No abstract available.
Fibroblast Growth Factor 7* ; Humans* ; Hyperpigmentation* ; Keratinocytes* ; Lentigo* ; Melanosis*

OBJECTIVETo detect the effects of human interleukin-1beta (IL-1beta) and Dexamethasone (DEX) on the expression level of keratinocyte growth factor (KGF) in cultured human fibroblasts of normal oral and oral lichen planus (OLP) mucosa.

METHODSThree concentration gradients of IL-1beta and DEX were added to cultured fibroblasts of human normal oral and OLP mucosa respectively. 72 hours later, the supernatant was harvested for the detection of KGF concentration with enzyme linked immunosorbent assay (ELISA). Total RNA of the fibroblasts was extracted and reverse transcribed. The resulting cDNA was then amplified by polymerase chain reaction (PCR) to detect the KGF mRNA.

RESULTSThe results of the ELISA and PCR showed that the expression levels of KGF protein and mRNA were higher if the cells were treated with IL-1beta. However, the expression levels of KGF protein and mRNA were significantly reduced if the fibroblasts were treated with DEX.

CONCLUSIONIL-1beta can promote KGF expression levels of cultured normal oral and OLP fibroblasts, and it is concentration-dependent. While DEX can inhibit KGF expression of cultured normal oral and OLP fibroblasts, and it is also concentration-dependent.

Cells, Cultured ; Dexamethasone ; Fibroblast Growth Factor 7 ; Fibroblasts ; Humans ; Interleukin-1beta ; RNA, Messenger

Sargassum fusiforme has traditionally been widely consumed in Asia as a food, and it has gained much attention due to its high nutritional, pharmaceutical, and industrial value. This study aimed to examine the promotional effects of ethanol extract (ET) and fraction obtained from ethyl acetate (FR) of S. fusiforme on hair growth in C57BL/6 mice and HaCaT cells. Five-week-old mice were used to compare hair regrowth during application of ET and FR for 21 days. Hair regrowth was evaluated by macroscopic observation and verified by hematoxylin-eosin tissue staining. Levels of mRNA expression of factors relevant to the hair growth cycle such as keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), and transforming growth factor-beta1 (TGF-beta1) were examined by quantitative polymerase chain reaction (qPCR). Our results showed that ET and FR successfully promoted hair regrowth in shaved C57BL/6 mice at a dose >20 mg/kg. Moreover, ET and FR were effective in stimulating expression of KGF and VEGF mRNAs in a dose-dependent manner, whereas TGF-beta1 was not activated. These results indicate that ET and FR of S. fusiforme effectively promoted hair growth and gene expression relevant to hair growth cycles in both in vitro and in vivo models.
Alopecia ; Animals ; Asia ; Ethanol ; Fibroblast Growth Factor 7 ; Gene Expression ; Hair* ; Mice ; Polymerase Chain Reaction ; RNA, Messenger ; Sargassum* ; Transforming Growth Factor beta1 ; Vascular Endothelial Growth Factor A

PURPOSE: Few cells with stem cell characteristics possess capabilities of self-renewal and differentiation, which leads to high tumorigenesis and resistance to standard chemotherapeutic agents. These cells are mostly quiescent, and arrest occurs at the mitotic G0/G1 phase in mitosis. We explored the effects of basic fibroblast growth factor (bFGF) on the MCF-7 cell cycle with CD44+/CD24-. METHODS: Cancer-initiating cells were propagated as mammospheres. The CD44+/CD24- subpopulation was sorted by a fluorescence activating cell sorter-Vantage flow cytometer. A cell cycle analysis was performed with different bFGF concentrations. RESULTS: Differences in the CD44+/CD24- cell proliferation under different bFGF concentrations were observed (p=0.001). When the bFGF concentration was increased, the proportion of CD44+/CD24- at G0/G1 decreased (p=0.023). CONCLUSION: We conclude that bFGF may sustain CD44+/CD24- cell proliferation and could promote cell progression through the G0/G1-->G2/S phase transition.
Breast Neoplasms ; Cell Cycle ; Cell Proliferation ; Cell Transformation, Neoplastic ; Fibroblast Growth Factor 2 ; Fibroblast Growth Factors ; Fluorescence ; MCF-7 Cells ; Mitosis ; Phase Transition ; Stem Cells

