Synthetic Bile Acid Derivative HS-1200-induced Apoptosis of Human Osteosarcoma Cells

김규천(Gyoo-Cheon Kim),허용수(Young-Soo Her),박재현(Jae-Hyun Park),문용석(Yong-Suk Moon),유영현(Young-Hyun Yoo),신상훈(Sang-Hun Shin),박봉수(Bong-Soo Park) 대한해부학회 2004 Anatomy & Cell Biology Vol.37 No.5

담즙산 및 합성유도체가 여러 암세포에서 세포자멸사를 유도하여 항암활성을 보인다고 밝혀졌지만 뼈육종세포에서의 세 포자멸사 유도활성은 보고된 바 없다. 본 연구는 담즙산 유도체들이 뼈육종세포에 세포자멸사를 유도하는지를 알아보고 그 기작을 연구하기 위하여 수행되었다. UDCA 합성 유도체와는 달리 CDCA 합성유도체는 뼈육종세포의 사망을 유도하였다. CDCA 유도체 중 HS-1200을 선택하여 세포자멸사 기전을 연구하였다. HS-1200은 핵의 농축, cytochrome c 방출, Bax/Bcl-xL분율 변화, caspase-3의 활성화 및 CAD와 PARP 분절 등을 관찰되었다. 비록 더 많은 연구가 필요하지만 본 시험관 연구 자료는 합성담즙산 유도체 투여가 뼈육종의 치료전략이 될 수 있음을 시사한다. Bile acids and synthetic its derivatives induced apoptosis in various kinds of cancer cells and had anticancer effects. However, it wasn’t discovered those materials have apoptosis induced effects on osteosarcoma cells. The present study was done to examine the synthetic bile acid derivatives induced apoptosis on osteosarcoma cells and such these apoptosis events. The synthetic bile acid derivatives, chenodeoxycholic acid (CDCA) induced the cell death on human osteosarcoma (HOS) cells contrary to ursodeoxycholic acid (UDCA). HS-1200, a synthetic derivative of CDCAs, was chosen to experiment apoptosis events in HOS cells. HOS cells treated with HS-1200 showed nucleus condensation, cytochrom c release, Bax/Bcl-xL alteration, activation of caspase-3 and caspase-activated deoxyribonuclease (CAD), and degradation of poly (ADP-ribose) polymerase (PARP). Though this study needs more investigations, these in vitro data suggest that treatment of the synthetic bile acid derivatives can give medical therapy on HOS cells.

Hyaluronic Acid에 대한 사람 뼈육종의 부착에 있어서 CD44의 역할

김규찬(Gyoo-Cheon Kim),황윤(Yoon Hwang),박혜련(Hae-Ryoun Park),조기진(Gi Jin Jo),정준영(Jun-Young Chung),유영현(Young-Hyun Yoo),박봉수(Bong-Soo Park) 대한체질인류학회 2001 해부·생물인류학 (Anat Biol Anthropol) Vol.14 No.4

뼈모세포는 분화과정 중에 몇몇 세포밖바탕질 분자들을 단계적으로 발현하며, 이러한 사실은 다양한 부착기작들이 뼈모세포의 분화와 뼈발생을 조정하고 있음을 암시한다. 세포밖바탕질 중의 하나인 hyaluronicacid (HA) 분자는 연골와 세포밖바탕질에 주로 분표하는 glycosaminoglycan (GAG) 후서 매우 큰 분자량을 가지고 있으며 용매와 결합하여 높은 정성을 나타내게 된다. 발생초기와 조직 분화 전의 몇몇 조직에서 세포밖바탕질의 주된 구조적 거대분자를 구생하며, 세포의 증식과 이동을 가능하게 한다. CD44는 HA 에 대한 주된 수용기로 알려져 있으며, 다양한 세포의 표면에서 필수 막 당단백질의 형태로 발현 된다. 세포뼈대인 가는 미세섬유 (actin filament) 와 세포밖바탕질사아에서 막통과신호전달 (transmembrane signalling) 의 가능이 있는 것으로도 알려져 있다. 본 실험에서는 뼈육종세포를 모델로 사용하여 이세포가 HA 에 대한 부착여부와 부착할 때 세포막에 존재하는 CD44 와 GAGs 의 관여여부, 그때 CD44 발현양상에 변화가 있는지를 알아보기 위하여 본 실험을 시행하였다. Has 는 HA 에 대하여 농도 의존적으로 부착하였으며, 부착한 후 180 분 경과 시에 세포의 약 90% 가 HA 에 부착하였다. CD44 와 hyaluronidase, genistein으로 Has 에 전처리 하였을 때 세포의 부착율은 현저하게 감소하였다. 그리고 HA 를 도포한 군에서 Has 는 HA 흉 도포하지 않은 군에 비해 훨씬 많은 CD44를 발현하였다. 이상의 결과를 고려하여 볼 때 Has 는 HA 에 대한 부착 친화성을 가지고 있었으며 이러한 부착에 있어서 세포막 단백질인 CD44 와 세포막에 존재하는 GAGs, 세포질에 존재하며 이차적으로 세포 신호전달계통에 참여하는 단백질인 tyrosine kinases 가 중요한 역할을 한다는 것을 설명하였다.

