectively. RESULTS: Down-regulation of BRMS1 expression in the tumors did not correlate with the presence of axillary node metastases. Furthermore, BRMS1 expression did not correlate with any other of the studied clinical or biological tumor parameters. CONCLUSION: In clinical breast cancers, down-regulation of BRMS1 expression seems not to mimic the very clear-cut antimetatstatic properties displayed in experimental tumor models.
