以碱性成纤维细胞生长因子及其受体为靶点的黑素瘤治疗

碱性成纤维细胞生长因子的促增殖和促血管形成的功能在黑素瘤的发生、发展和转移中起重要作用。近来关于以成纤维细胞生长因子及其受体为靶点的黑素瘤治疗的研究逐渐增多，抗肿瘤生物治疗的特异性和靶向性已成为当前肿瘤治疗学中的研究热点。     

成纤维细胞生长因子及其受体在毛发生长周期中的表达和作用

成纤维细胞生长因子(FGFs)家族各成员及其受体(FGFRs)在毛发生长周期中起重要作用。在毛发生长周期的不同阶段,FGF1、FGF2、FGF5、FGF7、FGF10、FGF13、FGF18及FGF22在毛囊中均有较高的表达,它们通过与其受体结合介导信号通路传递信息至细胞内,在毛囊生长中发挥不同的作用。该文对FGFs与FGFRs在毛发生长周期作用的研究进展作一综述。     

成纤维细胞生长因子受体在人正常皮肤和伤口处的免疫定位

成纤维细胞生长因子(FGF)是一个有22个成员的细胞因子家族，与细胞表面受体结合调节细胞生长、增殖、分化和基质合成并参与炎症反应，其作用与细胞分化及生化状态和理化环境有关。目前发现有4种成纤维细胞生长因子受体(FGFR)，作者采用免疫组化技术检测了4种受体在正常皮肤和伤口处的表达。结果：①正常皮肤中FGFR的表达：FGFR-1表达于表皮各层、皮肤附属器、竖毛肌、血管和真皮成纤维细胞；FGFR-2在表皮、毛囊、皮脂腺、小汗腺、血管和成纤维细胞有表达，而且在基底层呈强阳性；FGFR-3在基底层的上部和毛囊内层呈强阳性；FGFR-4主要见于血管平滑肌、竖毛肌和小汗腺；②头皮生长期毛囊FGFR的表达：FGFR-1在毛发基质细胞、内外根鞘和毛乳头细胞强表达；FGFR-2的表达定位与FGFR-1相似，并且在毛囊根部基质细胞呈强阳性而角化区较弱；FGFR-3在毛球至峡部的内毛根鞘内段强表达，也见于外毛根鞘峡部和已分化的皮脂腺细胞及皮脂腺导管；FGFR-4仅见于毛囊周结缔组织鞘；③伤口处FGFR的表达：新生表皮基底层细胞FGFR-1和3呈强表达，而FGFR-2和4的表达较弱；基底层上部的细胞FG．FR-3强表达，而FGFR-1和2表达弱；肉芽和瘢痕组织中成纤维细胞／肌纤维母细胞表面FGFR-1和3的表达较强，而2和4呈弱阳性，而浸润单核细胞表面FGFR-1和3表达较强。讨论：正常表皮中4种FGFR表达的变化，提示在角质形成细胞分化过程中4种受体的表达发生着改变。研究表明，表皮细胞特异性表达FGFR-3亚型Ⅲb，而间质细胞同时表达FGFR-3的两个亚型Ⅲb和Ⅲc。其中Ⅲb与aFGF和FGF-9结合，而Ⅲc与aFGF和FGF-4高亲和力结合，与bFGF结合力弱而不与FGF-5结合。bFGF位于表皮基底层而aFGF则主要存在于棘层。aFGF和FGFR-3在基底层上部的正在分化的角质形成细胞中共表达，提示aFGF可能通过自分泌作用参与角质形成细胞的分化过程。Finch等发现基质细胞表达对人角质形成细胞有强促分裂作用的角质形成细胞生长因子，并发现其受体表达于正常人表皮基底层及银屑病表皮基底层和基底层上方的角质形成细胞。本研究发现，基底层中FGFR-2的表达，可能是角质形成细胞生长因子受体在表皮基底层和外毛根鞘的定位。Gonzalez等发现FGFR-1及其mRNA主要表达于表皮生发层，由此推测bFGF可能通过FGFR-1对基底细胞有促分裂作用，而aFGF则促进棘细胞的有丝分裂。动物实验表明：FGF和FGFR的表达与毛发的生长周期和发育有关。作者发现4种FGFR在正常人皮肤生长期毛囊的表达模式不同，特别在毛球根部毛母质细胞中FGFR-2的表达呈强阳性，结合动物实验结果，本研究提示真皮乳头FGF-2活化毛母质细胞表达FGFR-2并刺激毛母质细胞增生和毛发生长；毛囊内FGFR-1的表达，提示FGFR-1可能在角质形成细胞向毛发分化的终末过程中起一定的作用。FGFR-3在生长期毛囊中内外毛根鞘和皮脂腺的终末分化细胞中表达而在毛基质细胞内不表达，这可能是由于分化类型的差异，角质形成细胞表达不同的FGFR。     

