• Biomolecules & Therapeutics
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• 제3권1호
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• pp.80-84
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• 1995

Effects of titrated extract of Centella asiatica (TECA) and epidermal growth factor (EGF) isolated from the urine of pregnant horse on the proliferation of human epidermal keratinocyte in culture were studied. An increase in the number of keratinocyte was observed with the treatment of TECA at the concentration ranges from 1 $\mu\textrm{g}$/mι to 100 $\mu\textrm{g}$/mι. Effects of low molecular weight EGF (LEGF) and high molecular weight EGF (HEGF) on the proliferation of keratinocyte in culture were also studied. The number of keratinocyte in culture was significantly increased with LEGF and HEGF respectively at the concentration of 10 ng/mι. Simultaneous treatment of the keratinocyte with LEGF, HEGF and TECA led to the increased proliferation of keratinocytes resulting 96% of the effect of a positive control, EGF isolated from mouse submaxillary glands.

• 대한의생명과학회지
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• 제8권4호
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• pp.229-234
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• 2002

Chlorella is rich in chlorella growth factor (CGF). A review of the literature has described that CGF improves the capability of a Th1-based immunity, anticancer, antioxidant antibacterial activity, growth promotion, wound healing and so on, but has not studied the effect for the metabolism and the proliferation of human skin keratinocyte. The aim of this study was to examine the effect of metabolism and the proliferation of human skin keratinocyte in vitro. CGF was extracted with an autoclaving method which is a modified hot-water extraction method from dried chlorella and conformed by means of absorbance 0.22 at 260 nm. We have measured the extracellular acidification rate (ECAR) of the CGF by Cytosensor$^{\circledR}$ Microphysiometer and evaluated responsiveness depending upon the dosage on the HaCaT cell. The ECAR for the concentrations of 0.15, 1.5, 15, 150 $\mu\textrm{g}$/ml of CGF increased as a 103.6, 128.2, 149.0 and 423.9%, respectively compared to control (0.0 $\mu\textrm{g}$/ml, 100% ECAR). The ECAR for ErbBl tyrosine kinase inhibited by 4-anilinoquinazolines, $C_{16}$H$_{14}$BrN$_3$O$_2$.HCl on tile HaCaT cells with the amounts of 10 $\mu\textrm{g}$/ml of the CCF compared with 100 $\mu\textrm{g}$/ml of rhEGF. The conclusion of the study is that CGF might increase human epidermal keratinocyte proliferation through the interaction between the epidermal growth factor receptor and itself.

• 대한의생명과학회지
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• 제6권2호
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• pp.83-91
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• 2000

Protein kinase C는 세포의 신호전달계에 관여하는 중요한 조절효소로서 여러 가지 세포의 분화와 증식과도 밀접한 관련이 있다. 신생아의 포피 keratinocyte를 농도 200 ng/ml의 human recombinant epidermal growth factor (hrEGF)와 human recombinant insulin-like growth factor-1 (hrIGF-1) 그리고 hrEGF와 hrIGF-1의 혼합액을 각각 첨가하여 24시간 배양한후 세포질과 세포막의 PKC단백질을 추출하여 그 농도를 측정하고, Western blot analysis를 이 용하여 각 growth factor들의 PKC isoenzyme에 대한 영향을 분석하였다. 세포질의 총 PKC 단백질의 농도는 hrIGF-1을 처리한 keratinocyte에서 가장 높았으며, 세포막에서는 대조군의 단백질 농도가 가장 높게 나타났다. EGF를 처리한 keratinocyte의 세포질에서 는 PKC-$\beta$II, -$\delta$, -$\theta$가 막성분에서는 PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$, -$\theta$가 증가하였다. IGF-1을 처리한 군의 세포질성분에는 PKC-$\beta$I, -$\Im$, -$\theta$, 막성분에서는 PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$, -$\varepsilon$, -$\theta$가 증가하였다 EGF와 IGF-1의 혼합처리 군에서는, PKC-$\alpha$, -$\beta$I, -$\Im$, -$\theta$이 세포질에서, PKC-$\alpha$, -$\delta$, -$\Im$, -$\varepsilon$, -$\theta$은 세포막에서 증가하였다.