PURPOSE: The growth and PSA secretion of prostate carcinoma is predominantly regulated by androgens. However, locally produced growth factors(GFs) also have been shown to play a crucial role in the proliferation of androgen-dependent prostatic tumor cells. Androgen has been proposed to stimulate the cell proliferation and PSA secretion by modulating the activity of some of these growth factors. The objectives of this study are to determine the effects of various GFs (epidermal growth factor; EGF, basic fibroblast growth factor; bFGF, keratinocyte growth factor; KGF, hepatocyte growth factor; HGF) on the growth and PSA secretion of androgen-dependent LNCaP prostate cancer cell line. MATERIALS AND METHODS: To determine the effects of EGF, bFGF, KGF and HGF on the growth and PSA secretion of LNCaP, LNCaP cells at a concentration of 3 x 103 cells/well, suspended in T-medium containing 2% TCM, were seeded in 96 well plates. Cells were exposed to six different concentrations (0.5, 1, 5, 10, 20 and 40 ng/ml) of GFs. Cell numbers were evaluated by crystal violet assay on day 3, 5 and 7, and PSA concentrations in conditioned medium were determined on day 5. RESULTS: EGF and HGF had minimal, not significant, stimulatory effects on the growth of LNCaP. However, bFGF and KGF had significant growth stimulatory effects (P<0.05). EGF, bFGF, KGF and HGF did not have any stimulatory effects on the PSA secretion of androgen-dependent LNCaP cell line. CONCLUSIONS: bFGF and KGF, not EGF and HGF, directly stimulate the growth of LNCaP cells. However, bFGF and KGF as well as EGF and HGF do not affect the PSA secretion of LNCaP. There seems to be another signal transduction pathway, which is not associated with GFs mentioned above, for PSA secretion of androgen-dependent LNCaP cell line.
Androgens ; Cell Count ; Cell Line* ; Cell Proliferation ; Culture Media, Conditioned ; Epidermal Growth Factor ; Fibroblast Growth Factor 2 ; Fibroblast Growth Factor 7 ; Gentian Violet ; Hepatocyte Growth Factor ; Intercellular Signaling Peptides and Proteins* ; Prostate* ; Prostatic Neoplasms* ; Signal Transduction

OBJECTIVETo study the function of keratinocyte growth factor (KGF) on apoptosis of oral mucosal epithelial cells and to provide a basis for further investigation of the role of KGF in the occurrence and development of oral mucosal diseases.

METHODSDifferent concentrations of KGF (control group, 0 ng x mL(-1); experiment 1 group, 5 ng x mL(-1); experimental 2 group, 25 ng x mL(-1); experiment 3 group, 50 ng x mL(-1)) were added in oral mucosa epithelial cells cultured in vitro. After training for 12, 24, and 48 h, cell morphology was observed under an inverted microscope. Apoptosis was detected by using a flow cytometry instrument, and mRNA expression of apoptosis-related genes Bcl-2 and Bax was detected by using Real-Time fluorescent quantitative detection.

RESULTSCell adherence of the experimental group was more obvious than that of the control group, and the cell nucleolus of the experiment 3 group was obviously cultured at 48 h. After culturing for 48 h, the apoptosis rate and Bcl-2 and Bax mRNA expression among the four groups were statistically significant. The increase of KGF concentration, apoptosis rate, and Bax mRNA expression gradually reduced, whereas Bcl-2 mRNA expression increased (P < 0.05).

CONCLUSIONKGF may inhibit epithelial cell apoptosis through upregulation of Bcl-2 mRNA and downregulation of Bax mRNA.

Apoptosis ; Epithelial Cells ; Fibroblast Growth Factor 7 ; Humans ; Mouth Mucosa ; Proto-Oncogene Proteins c-bcl-2 ; RNA, Messenger

PURPOSE: In order to characterize the effects of growth factors (EGF, bFGF, KGF) on the regulation of the PSA secretion and the PSA mRNA expression of androgen- dependent LNCaP prostate cancer cell line in serum-free conditions. MATERIALS AND METHODS: LNCaP cells at a concentration of 1x104cells/well, suspended in T-medium containing 2% TCM, were seeded in 24 well plates and were exposed to four different concentrations of these growth factors to evaluate the molecular basis of PSA secretion. Cell numbers were evaluated by crystal violet assay on day 5. PSA concentrations in conditioned medium were determined on day 5, and PSA/cell number was also calculated to measure net PSA secretion per cell. PSA mRNA expression of LNCaP was assessed by RT-PCR and Northern blot analysis on day 5. RESULTS: bFGF and KGF had significant stimulatory effects (p<0.05) on the proliferation of LNCaP. However, EGF had minimal, not significant, growth stimulatory effects. EGF, bFGF and KGF did not increase the PSA secretion of LNCaP and no apparent increase or decrease in the steady-state levels of the PSA mRNA expression of LNCaP could not be detected in spite of addition of EGF, bFGF and KGF. CONCLUSIONS: bFGF and KGF, not EGF, directly stimulate the proliferation of LNCaP cells. However, bFGF and KGF as well as EGF do not affect the PSA secretion and the PSA mRNA expression of androgen-dependent LNCaP in the absence of androgenic milieu. The regulation of the PSA secretion and the PSA mRNA expression of LNCaP is not directly associated with EGF, bFGF and KGF.
Blotting, Northern ; Cell Count ; Cell Line* ; Culture Media, Conditioned ; Epidermal Growth Factor* ; Fibroblast Growth Factor 2* ; Fibroblast Growth Factor 7* ; Gentian Violet ; Intercellular Signaling Peptides and Proteins ; Keratinocytes* ; Prostate* ; Prostatic Neoplasms* ; RNA, Messenger*

OBJECTIVETo investigate the proliferative effect of keratinocyte growth factor (KGF-2) on human adult keratinocytes.