以碱性成纤维细胞生长因子及其受体为靶点的黑素瘤治疗

碱性成纤维细胞生长因子的促增殖和促血管形成的功能在黑素瘤的发生、发展和转移中起重要作用。近来关于以成纤维细胞生长因子及其受体为靶点的黑素瘤治疗的研究逐渐增多,抗肿瘤生物治疗的特异性和靶向性已成为当前肿瘤治疗学中的研究热点。     

成纤维细胞生长因子10在寻常型银屑病皮损中的检测

目的检测成纤维细胞生长因子10(FGF10)在寻常型银屑病皮损中的水平,探讨其与银屑病发病的关系。方法应用免疫组化法检测了FGF10在22例寻常型银屑病皮损、非皮损中的分布。以20例正常皮肤为正常对照。结果寻常型银屑病皮损、非皮损中FGF10的阳性检测结果均明显高于对照组(P<0.05)。结论银屑病皮损中FGF10阳性的检测结果可能与银屑病的发病有关。     

1320nm激光对成纤维细胞增殖和分泌碱性成纤维细胞生长因子及转化生长因子β1的影响

目的 探讨非剥脱性1320nm激光对体外培养的人真皮成纤维细胞增殖和分泌碱性成纤维细胞生长因子(bFGF)和转化生长因子(TGF-β1)的影响.方法 体外培养人真皮成纤维细胞,用1320nm波长激光15J/cm²,20J/cm²和24J/cm²三个能量分别照射3次.于照射后0、24、48和72 h ELISA法检测上清液bFGF和TGF-β1的分泌水平,并计算细胞总数.结果 20J/cm²以上能量激光照射后成纤维细胞计数增加,与未照射的对照组比较,差异有统计学意义(P<0.05).激光照射后bFGF分泌的量,在20J/cm²和24J/cm²能量组增加,尤以20J/cm²为著,24 h达高峰(P<0.01),与对照组比较,差异有统计学意义.TGF-β1分泌的量在照射的各能量组均下降,能量越高,下降越明显,24h达峰谷,与对照组比较,高能量组P<0.01,其他2个能量组P<0.05.结论 1320nm波长激光使成纤维细胞分裂增殖加速,并促进bFGF分泌,抑制TGF-β1分泌.     

碱性成纤维细胞生长因子治疗 CO2激光术后创面愈合疗效观察

碱性成纤维细胞生长因子（basicfibroblastgrowthfactor,bFGF）是人体内含有的一种微量活性蛋白,对源于中胚层和神经外胚层的细胞和组织,有促进分化和调节作用。我科用珠海东大生物制药有限公司研制的贝复济（主要成分为bFGF）治疗CO２激光术后溃疡,取得了满意疗效,现报道如下。一、一般资料患者２００例均来自门诊,男８７例,女１１３例。其中尖锐湿疣１２２例,面部黑痣６９例,睑黄瘤９例,年龄１５～６５岁,平均３１．１５岁,创面面积０．４cm×０．５cm～１．０cm×３．０cm,平均１．１２cm２。将患者分成观察组１２０例…     