• 대한화장품학회지
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• 제49권1호
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• pp.75-85
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• 2023

본 연구에서는 7 개의 아미노산으로 이루어진 헵타펩타이드의 Lgr5 binding에 따른 인체 모낭 구성 세포의 활성에 대한 영향을 확인하였다. 표면 플라즈몬 공명(surface plasmon resonance, SPR) 시스템을 이용하여 헵타펩타이드가 Lgr5에 결합하는 것을 확인하였다. 인체 모유두세포(human hair follicle dermal papilla cell, HHFDPC)에 헵타펩타이드를 처리한 결과, 농도 의존적인 세포 증식이 나타났으며 β-catenin의 세포 내핵 이동 및 하위 유전자인 LEF1, Cyclin-D1, c-Myc의 발현 증가가 관찰되었다. 그리고 세포 증식 기전 관련 인자인 Akt와 ERK의 인산화 수준이 증가되었으며, 성장인자인 hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF) 발현이 유도되었다. 또한 인체 모모세포(human hair germinal matrix cell, HHGMC)의 분화 관련 전사 인자와 인체 외모근초세포(human hair outer root sheath cell, HHORSC)의 분화 표지 인자들도 헵타펩타이드 처리 시 높은 발현율을 보였다. 추가적으로 우리는 헵타펩타이드의 인체 모낭줄기세포(human hair follicle stem cell, HHFSC) 분화에 대한 영향을 조사하였다. 그 결과, HHFSC 표지인자들의 mRNA와 단백질 수준이 감소하였고 반면에 분화 표지인자들은 증가하였다. 상기의 결과들은 헵타펩타이드가 인체 모낭 구성 세포에서 Wnt/β-catenin 경로를 촉진시켜 증식 또는 분화를 유도할 수 있음을 보여준다. 이를 토대로 종합해 볼 때, 본 연구의 헵타펩타이드는 모발 성장을 유도하고 탈모 개선에 도움을 줄 수 있는 기능성 원료로 사용될 수 있을 것으로 보인다.

• 생명과학회지
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• 제21권6호
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• pp.907-911
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• 2011

인간형 재조합단백질 각질세포 성장인자를 안정적으로 생산하는 누에 배양 세포(Bm5-hKGF cell)을 만들었다. 이 세포에서 분비되어 배지에 포함된 양은 15-20 ng/ml 정도였다. Bm5-hKGF cell에 누에의 PDI를 함께 발현시키면 세포외 분비량이 2배 증가하였다. Wound healing migration assay 결과 누에세포에서 생산된 인간형 재조합단백질 각질세포 성장인자는 세포생장을 촉진하는 활성을 가지고 있었다. 본 실험의 결과는 누에배양세포를 사용하여 저비용으로 양질의 인간형 재조합단백질을 대량생산 할 수 있는 것을 기대한다.

• 한국산학기술학회논문지
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• 제14권4호
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• pp.1857-1862
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• 2013

교원질을 비롯한 세포외기질의 생합성을 통해 피부장력과 탄력 등 피부 특성을 제공하는 진피섬유모세포는 피부노화와 함께 활성이 감소되어 주름 형성의 이유가 된다. 따라서 젊고 건강한 피부를 유지하기 위해서는 진피섬유 모세포의 활성화가 큰 의미를 지닌다. 본 연구에서는 대복피 에탄올추출물이 진피섬유모세포의 세포외기질 합성에 미치는 영향을 ELISA, Western blot analysis 및 RT-PCR 등의 in vitro 평가법으로 측정하였다. ELISA와 western blot analysis에서 대복피추출물은 제1형 교원질, fibronectin, transforming growth factor-${\beta}1$ (TGF-${\beta}1$)의 생성을 촉진시켰고, RT-PCR에서는 COL1A1, TGF-${\beta}1$, keratinocyte growth factor (KGF), insulin growth factor (IGF)-1의 유전자 발현을 증가시켰다. 이상의 결과로부터 대복피추출물은 진피섬유모세포에서 세포외기질의 생성을 촉진시키는 천연소재인 것으로 판단되었다.

• KSBB Journal
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• 제16권3호
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• pp.237-240
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• 2001

본 연구에서는 keratinocytes 세포를 이용하여 VEGF가 세포의 증식, MMP-2 및 plasmin 분비에 미치는 영향을 조사하였다. 그 결과 VEGF는 keratinocytes의 세포증식을 약 2.5배 상승시켜 내피세포 뿐반 아니라 상피세포에서도 강력한 mitogen임을 확인하였으며 submaximal effect를 보이는 농도는 10 ng/mL이었다. VEGF의 농도 증가에 따른 MMP-2의 분비에 미치는 효과 역시 10 ng/mL에서 최대 효과를 나타냈으며 VEGF 처리 후 1시간만에 확인 가능한 MMP-2 밴드가 나타났다. 한편, MMP-2의 분비 증가와 함께 plasmin의 분비 증가가 수반되는지를 확인한 결과 VEGF를 첨가했을 때 약 3배 정도 plasmin의 분비량이 증가함을 알 수 었었다. 이러한 결과는 VEGF의 성장촉진 효과와 관련자어 볼 때 MMP-2와 plasmin이 keratinocytes 세포의 성장과 이동에 부분적으로 관여할 것임을 시사하고 있다.