METHODSThe standard medium was keratinocyte growth medium without bovine pituitary extract (BPE), hydrocortisone or epidermal growth factor (EGF). Keratinocytes from a 48-year-old subject were cultured and seeded on dishes with standard medium of EGF in cell density of 2 x 10(4)/32 mm(2). After 24 hours, the medium was replaced by the standard medium with 0, 4, 16, 125 and 500 ng/ml KGF-2, respectively. The standard medium with EGF was used as the positive control and the standard medium without EGF or KGF-2 was used as the negative controls. The growth of keratinocytes was monitored by 3-(4,5-dimethythiazol-2-yl)-2,5 dipheyl tetrazolium bromide (MTT) assay and by photographs on days 3, 5 and 7, respectively.

RESULTSKGF-2 in concentrations of 4-500 ng/ml showed a significant proliferative effect on days 5 and 7 as compared with that of the negative controls (P < 0.01). On day 3 the cells were proliferated to 1.5-2.5-fold, on day 5 to 3-5-fold and on day 7 to 3-12-fold in KGF-2 medium as that of the negative controls. The optimal response occurred when the concentration of KGF-2 was 125 ng/ml on day 7. Cell proliferation was also consistently higher in all KGF-2 concentrations as compared with that of the positive controls.

CONCLUSIONSKGF-2 has significant effects on the proliferation of adult keratinocytes, which are more effective than that of EGF. This study supports KGF-2 can improve the healing of chronic wounds in adults in clinic.

Analysis of Variance ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Culture Media, Conditioned ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factor 7 ; Fibroblast Growth Factors ; pharmacology ; Humans ; Keratinocytes ; drug effects ; physiology ; Middle Aged ; Probability ; Reference Values ; Sensitivity and Specificity

BACKGROUNDKeratinocyte growth factor (KGF) significantly influences epithelial wound healing. The aim of this study was to isolate KGF phage model peptides from a phage display 7-mer peptide library to evaluate their effect on promoting epidermal cell proliferation.

METHODSA phage display 7-mer peptide library was screened using monoclonal anti-human KGF antibody as the target. Enzyme linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity. DNA sequencing was done to find the similarities of model peptides. Three-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, immunofluorescence assay and quantitative real-time PCR analysis were employed to evaluate the effect of the phage model peptides on epidermal cells.

RESULTSThirty-three out of fifty-eight (56.9%) of the isolated monoclonal phages exhibited high binding activity by ELISA. Ten of fifteen obtained phage model peptides were similar to KGF or epidermal growth factor (EGF). MTT assay data showed that four (No. 1 - 4) of the ten phage model peptides could promote epidermal cell proliferation. The expression of keratinocyte growth factor receptor (KGFR) mRNA in the KGF control group and the two phage model peptide groups (No. 1 and No. 2) increased. Expression of c-Fos mRNA and c-Jun mRNA in the KGF control group increased, but did not increase in the four phage model peptide groups (No.1 - 4).

CONCLUSIONFour phage model peptides isolated from the phage display 7-mer peptide library can safely promote epidermal cell proliferation without tumorigenic effect.

Cell Proliferation ; drug effects ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Epidermis ; cytology ; Fibroblast Growth Factor 7 ; chemistry ; pharmacology ; Humans ; Peptide Library ; Peptides ; chemistry ; pharmacology ; Polymerase Chain Reaction ; Receptor, Fibroblast Growth Factor, Type 2 ; genetics

BACKGROUND: The keratinocyte growth factor(KGF) is a recently identified mitogen for epithelial cells produced by nomal stromal fibroblasts. ln the skin, KGF has been shown to stimulate keratinocyte proliferation and differentiatian. Dendritic epidermal T cells(DETC) are skin-specific members of the epithelial y 8 T-cell family that reside normally in the murine epidermis. The DETCs recognize antigen expressed by damaged or diseased neighboring keratinoctyes and consequently secrete cytokines sueh as IFN- y, lL-2, IL-4, GM-CSF. OBJECTIVE: The purpose of this study was to observe the expression of KGF mRNA in keratinocyte and DETC as well as to investigate the cytokine-mediated intercellular communication between kerati- nocyte and DETC. METHODS: Using a RT-PCR(reverse transcription-polymerase chain reaction), we examined the expression of KGF mRNA in keratinocyte and DETC, and compared the level of KGF mRNA between resting and activated DETC with Con-A (concanavalin A).
Cytokines ; Epidermis ; Epithelial Cells ; Fibroblast Growth Factor 7* ; Fibroblasts ; Granulocyte-Macrophage Colony-Stimulating Factor ; Humans ; Interleukin-4 ; Keratinocytes* ; RNA, Messenger* ; Skin ; T-Lymphocytes