구강 편평세포 암종에서의 CD44 발현

박상준(Sang Jun Park),박혜련(Hae Ryoun Park),김규천(Gyoo Cheon Kim),박봉수(Bong Soo Park),김태규(Tae Kyu Kim) 대한구강악안면외과학회 2000 대한구강악안면외과학회지 Vol.26 No.2

The cell surface glycoprotein CD44 is a kind of adhesion molecule, which binds hyaluronic acid, type I collagen and fibronectin. Although there have been numerous reports on the expression and the function of CD44 in lymphocytes and macrophages, very little is known about its distribution and definite role in epithelial tissue, especially in oral epithelial one. The present study was performed to investigate the distribution and expression of the CD44 in human gingiva and squamous cell carcinoma(SCC) arising in human gingiva. And the authors compared CD44 expression with histopathologic grade of SCC. The results were as follows: 1. The CD44 was strongly expressed in granular, spinous and basal layers of normal marginal and attached gingiva, in spinous and basal layers of normal sulcular gingiva, and in all epithelial layers of normal junctional gingiva. 2. In SCC of gingiva, the CD44 was expressed in all but one case. In most of the cases the CD44 was expressed at cell membrane and the degree of expression was relatively strong. 3. In low-grade SCC of gingiva, the CD44 was strongly expressed, especially at the basal and spinous layers of abundantly keratinized cancer nests. In high-grade SCC of gingiva, the CD44 expression tended to be weak but was strong at cells showing individual keratinization. This study suggest that the CD44 expression of normal and cancerous gingival epithelium is associated with the degree of proliferation and differentiation of epithelial cells.

Apoptotic Response of Human Oral Squamous Carcinoma Cells to Etoposide

김규천,이경덕,박재현,김덕한,박정길,박준상,박봉수,Kim, Gyoo-Cheon,Lee, Kyoung-Duk,Park, Jae-Hyun,Kim, Duk-Han,Park, Jeong-Kil,Park, June-Sang,Park, Bong-Soo Korean Academy of Orofacial Pain and Oral Medicine 2005 Journal of Oral Medicine and Pain Vol.30 No.2

항암제의 연구는 화학물질에 민감한 암세포를 죽음에 이르게 하는 세포자멸사와 같은 다양한 세포기능에 초점을 맞추어 왔다. 그러나 약물이 유도한 세포의 죽음에 있어서 핵심적인 분자적 기작은 아직 잘 이해되지 않고 있다. Etoposide는 폐암과 고환암에 사용되는 항암제로서, 본 연구는 etoposide가 사람구강편평상피암종세포(OSC9)에도 세포독성효과와 세포자멸사를 일으키는지를 알아보기 위해 실행하였다. 이 실험에서 etoposide는 농도와 시간 의존적으로 OSC9 세포의 생존율를 현저하게 저해시켰다. TUNEL 염색과 Hoechst 염색을 이용한 핵의 형태학적 관찰에서는 etoposide에 의해 핵이 응축되고 분절되었다. p53의 발현은 48 시간에 증가했으며, etoposide 처리로 인해 caspase-3의 활성을 관찰할 수 있었으며, 그 기질에 해당되는 PARP 단백질은 116-kDa과 89-kDa으로 분절되었다. 위의 결과들은 OSC9 세포에서 etoposide가 유도한 세포자멸사는 caspase-3의 활성과 관련됨을 설명하고 있다.