• Toxicological Research
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• 제12권2호
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• pp.271-275
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• 1996

Carcinogenic potential of HPV-16 DNA and NNK in a human keratinocyte cell line was assessed to study effects of viral-chemical interaction. Human cells were transfected with HPV-16 DNA and 6 clonal cell lines were subsequently obtained. Clonal line-3 and 6 at passage 7 showed characteristics of tumor cells such as increases of saturation density, soft-agar colony formation, cell aggregation and foci appearance. Among cells treated with 1$\mu M$, 10$\mu M$, 100$\mu M$ or 1 mM of NNK for 4 weeks, 100$\mu M$ treatment showed most tumorigenic characteristics at passage 7. These results indicate that either HPV-16 or NNK alone is tumorigenic in this in human in vitro model. When cells transfected with HPV-16 were subsequently exposed by 100 uM NNK for 4 weeks, all the clonal cells except clone-1 showed higher levels of tumor cell characteristics than HPV-16 DNA or NNK exposure alone. Clonal line-6, the most tumorigenic cells, showed higher transcriptional level of fibronectin and lower level of TGF-$\beta_1$, as compared to control cells, suggesting that alteration of growth factor or extracellular matrix may play a role in carcinogenesis process induced by HPV-16 and NNK. Taken together, the present study indicates that viral-chemical interactions between HPV-16 DNA and NNK enhance carcinogenic potentials of human cells and implies that smoking among people infected with human papillomavirus may pose an additional risk of causing cancer.

• IMMUNE NETWORK
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• 제10권2호
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• pp.76-81
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• 2010

Ultraviolet B (UVB) induces multiple inflammatory and carcinogenic reactions. In skin, UVB induces to secrete several kinds of inflammatory cytokines from keratinocytes and also increases angiogenic process via the modulation of vascular endothelial growth factor (VEGF) production. Interleukin-21 (IL-21) is an inflammatory cytokine and produced by activated T cells. The biologic functions of IL-21 have not yet extensively studied. In the present study, we investigate the production of IL-21 from human keratinocyte cell line, HaCaT and its biological effect after exposure to UVB. First, we confirmed the IL-21 production and its receptor expression in HaCaT. And then, the change of IL-21 and VEGF production in HaCaT by UVB irradiation was examined. Not only IL-21 but also VEGF production was enhanced by UVB irradiation. Next, to determine relationship of enhanced production of IL-21 and VEGF, we detected VEGF production after neutralization of IL-21. VEGF production was reduced by IL-21 neutralization, which indicates that the IL-21 is involved in the VEGF production. Taken together, our results suggest that IL-21 and VEGF production is enhanced by UVB irradiation in HaCaT. In addition, it seems that IL-21 plays a role in the angiogenic process in skin via the modulation of VEGF production.

• The Korean Journal of Physiology and Pharmacology
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• 제6권5호
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• pp.275-279
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• 2002

Nerve growth factor (NGF) is an important autocrine growth factor and also a survival factor for keratinocytes. NGF may act in the hyperproliferative condition, psoriasis. Clinically, the combination of psoralen and UVA (PUVA) has been used in the treatment of a wide variety of cutaneous disorders, such as psoriasis and vitiligo. However, the precise therapeutic mechanism of PUVA on the dermatologic diseases remains unclear. The purpose of this study was to examine whether the expression of NGF in cultured keratinocytes is influenced by PUVA. Thus, normal human keratinocytes were isolated from neonatal foreskin, and the third to fifth-passaged cells were used in this study. The cells were exposed to various doses of UVA (30, 60, 120 $mJ/cm^2)$ after adding 8-methoxypsoralen (8-MOP) to examine the expression of NGF mRNA. The RNA and protein of the cells were extracted at various time points (1, 8, 24 hours) after UVA irradiation to examine the expression of NGF mRNA and production of NGF protein. In keratinocytes, there were no differences in the expression of NGF mRNA between the different doses of UVA irradiation, however, the expression of NGF mRNA in UVA and PUVA groups tended to increase as the time increased. The expression of NGF mRNA was the highest in PUVA group, followed by UVA group and the lowest in 8-MOP group. The expressions of NGF protein at 1 and 8 hours after UVA irradiation were lower in the PUVA group than in the other groups. This study showed that the expression level of NGF protein in keratinocytes was relatively lower in the PUVA groups than in the other groups, suggesting that the therapeutic mechanism of PUVA in psoriasis is related to the decrease of NGF protein.