Cardiac Ankyrin Repeat Protein의 과량발현이 혈관내피세포에서 갖는 혈관신생 촉진 효과

공훈영,변종회,Kong, Hoon-Young,Byun, Jong-Hoe 한국미생물학회 2008 미생물학회지 Vol.44 No.4

혈관이 좁아지거나 막혀서 생기는 허혈은 심장, 뇌, 다리와 같은 인체의 여러 장기에 영향을 미친다. 최근 혈관신생 분야에서 많은 진전이 있어 기존 치료법으로 치료가 되지 않는 허혈성 환자들의 치료 가능성에 대한 기대가 많아졌다. 혈관형성은 여러 개의 인자들과 세포들이 관여하는 복잡한 과정이기 때문에, 한 개의 인자보다는 여러 인자들을 병합하는 요법이 점점 많이 시도되고 있는데, 이런 병합요법의 한 예로 전사인자를 전달하는 전략을 생각할 수 있다. 이에 본 연구에서는 cardiac ankyrin repeat protein (CARP)의 유전자를 대상으로 그 혈관신생 능력을 혈관내피세포에서 조사하였다. 아데노바이러스 벡터 내에 human CARP의 cDNA를 클로닝하여 재조합 아데노바이러스를 제조하였으며, 이를 이용한 유전자 전달실험 결과, CARP 유전자 전달 군에서 유의하게 혈관내피세포의 중식과 모세관 구조 형성, 그리고 vascular endothelial growth factor의 발현 등을 증가시킴을 확인하였다. 본 연구 결과는 CARP가 혈관신생 연구의 새로운 목표 유전자로서 그 기능에 대한 많은 연구가 필요함을 뒷받침해준다. Tissue ischemia resulting from the constriction or obstruction of blood vessels leads to an illness that may affect many organs including the heart, brain, and legs. In recent years, considerable progress has been made in the field of therapeutic angiogenesis and the new approaches are expected to cure those "no-option patients" who are unsuited to conventional therapies. Although single angiogenic growth factor may be successful in inducing angiogenesis, combination of multiple growth factors is increasingly sought these days to augment the therapeutic responses. This trend is proper in light of the fact that blood vessel formation is a complex and multi-step process that requires the actions of many different factors. To meet the growing need for functionally significant blood flow recovery in the ischemic tissues, a novel strategy that can provide concerted actions of multiple factors is required. One way to achieve such a goal is to use a transcription factor that can orchestrate the expression of multiple target genes in the ischemic region and thus induce significant level of angiogenesis. Here, a putative transcription factor, cardiac ankyrin repeat protein (CARP), was evaluated in adenoviral vector context for angiogenic activity in human umbilical vein endothelial cells. The results indicated significant increase in proliferation, capillary-like structure formation, and induction of vascular endothelial growth factor, a typical angiogenic gene. Taken together, these results suggest that CARP represents itself as a novel target for therapeutic angiogenesis and warrants further investigation.

Differential Effects of Cysteine and Histidine-Capped ZnS:Mn Nanocrystals on Escherichia coli and Human Cells

공훈영,Song-Yi Kim,변종회,황청수 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.1

Cysteine and histidine-capped water-dispersible ZnS:Mn nanocrystals (ZnS:Mn-Cys and ZnS:Mn-His) were synthesized and their effects on E. coli and human cells were investigated. Particle sizes of these nanocrystals were found from HR-TEM images to be 3.5 nm and 4.0 nm, respectively. Their solution photoluminescence spectra showed identical broad emission peaks at 580 nm. ZnS:Mn-His significantly suppressed the growth of E. coli at 100 μg/mL and 1 mg/mL concentrations, something not observed with ZnS:Mn-Cys. Consistent with this, greater inhibition of cell proliferation and viability were observed in HEK293 and IMR90 cells in ZnS:Mn-His at 100 μg/mL and 1 mg/mL concentrations.

Syntheses of Biologically Non-Toxic ZnS:Mn Nanocrystals by Surface Capping with O-(2-aminoethyl)polyethylene Glycol and O-(2-carboxyethyl)polyethylene Glycol Molecules

공훈영,Byungkwan Song,변종회,황청수 대한화학회 2013 Bulletin of the Korean Chemical Society Vol.34 No.4

Water-dispersible ZnS:Mn nanocrystals were synthesized by capping the surface of the nanocrystal with O-(2- Aminoethyl)polyethylene glycol (PEG-NH2, Mw = 10,000 g/mol) and O-(2-Carboxyethyl)polyethylene glycol (PEG-COOH, Mw = 10,000 g/mol) molecules. The modified PEG capped ZnS:Mn nanocrystal powders were thoroughly characterized by XRD, HR-TEM, EDXS, ICP-AES and FT-IR spectroscopy. The optical properties were also measured by UV/Vis and photoluminescence (PL) spectroscopies. The PL spectra showed broad emission peaks at 600 nm with similar PL efficiencies of 7.68% (ZnS:Mn-PEG-NH2) and 9.18% (ZnS:Mn- PEG-COOH) respectively. The measured average particle sizes for the modified PEG capped ZnS:Mn nanocrystals by HR-TEM images were 5.6 nm (ZnS:Mn-PEG-NH2) and 6.4 nm (ZnS:Mn-PEG-COOH), which were also supported by Debye-Scherrer calculations. In addition, biological toxicity effects of the nanocrystals over the growth of wild type E. coli were investigated. They showed no biological toxicity to E. coli until very high concentration dosage of 1 mg/mL of the both nanocrystal samples.