Bolting Resistant and High-Yielding Angelica acutiloba Cultivar, "Jinil"

Dong Hwi Kim,Hong Seob Yu,Hee Woon Park,Chun Geun Park,Jung Sook Sung,Chung Heon Park,Nak Sul Seong,Geum Soog Kim,Oh Heun Kwon,Cheon Gyoo Park,Young Min Jin 한국육종학회 2006 한국육종학회지 Vol.38 No.3

Angelica acutiloba cultivar developed by a medicinal crop breeding teamof the National Institute of Crop Science during the period from 1993 to 204. The cultivar was selected from Jinbu local variety.Jinil has a green stem color. It has a broader leaf bla

Effects on Skin Irritation and Turnover Rate by the Control of Skin Permeability of Alpha-hydroxyacids

( Cheon-koo Lee ),( Seong-don Hong ),( Mun-eok Park ),( Sun-gyoo Park ),( Seong-hoon Jeong ),( Seh-hoon Kang ) 대한화장품학회 1996 대한화장품학회지 Vol.22 No.2

The effect of a novel delivery system, water in oil emulsion containing chitosan hydrogel as a inner phase (W/O-C) was evaluated, and the relationships between the skin permeation, the skin primary irritation and the skin turnover rate of AHAs were discussed. We selected glycolic acid (GA), lactic acid (LA), malic acid (MA), and tartaric acid (TA) as model AHAs. The steady state fluxes of 4 AHAs across the excised hairless mouse skin increased as the molecular weights of the AHAs decreased (GA>LA>MA>TA). The skin turnover times were shortened in all AHAs, compared with control. The skin permeation and the skin primary irritation of the LA decreased and the skin turnover time increased, as the pH increased. The maximum therapeutic index was obtained with pH 3.8, 0.5 M LA. It was suggested that the skin permeability of LA might be a main factor for prediction of the skin irritation and the skin turnover time. On the other hand, the W/O-C containing pH 3.8, 0.5 M LA indicated a good sustained release property of LA, compared with water in oil emulsion without chitosan hydrogel (W/O) or oil in water emulsion (O/W). The skin permeability and the skin irritation of AHAs from the W/O-C decreased, compared with W/O or O/W, however the skin turnover time showed almost the same value as W/O or O/W. In conclusion, we suggest that the control of the skin permeation of AHAs would be an important tool for reducing the skin irritation and for maintaining the positive effect of AHAs, and the W/O-C system could be a potential candidate for future cosmetological application of AHAs.

Production of Hexanoic acid from Kluyveromyces marxianus

Yuna CHEON,Jun-Seob KIM,Jun-Bum PARK,JaeHyung LIM,Gyoo Yeol JUNG,Jin-Ho SEO,Jin-Byung PARK,Suk-Jin HA,Dae-Hyuk KWEON 한국생물공학회 2013 한국생물공학회 학술대회 Vol.2013 No.10

Induction of Apoptosis in Human Oral Squamous Carcinoma Cells by Extracellular Products from Pseudomonas aeruginosa

김규천(Gyoo-Cheon Kim),강현희(Hyeun-Hee Kang),김현철(Hyeon-Cheol Kim),김인령(In-Ryeon Kim),이무형(Moo-Hyung Lee),구병찬(Byung-Chan Koo),김덕한(Duk-Han Kim),민지학(Ji-Hak Min),박봉수(Bong-Soo Park) 대한해부학회 2005 Anatomy & Cell Biology Vol.38 No.4