碱性成纤维细胞生长因子对皮肤萎缩性瘢痕中成纤维细胞的作用

目的探讨人碱性成纤维细胞生长因子(bFGF)对豚鼠皮肤萎缩性瘢痕中成纤维细胞的作用。方法取实验用成年豚鼠20只,于脊柱旁两侧A,B,C三处皮肤分别人工造成萎缩性瘢痕后,实验组在A处真皮层注射50μg/L的bFGF0.1mL,隔日1次,共4次;在C处真皮层注射同剂量生理盐水作阴性对照,B处不作任何处理做空白对照。术后第21天切取瘢痕组织行病理切片,应用鼠抗人ki-67单克隆抗体行免疫组化,显微镜下计算增殖成纤维细胞所占百分率。结果实验组、阴性对照组和空白对照组切口瘢痕增殖成纤维细胞百分率分别为(7.63±1.42)%,(0.98±0.33)%和(1.22±0.34)%。实验组增殖成纤维细胞百分率与阴性对照组及空白对照组比较显著增高,差异有统计学意义(P<0.05)。结论 bFGF可以促进豚鼠皮肤萎缩性瘢痕中的成纤维细胞增殖。     

寻常型银屑病患者血清对真皮成纤维细胞生长的影响

银屑病患者体内多种细胞因子发生了变化，这些因子与临床症状之间的关系有不同报道［１］。我们研究了银屑病患者血清对成纤维细胞３Ｈ－ＴｄＲ掺入的影响，探讨掺入活性与临床症状之间的关系及体液因素在发病中的作用。一、病例和方法（一）病例：寻常型银屑病患者２１例...     

皮肤血管瘤患儿尿液中碱性成纤维细胞生长因子的检测

目的 检测尿液中碱性成纤维细胞生长因子(bFGF)水平在血管瘤病程中的变化,为临床监测血管瘤的发生发展提供客观依据.方法 采用酶联免疫吸附法(ELISA)检测27例增生期皮肤血管瘤患儿浅层X线治疗前后尿液中bFGF水平的变化,同时选择30例健康婴幼儿尿液作为对照.结果 增生期血管瘤患儿治疗前尿液中bFGF水平高于治疗后3个月和对照组,差异有统计学意义(t=0.206,P＜0.05),治疗后3个月与对照组之间bFGF水平的差异无统计学意义(t=0.505,P＞0.05).结论 ELISA检测尿液中bFGF方法简便、可靠,可为临床监测血管瘤发生发展提供客观指标.

Abstract：

Objective To determine the changes of urine bFGF level in the progression of cutaneous hemangioma,and to provide an objective index for the monitoring of hemangioma occurrence and development.Methods Enzyme-linked immunosorbent assay (ELISA) was used to determine the bFGF level in urine samples from 27 infants and young children with proliferative cutaneous hemangioma before and after treatment with low-dose superficial X-ray,and from 30 normal control infants and young children.Results The urine bFGF level was significantly higher in the patients before treatment than in those 3 months after the treatment and normal controls(t = 3.793,0.206,both P ＜ 0.05),while no significant difference was observed in the urine bFGF level between the treated patients and controls (t = 0.505,P ＞ 0.05).Conclusions Measurement of urine bFGF level by ELISA is a convenient and reliable method ,which may provide an objective index for the monitoring of hemangioma occurrence and development.     

Immunohistochemical analysis of keratinocyte growth factor and fibroblast growth factor 10 expression in psoriasis

The pathogenic mechanism underlying the hyperproliferation of keratinocytes in psoriasis is still not completely clarified. The production of cytokines released by activated T lymphocytes infiltrating the upper dermis probably has a crucial role. Even dermal fibroblasts can participate in the process through the secretion of growth factors, and some studies have reported an increased expression of the insulin-like growth factor 1. Few studies, however, have focused on the possible involvement of the keratinocyte growth factor (KGF/FGF-7) and the fibroblast growth factor 10 (FGF-10/KGF-2), which are secreted by fibroblasts and stimulate keratinocyte proliferation acting through a receptor specifically expressed by epithelial cells. The aim of this study was to investigate the expression of KGF and FGF-10 on the skin of patients with psoriasis by immunohistochemical analysis and to evaluate the correlation with the lymphocyte infiltrate and the epidermal proliferation. Immunostaining for KGF and FGF-10 showed that both the growth factors are upregulated in the upper dermis of psoriatic skin, and that the expression is correlated with the presence of T-cell infiltrate and with keratinocyte proliferation. Our data suggest that in psoriatic lesions activated lymphocytes can stimulate fibroblasts to produce KGF and FGF-10, which in turn contribute to sustain the hyperproliferative status of the keratinocytes.     