Biological Toxicity Changes of Mercaptoacetic Acid and Mercaptopropionic Acid Upon Coordination onto ZnS:Mn Nanocrystal

공훈영,황청수,변종회 대한화학회 2012 Bulletin of the Korean Chemical Society Vol.33 No.2

Mercaptoacetic acid (MAA) and mercaptopropionic acid (MPA) capped ZnS:Mn nanocrystals were synthesized and their physical characteristics were examined by XRD, HR-TEM, EDXS, and FT-IR spectroscopy. The optical properties of the MPA capped ZnS:Mn nanocrystals dispersed in aqueous solution were also measured by UV/Vis and solution photoluminescence (PL) spectra, which showed a broad emission peak around 598 nm (orange light emissions) with calculated relative PL efficiency of 5.2%. Comparative toxicity evaluation of the uncoordinated ligands, MAA and MPA, with the corresponding ZnS:Mn nanocrystals revealed that the original ligands significantly suppressed the growth of wild type E. coli whereas the ligandcapped nanocrystals did not show significant toxic effects. The reduced cytotoxicity of the conjugated ZnS:Mn nanocrystals was also observed in NIH/3T3 mouse embryonic fibroblasts. These results imply that potential toxicities of the capping ligands can be neutralized on ZnS:Mn surface.

Prostatic acid phosphatase의 전립선 암에서의 역할

Hoon Young Kong(공훈영),Hak Jong Lee(이학종),Jonghoe Byun(변종회) 한국생명과학회 2011 생명과학회지 Vol.21 No.6

Prostatic acid phosphatase (PAP)는 전립선 암의 진단에 널리 사용되는 표지자로서 1935년 처음으로 동정되었고 인체 전립선에 가장 많이 존재하는 탈 인산화효소이다. PAP는 prostate epithelial cells에서 합성되는 전립선 특이적인 효소로서, 산성 환경에서 효소활성을 띠는 acid phosphatase 그룹에 속한다. PAP는 전립선액에 풍부히 존재하여 수정, 정자부족증, 만성통증의 감소에 관여한다. 그러나 가장 눈에 띄는 기능은 ERK1/2와 MAPK 경로에 관계된 HER-2와 PI3P의 탈 인산화를 유도하여 세포 성장 신호를 억제하고 전립선 암의 억제자로 작용하는 것이다. 최근 PAP DNA 백신을 이용하는 임상시험이 현재 진행 중이고, PAP를 이용한 immunotherapy를 통해 전립선 암을 치료하는 방법이 FDA의 승인을 받아 시행되고 있다. 이러한 PAP의 임상적 중요성에도 불구하고 현재까지 PAP의 분자적 조절기작에 대한 이해는 제한적이라 PAP에 대한 많은 연구가 필요한 실정이다. PAP는 NF-κB, TNF-α, IL-1 및 androgen과 androgen receptor에 의하여 promoter region이 조절된다고 알려졌다. 본 총설에서는 현재까지 밝혀진 PAP 유전자 및 단백질의 특징들과 더불어 전립선 암에서의 PAP의 기능, 발현 조절, 역할들을 종합하였다. Prostatic acid phosphatase (PAP) is one of the widely used biomarkers in the diagnosis of prostate cancer. It was initially identified in 1935 and is the most abundant phosphatase in the human prostate. PAP is a prostate-specific enzyme that is synthesized in prostate epithelial cells. It belongs to the acid phosphatase group that shows enzymatic activity in acidic conditions. PAP is abundant in prostatic fluid and is thought to have a role in fertilization and oligospermia. It also has a potential role in reducing chronic pain. But one of the most apparent functions of PAP is the dephosphorylation of macromolecules such as HER-2 and PI3P that are involved in the ERK1/2 and MAPK pathways, which in turn leads to inhibition of cell growth and tumorigenesis. Currently, clinical trials using PAP DNA vaccine are underway and FDA-approved immunotherapy using PAP is commercially available. Despite these clinically important aspects, molecular mechanisms underlying PAP regulation are not fully understood. The promoter region of PAP was reported to be regulated by NF-κB, TNF-α, IL-1, androgen and androgen receptors. Here, the features of PAP gene and protein structures together with the function, regulation and roles of PAP in prostate cancer are discussed.

