• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of International Convention of the Pharmaceutical Society of Korea
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• pp.301.2-302
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• 2001

• Applied Microscopy
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• 제41권1호
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• pp.47-53
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• 2011

모발은 피부를 둘러싸고 있는 피부 부속기관이며, 외부로부터 들어오는 물리적, 화학적 자극에 대하여 보호하고, 미용 효과 등의 기능을 가지고 있다. 모발은 피부의 표피와 진피의 사이에서 발생하며, 모발성장주기를 가지고 있다. 모발성장주기는 모발이 성장하는 시기(anagen)와 성장을 끝내는 시기(catagen), 성장을 멈추고 휴식하는 시기(telogen)로 구분할 수 있다. 모발성장기 중, 성장기의 모낭 부위에는 많은 성장인자들이 분포하고 있다는 것이 이미 보고된 바 있다. 뇌하수체에서 분비되는 인간 성장 호르몬은 신진대사를 수행하는 신호를 전달하며, 기관의 성장을 촉진하는 역할을 수행한다. 성장호르몬은 혈류를 따라 온몸으로 이동되며, 여러 가지의 성장인자들을 활성하거나 억제하는 조절 작용을 한다는 연구결과가 보고된 바 있다. 본 연구에서는 인간성장호르몬을 유전자재조합기술을 이용하여 관찰이 용이하도록 6개의 히스티딘 아미노산을 첨가하였으며, 나노크기의 리포좀으로 캡슐화한 LhGH를 (주)리제론으로 부터 받아 사용하여 마우스(C57BL6/CrN)에서 LhGH에 의한 모발의 성장 관계를 알아보았다. 8주령의 마우스 3마리씩 증류수, Liposome, Minoxidil, His hGH, LhGH를 실험의 목적에 따라 20일, 6주 동안 도포하였고, 피부 속의 침투 경로는 형광면역반응법을 이용하여 형광현미경으로 관찰하였다. 또한, 광학현미경과 주사전자현미경을 이용하여 모발 수의 변화를 비교하여 보았다. 마우스 피부에 도포한 LhGH는 피부표피의 모공과 모발을 따라 피부 안으로 이동되는 것이 광학현미경으로 관찰되었다. 또한 LhGH는 성별에 관계없이 모발 수를 증가시켰으며, 피부 밖으로 도출된 모발 수를 증가시킴을 확인하였다. 자연적으로 탈모된 마우스에서도 발모를 증진시켰으며, 발모 후 3주 동안 새로운 탈모현상을 억제하였다. 이러한 결과로 LhGH는 마우스(C57BL/6CrN)에서 모발의 수를 증가시키며, 탈모된 피부에서 모발의 재성장을 촉진시키는 효과가 있다고 생각한다.

• BMB Reports
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• 제45권12호
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• pp.719-723
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• 2012

A close association between the obesity hormone leptin and breast cancer progression has been suggested. The present study investigated the molecular mechanism for enhanced leptin expression in breast cancer cells and its functional significance in breast cancer aggressiveness. We examined whether leptin expression level is affected by the oncoprotein human epidermal growth factor receptor2 (HER2), which is overexpressed in ~30% of breast tumors. Here, we report, for the first time, that HER2 induces transcriptional activation of leptin in MCF10A human breast epithelial cells. We also showed that p38 mitogen-activated protein kinase signaling was involved in leptin expression induced by HER2. We showed a crucial role of leptin in the invasiveness of HER2-MCF10A cells using an siRNA molecule targeting leptin. Taken together, the results indicate a molecular link between HER2 and leptin, providing supporting evidence that leptin represents a target for breast cancer therapy.

• 약학회지
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• 제57권2호
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• pp.125-131
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• 2013

Biologically active peptides, including growth factors and cytokines, participate in various biological processes in human skin. They could provide a great advantage of maintaining healthy skin. Many peptide growth factors like epidermal growth factor (EGF) and human growth hormone (hGH) have been used in cosmetic formulations. The delivery of peptide growth factors across the Stratum corneum, however, seems not sufficient because of their physical properties such as high molecular weight and hydrophilicity. So increasing the penetration of growth factors of interest into skin would be a major concern for ensuring their maximum biological efficacy. In this study, we have identified several skin penetration-enhancing peptides which facilitate delivery of growth factors, when fused at N-terminus of the target protein, into skin. For efficient and rapid screening, we constructed a skin-penetrating assay system using Franz cell and porcine skin. Next, we carried out phage display screening using M-13 bacteriophage with random 12 -amino acid library on its coat protein P3 on that system. After several selection rounds, peptide sequences facilitate the penetration of phages through the porcine skin were identified from a large population of phages. We found that phages with the most potent peptide (S3-2, NGSLNTHLAPIL) could penetrate the porcine skin eight times more than those with control peptide (12 mino acids scrambled peptide). Furthermore, growth factors conjugated with S3-2 peptide penetrate porcine skin three to five times efficiently than non-conjugated growth factors. In conclusion, our data shows that the skin penetration-enhancing peptide we have characterized could increase the delivery of growth factors and is useful for cosmeceutical application.