Reduced response of scleroderma fibroblasts to fibroblast growth factor

Summary Reactivity of scleroderma fibroblasts to lymphoid cell-derived fibroblast growth factor (FGF) was assessed in this study. The fibroblasts from the sclerotic lesion failed to respond to FGF, whereas those from scleroedematous lesions responded equally to normal fibroblast. Response of the fibroblast from sclerotic lesion was also lower than that of the fibroblast from mature scar. Fibroblasts obtained from three different layers of healthy skin, papillary dermis, reticular dermis, and reticular-subcutaneous layer, responded equally to FGF, whereas the fibroblast of reticular dermis from sclerotic lesion failed to respond to FGF. It is suggested that the fibroblast of reticuar dermis in scleroderma is variously activated by some unknown factors, so that they do not have enough reserve to respond to further stimuli.     

Basic fibroblast growth factor stimulates the proliferation of human dermal fibroblasts via the ERK1/2 and JNK pathways

Background Basic fibroblast growth factor (bFGF, FGF‐2) has been described as a multipotent cytokine that regulates cell growth as well as differentiation, matrix composition, chemotaxis, cell adhesion and migration in numerous cell types. It is known that bFGF stimulates proliferation of cultured fibroblasts. However, the detailed mechanism of fibroblast proliferation induced by bFGF in vitro still remains to be elucidated. Objectives We investigated the precise effects of bFGF on fibroblast proliferation and the signalling pathways responsible for bFGF‐induced proliferation in cultured human dermal fibroblasts (HDFs). Methods HDFs were cultured with bFGF in the presence or absence of specific inhibitors against MAPK signalling pathways including ERK, JNK and p38. The number of cells was counted and immunoblotting findings were examined for the activation of ERK1/2 and JNK. Furthermore, the inhibitory effects of ERK1, ERK2 and JNK1 were proven by the transfection of siRNA. Results bFGF increased the number of HDFs in a dose‐ and time‐dependent manner. The bFGF‐induced proliferation was suppressed by the MEK inhibitors PD98059 and U0126, and the JNK inhibitor SP600125. bFGF increased the phosphorylation levels of ERK1/2 and JNK1. Treatment with ERK1, ERK2 or JNK1 siRNA significantly inhibited bFGF‐induced proliferation. Conclusions This study indicates that ERK1/2 and JNK pathways play an important role in the bFGF‐mediated effect in HDFs. This study also suggests that controlling ERK1/2 and/or JNK signalling may therefore be a new therapeutic approach for the treatment of chronic and untreatable skin ulcers.     

皮肤瘙痒发病机制和药物治疗的新靶点

皮肤瘙痒的神经生理学机制一直未被阐明,传递瘙痒特定神经纤维亚单位的发现极大增加对瘙痒发病机制的认识,临床上抗组胺药物最常被用来治疗皮肤瘙痒,但对于改善患者慢性皮肤瘙痒的症状疗效不高,多种内源性物质包括生长冈子、神经肽、类花生酸类物质和细胞因子可以引起皮肤瘙痒.蛋白酶活化受体-2和辣椒素受体成为药物治疗的新靶点.     