Ultrasound contrast-enhanced study as an imaging biomarker for anti-cancer drug treatment: preliminary study with paclitaxel in a xenograft mouse tumor model (secondary publication)

이학종,황성일,변종회,공훈영,정현숙,강미라 대한초음파의학회 2017 ULTRASONOGRAPHY Vol.36 No.4

Purpose: The purpose of this study was to assess tumor angiogenesis using contrast-enhanced ultrasonography (CEUS) of human prostate cancer cells (PC3) that were implanted in mice before and after paclitaxel injection. Methods: Twelve mice were injected with human PC3. The mice were grouped into two groups; one was the paclitaxel-treated group (n=6) and the other was the control group (n=6). Before administering paclitaxel into the peritoneal cavity, baseline CEUS was performed after the administration of 500 μL (1×108 microbubbles) of contrast agent. The area under the curve (AUC) up to 50 seconds after injection was derived from the time-intensity curves. After injection of paclitaxel or saline, CEUS studies were performed at the 1-week follow-up. Changes in tumor volume and the AUC in both two groups were evaluated. After CEUS, the microvessel density (MVD) was compared between the groups. Results: In the paclitaxel-treated group, the AUC from CEUS showed a significant decrease 1-week after paclitaxel administration (P=0.030), even though the tumor volume showed no significant changes (P=0.116). In the control group, there was no significant decrease of the AUC (P=0.173). Pathologically, there was a significant difference in MVD between both groups (P=0.002). Conclusion: The AUC from the time intensity curve derived from CEUS showed an early change in response to the anti-cancer drug treatment that preceded the change in tumor size. The findings of CEUS could serve as an imaging biomarker for assessing tumor responses to anti-cancer drug treatment.

항암제 치료에 있어서 영상 생체표지자로서의 초음파 조영제 영상검사: 파크리탁셀과 이종이식 종양 모델을 이용한 예비연구

이학종,황성일,변종회,공훈영,정현숙,강미라 대한초음파의학회 2011 ULTRASONOGRAPHY Vol.30 No.2

Purpose: We wanted to assess tumor angiogenesis of human prostate cancer cells (PC3) implanted in mice before and after paclitaxel injection via contrast-enhanced ultrasonography (CEUS). Materials and Methods: Twelve mice were injected with human prostate cancer cells (PC3) on the back or hind limbs. The mice were grouped into two groups; one was the paclitaxel treated group (n = 6) and the other was the control group, which was treated with normal saline (n = 6). Before injection of paclitaxel into the peritoneal cavity, baseline CEUS was performed by the administration of 500 μl (1×108microbubbles) of contrast agent. The area under the curve (AUC) up to 50 seconds after contrast injection was derived from the time-intensity curves. After injection of paclitaxel or saline, one week follow up CEUS studies were performed. The changes of the tumor volume and the AUC in both two groups were evaluated. After CEUS, the mice were sacrificed and the microvessel density (MVD) was compared. Results: In the paclitaxel treated group, the AUC from CEUS showed a significant decrease one week after paclitaxel administration (p = 0.03), even though the tumor volume showed no significant changes (p = 0.116). In the control group, there was no significant decrease of the AUC (p = 0.173). Pathologically, there was a significant difference of microvessel density in both groups (p = 0.002). Conclusion: The AUC from the time intensity curve derived from CEUS showed early change in response to the anti-cancer drug treatment in advance of a tumor size response. The findings of CEUS could be an imaging biomarker for assessing the tumor response to anti-cancer drug treatment.

Identificaiton of Novel Immunogenic Human Papillomavirus Type 16 E7-Specific Epitopes Restricted to HLA-A*33;03 for Cervical Cancer Immunotherapy

김성훈,임종백,정혜원,공훈영 연세대학교의과대학 2017 Yonsei medical journal Vol.58 No.1

Purpose: To identify new immunogenic HLA-A*33;03-restricted epitopes from the human papillomavirus (HPV) 16 E7 proteinfor immunotherapy against cervical cancer. Materials and Methods: We synthesized fourteen overlapping 15-amino acid peptides and measured intracellular interferon-γ(IFN-γ) production in PBMC and CD8+ cytotoxic T lymphocytes (CTLs) after sensitization with these peptides using flow cytometryand ELISpot assay. The immunogenicity of epitopes was verified using a 51Cr release assay with SNU1299 cells. Results: Among the fourteen 15-amino acid peptides, E749-63 (RAHYNIVTFCCKCDS) demonstrated the highest IFN-γ productionfrom peripheral blood mononuclear cells (PBMCs), and CD8+ CTLs sensitized with E749-63 showed higher cytotoxic effect againstSNU1299 cells than did CD8+ CTLs sensitized with other peptides or a negative control group. Thirteen 9- or 10-amino acid overlappingpeptides spanning E749-63, E750-59 (AHYNIVTFCC), and E752-61 (YNIVTFCCKC) induced significantly higher IFN-γ productionand cytotoxic effects against SNU1299 cells than the other peptides and negative controls, and the cytotoxicity of E750-59- and E752-61-sensitized PBMCs was induced via the cytolytic effect of CD8+ CTLs. Conclusion: We identified E750-59 and E752-61 as novel HPV 16 E7 epitopes for HLA-A*33;03. CD8+ CTL sensitized with these peptidesresult in an antitumor effect against cervical cancer cells. These epitopes could be useful for immune monitoring and immunotherapyfor cervical cancer and HPV 16-related diseases including anal cancer and oropharyngeal cancer.