• 한국수정란이식학회지
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• 제13권2호
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• pp.97-106
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• 1998

The pronuclear injection of metallothionein-human growth hormone (MT-hGH) gene into rabbit zygotes was performed to establish in vitro developmental system and to detect the presence of the injected gene by nested PCR. Mature female New Zealand White rabbits were superovulated by eGG and hCG treatments. The rabbits were mated and the zygotes were collected from the oviducts 18-22 h after hCG injection by flushing with D-PBS. Two to three picoliters of MT-hGH gene was microinjected into male pronuclei. The foreign gene-injected zygotes were cultured in TCM-199 or RD mediurn containing 10% FCS with a monolayer of rabbit oviductal epithelial cefls in a 5% $CO_2$ incubator. The presence of injected DNA in rabbit embryos or blastomeres at different developmental stages .vas detected by a nested PCR analysis. The results are summarized as follows ; 1.The developmental rate of the MT-hGH gene-injected zygotes to blastocyst was significantly higher in TCM-199 medium (68.1%) than in RD medium (42.9%). 2.The gene injection into pronuclei at 18 or 22 hours post hCG treatment during pronuclear stage did not much affect on the in vitro development of the rabbit embryos. 3.The rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased when they developed to blastocysts. The results indicate that the screening of transgene in rabbit embryos by nested PCR analysis could be a prornisible method for the preselection of transgenic embryos. Furthermore, the preselection of transgenic embryos would greatly reduce hoth the cost and effort of production of transgenic animals.

• 약학회지
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• 제45권2호
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• pp.227-236
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• 2001

The complexity and variability of both the biologicals and the bioassays used to test them led to the use of the reference standard- a sample of the product of defined purity and potency, against which all preparations of that product must be calibrated. In order to prepare and establish KFDA reference standard for recombinant human growth hormone (somatropin), somatropin substance was filled in ampoules in National Institute for Biological Standards and Control (NIBSC). The candidate KFDA reference standard for somatropin (designated as 98/674) was evaluated to determine the suitability of serving as a KFDA reference standard for somatropin by the collaborative study, in which 10 laboratories participated. Physicochemical analysis and in vivo bioassay were performed by direct comparison with the international somatropin standard 88/624. 98/674 was identified as somatropin by SDS-PAGE, IEF, peptide mapping, and HPLC. Determination of somatropin content by SE-HPLC yielded a mean estimate of 2.01 mg somatropin per ampoule. Data from the study also yielded mean values of 0.39 $\pm$ 0.26% for high molecular weight impurities by SE-HPLC and mean values of 2.13 $\pm$ 1.29% for somatropin related proteins by RP-HPLC. Estimates of relative potency by weight gain bioassay in the hypophysectomised rats showed that relative potency of KS 98/674 was 1.07 aganist IS 88/624. Based on the results of the collaborative study, the candidate reference standard for somatropin is suitable to serve as a KFDA reference standard for somatropin.

• Clinical and Experimental Reproductive Medicine
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• 제48권3호
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• pp.245-254
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• 2021

Objective: In humans, polycystic ovary syndrome (PCOS) is an androgen-dependent ovarian disorder. Aberrant gene expression in folliculogenesis can arrest the transition of preantral to antral follicles, leading to PCOS. We explored the possible role of altered gene expression in preantral follicles of estradiol valerate (EV) induced polycystic ovaries (PCO) in a mouse model. Methods: Twenty female balb/c mice (8 weeks, 20.0±1.5 g) were grouped into control and PCO groups. PCO was induced by intramuscular EV injection. After 8 weeks, the animals were killed by cervical dislocation. Blood serum (for hormonal assessments using the enzyme-linked immunosorbent assay technique) was aspirated, and ovaries (the right ovary for histological examinations and the left for quantitative real-time polymerase) were dissected. Results: Compared to the control group, the PCO group showed significantly lower values for the mean body weight, number of preantral and antral follicles, serum levels of estradiol, luteinizing hormone, testosterone, and follicle-stimulating hormone, and gene expression of TGFB1, GDF9 and BMPR2 (p<0.05). Serum progesterone levels were significantly higher in the PCO animals than in the control group (p<0.05). No significant between-group differences (p>0.05) were found in BMP6 or BMP15 expression. Conclusion: In animals with EV-induced PCO, the preantral follicles did not develop into antral follicles. In this mouse model, the gene expression of TGFB1, GDF9, and BMPR2 was lower in preantral follicles, which is probably related to the pathologic conditions of PCO. Hypoandrogenism was also detected in this EV-induced murine PCO model.