Differential in vitro responses of elastin expression to basic fibroblast growth factor and transforming growth factor β1 in upper,middle and lower dermal fibroblasts

Elastin mRNA levels were measured in cultured skin fibroblasts derived from upper, middle and lower dermal layers. The elastin mRNA levels were highest in the fibroblasts from the upper dermal layer and lowest in the lower dermal fibroblasts. Modulation of elastin expression by basic fibroblast growth factor and transforming growth factor β1 in the dermal fibroblasts was also studied. Basic fibroblasts growth factor downregulated elastin expression in the upper dermal fibroblasts but did not significantly change elastin expression in the middle and lower dermal fibroblasts. Upregulation of elastin expression by transforming growth factor β1 was greater in the upper dermal fibroblasts than in the middle and lower dermal fibroblasts. Platelet-derived growth factor induced no significant changes in the three types of dermal fibroblasts. The results suggest that the differential responses of elastin expression to potent modulators may be at least partially responsible for the abnormal elastin metabolism specifically observed in the upper dermal layer. Received: 21 November 1995     

皮肤基底细胞癌组织中血管内皮细胞生长因子受体-2的表达

目的 研究血管内皮生长因子受体-2(VEGFR-2)在皮肤基底细胞癌(BCC)组织中的表达情况.方法 提取BCC组织的mRNA,以RT-PCR法检测标本中VEGFR-2mRNA的表达.同时应用蛋白质印迹法检测VEGFR-2蛋白的表达,免疫荧光法检测VEGFR-2在BCC组织中的定位.结果 BCC组织标本mRNA和蛋白水平中均可以检测到VEGFR-2的表达,并较正常人对照有显著增加.VEGFR-2在BCC细胞主要表现为膜性分布.结论 皮肤BCC组织细胞能够高异常表达VEGFR-2,可能是导致BCC细胞肿瘤性增殖的原因之一.     

Spatial and temporal expression of basic fibroblast growth factor protein during wound healing of rat skin

BACKGROUND: Basic fibroblast growth factor (bFGF) stimulates the mitogenesis of various cells and plays a key part in wound healing. OBJECTIVES: To determine the spatial and temporal expression of bFGF protein during wound healing after burning of rat skin. METHODS: Immunohistochemical methods were used. RESULTS: The immunostaining for bFGF in the normal epidermis was faint and sporadic in the basal cell layer. However, significant staining for bFGF was found in four locations: regenerated epidermis, a band-like zone near the regenerated epidermis, renewed capillaries, and cells infiltrating into the granulation tissue at the inflammatory to proliferative stages after the burn. The intensity of immunostaining of regenerated epidermis, the band-like zone and renewed capillaries was maximal during the proliferative stage and decreased to normal levels or disappeared simultaneously with wound closure. Immunopositive macrophage-like cell numbers in the granulation tissue increased during the proliferative stage and promptly decreased after wound closure, but such cells were only poorly visible in the scar tissue until 42 days postburn. CONCLUSIONS: bFGF may affect the proliferation, differentiation and migration of regenerated keratinocytes and the recruitment of inflammatory cells, as well as neovascularization in granulation tissue during wound healing. Macrophages may play a pivotal role in cutaneous wound repair by producing bFGF not only during the inflammatory or proliferative stages but also during the remodelling stage.     

Immunohistochemical localization of basic fibroblast growth factor in wound healing sites of mouse skin

Summary The immunohistochemical localization of basic fibroblast growth factor (bFGF) was examined during wound healing in mouse skin. Frozen sections taken from the rounded skin defects were reacted with polyclonal anti-human recombinant bFGF IgG followed by incubation with FITC-conjugated IgG. The basal layer keratinocytes and hair bulbs at the wound edge were strongly stained with this antibody. In the reepithelized area, several layers of keratinocytes from the basal layer were positively stained regardless of the time after wounding. These findings suggest that germinative keratinocytes which express bFGF function as leading cells in the covering of the wound defect. However, dermal granulation tissue, including capillary endothelial cells, fibroblasts and macrophages unexpectedly did not demonstrate any immunoreactivity throughout the process of wound healing. Simultaneous histochemical investigation using cultivated mouse keratinocytes and bovine aortic endothelial cells showed primarily cytoplasmic fluorescence. The discrepancy in the staining patterns of endothelial cells in vivo and in vitro suggests that immunoreactive bFGF is either not expressed in vivo, or is processed or masked.