• International Journal of Stem Cells
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• 제16권3호
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• pp.315-325
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• 2023

Background and Objectives: Glioblastoma (GBM) is an aggressive primary brain tumor characterized by its heterogeneity and high recurrence and lethality rates. Glioblastoma stem cells (GSCs) play a crucial role in therapy resistance and tumor recurrence. Therefore, targeting GSCs is a key objective in developing effective treatments for GBM. The role of Parathyroid hormone-related peptide (PTHrP) in GBM and its impact on GSCs remains unclear. This study aimed to investigate the effect of PTHrP on GSCs and its potential as a therapeutic target for GBM. Methods and Results: Using the Cancer Genome Atlas (TCGA) database, we found higher expression of PTHrP in GBM, which correlated inversely with survival. GSCs were established from three human GBM samples obtained after surgical resection. Exposure to recombinant human PTHrP protein (rPTHrP) at different concentrations significantly enhanced GSCs viability. Knockdown of PTHrP using target-specific siRNA (siPTHrP) inhibited tumorsphere formation and reduced the number of BrdU-positive cells. In an orthotopic xenograft mouse model, suppression of PTHrP expression led to significant inhibition of tumor growth. The addition of rPTHrP in the growth medium counteracted the antiproliferative effect of siPTHrP. Further investigation revealed that PTHrP increased cAMP concentration and activated the PKA signaling pathway. Treatment with forskolin, an adenylyl cyclase activator, nullified the antiproliferative effect of siPTHrP. Conclusions: Our findings demonstrate that PTHrP promotes the proliferation of patient-derived GSCs by activating the cAMP/PKA signaling pathway. These results uncover a novel role for PTHrP and suggest its potential as a therapeutic target for GBM treatment.

• 한국산학기술학회논문지
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• 제21권9호
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• pp.559-568
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• 2020

세포 분열 및 성장 촉진에 영향을 주는 상피세포 성장인자(Epidermal Growth Factor, EGF)는 다양한 의학적 용도를 갖고 있는 호르몬 단백질이다. 본 연구에서는 human EGF 유전자를 대장균 코돈에 최적화 하고 pRSET 벡터에 클로닝하여 발현벡터를 구축하였다. Human EGF를 봉입체가 아닌 활성이 있는 형태로의 과량 발현을 위해 코돈의 최적화와 더불어 최초로 샤페론 공발현이 시도되었다. 발현된 Native protein 형태의 재조합 human EGF는 고순도로 정제하기 위해 Ion Exchange Chromatography를 2번 연속적으로 수행하여 순수 분리 정제되었고, ELISA 분석결과 99% 이상으로 재조합 EGF의 활성도가 상업용 EGF와 유사하게 나타났으며, 세포증식시험 결과 인간 재조합 EGF는 인체 피부 섬유아세포의 세포증식을 촉진하는 것으로 확인 되었다. 본 연구의 인간 EGF 발현 시스템은 양적인 측면 뿐 아니라 성공적인 활성형태의 발현으로 추가적인 재접힘 과정 및 N 말단의 융합부분을 제거하기 위한 크로마토그래피 작업이 필요가 없다는 점에서 기존의 방법들에 대체 될 수 있는 효과적인 인간 EGF 발현 시스템을 제공하고 있다.

• Clinical and Experimental Pediatrics
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• 제52권11호
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• pp.1249-1259
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• 2009

목 적:소아 성장호르몬결핍증의 저신장 치료에 사용되는 성장호르몬 생물학적 제제인 서방형 주사제의 비용-효과성을 사회적 관점에서 기존 매일 주사제와 비교하여 평가하고자 한다. 방 법:성장 호르몬 치료를 받는 환아를 대상으로 시각화 척도 방법에 의해 건강 관련 효용을 측정하였다. 2008년 7월, 이 연구에 참여한 2개 병원에서 저성장증 치료를 받고 있는 149명의 환아 보호자에게 매일 주사제에 대한 효용과 주 1회 주사제를 사용할 경우 기대되는 효용을 인터넷을 통해 설문조사 하였다. 성장호르몬 주사요법 중 매일 주사제와 주 1회 주사제 투여에 소요되는 직․간접 비용은 차이가 없으므로, 두 제형의 의약품비만을 비교하였다. 결 과:매일 주사제에서 주 1회 주사제로 전환할 경우 예상되는 효용값은 0.584에서 0.784로 증가하며, 연평균 추가비용은 4,060,811원이 발생한다. 점증적 비용-효용비는 20,304,555원/QALY (quality-adjusted life year gained)로 계산되었다. 시나리오 분석 결과, 점증적 비용-효용비는 최소 15,751,198원/QALY에서 최대 25,489,929원/QALY이다. 결 론:Base case 및 시나리오 분석결과에서 제시하는 점증적 비용-효용비는 우리 나라 1인당 GDP의 0.85-1.37배 범위 내에 있으므로 매우 안정적으로 비용-효과성이 있는 것으로 판단된다. 따라서, 매일 주사제에서 주1회 주사제로의 투약방법 변경은 비용-효과적이라 할 수 있으며, 주 1회 주사제가 추가 비용을 부담하더라도 경제성이 우수하다고 결론지을 수 있다.