100년 전부터 암을 가진 동물이나 사람이 박테리아에 감염되었을 때, 그 암조직이 줄어들었다는 보고가 있었다. 박테리아는 감염된 세포내에서 단백질합성 저해제와 구멍형성 단백질 분비, 죽음에 이르게 하는 내재성 단백질 분자의 활성화를 포함하는 다양한 기작을 통해서 세포자멸사를 유발할 수 있다. 본 연구는 Pseudomonas aeruginosa 균주로부터 분리한 세포밖 분비물(EPPA)이 구강편평상피암종세포(OSC9)의 세포자멸사를 일으키는지를 알아보고자 시행 하였다 EPPA는 OSC9 암세포에 농도 의존적으로, 또한 시간 의존적으로 암세포독성효과를 나타내었다. Hoechst 염색법, TUNEL 분석법 그리고 DNA 전기영동 등의 방법을 통해 염색질 응축과 핵분절 등이 확인되었다. EPPA가 처리 후 24시간 경과된 세포에서부터 caspase-3와 caspase-6의 활성형태를 관찰할 수 있었다. Caspase-3의 기질인 PARP와 DFF45 그리고 caspase-6의 기질인 lamin A의 분절형태가 관찰 되었다. 이상의 결과로 EPPA가 OSC9 암세포에서 casapse 의존성 세포자멸사를 유발시킴을 알 수 있었다. 사람구강편 평상피암종에 항암효과를 가지는 새로운 물질이 EPPA에 포함되어 있는 것으로 보인다. It was reported that cancer in humans and animals infected with microbial pathogens was regressed about 100 years ago. Bacteria are able to trigger apoptosis by a variety of mechanisms including the secretion of protein synthesis inhibitors, pore forming proteins, molecules activating the endogenous death machinery in the infected cell. This study was conducted in order to investigate whether extracellular products of Psuedomonas aeruginosa (EPPA) induce apoptosis in human oral carcinoma cells (OSC9). The EPPA showed cytotoxic effect on OSC9 cells in dose and time-dependent manner. The cell death was demonstrated to be due to apoptosis characterized by chromatin condensation and nuclear fragment. EPPA treatment induced cleavage of caspase-3 and caspase-6. The caspase substrates, PARP, DFF45 and lamin A were cleaved during EPPA-induced apoptosis. Taken together, EPPA induces apoptosis on human oral squamous carcinoma cells in caspase-dependent manner. Our data therefore provide that EPPA contains a novel antitumor agent for human oral squamous carcinoma.

Apoptotic Effect of Combination Treatment with a Proteasome Inhibitor, Lactacystin, and a Histone Deacetylase Inhibitor, Trichostatin A, on MCF-7 Cells

김규천(Gyoo-Cheon Kim),신유정(Yoo-Jung Shin),김인령(In-Ryeon Kim),이무형(Moo-Hyung Lee),황영섭(Young-Seob Hwang),길영기(Young-Gi Gil),박봉수(Bong-Soo Park) 대한해부학회 2006 Anatomy & Cell Biology Vol.39 No.6

Proteasome 저해제와 HDAC 저해제의 상호작용에 의한 상승효과가 여러 형태의 암에서 세포자멸사를 일으키는 연구가 많음에도 불구하고, 그 기작에 대해서는 거의 알려진 바가 없다. 본 연구는 proteasome 저해제인 lactacystin과 HDAC 저해제인 TSA의 병용처리가 세포자멸사 유도에 미치는 영향을 알아보고자 시행되었다. TSA를 전처리한 후 연이어 lactacystin을 처리한 군은 강한 항암효과와 핵의 응축현상을 보였다. Western blot 분석에서는 TSA와 lactacystin의 병용 처리는 Bax/Bcl-2 분율변화와 XIAP 수준의 감소 및 caspase-7의 활성화와 PARP의 분절의 결과를 얻었다. 그리고 더 나아가서 병용처리한 실험군에서 cell cycle arrest는 G2/M기에서 일어났으며, proteasome 활성은 사라졌다. 본 연구는 TSA와 lactacystin의 병용처리에 의해 일어나는 세포자멸사의 기작에 대해 설명하였고, 본 연구의 결과 자료는 생체의 항암치료전략에 도움을 줄 수 있을 거라고 생각한다. Although much information has been accumulated about the synergistic interaction of proteasome inhibitors and HDAC inhibitors to induce apoptosis in a certain type of cells, much less is known currently about the underlying mechanism. This study was undertaken to explore the combination effect of a histone deacetylase inhibitor, TSA, and a proteasome inhibitor, lactacystin, on the induction of apoptosis. Pretreatment of TSA and subsequent treatment of lactacystin showed the strong antitumor activity and nuclear condensation. Western blot assay showed that combination treatment of TSA and lactacystin increased Bax/Bcl-2 ratio and decreased level of XIAP. Activation of caspase-7 and cleavage of PARP were demonstrated after the combination treatment. In combination treatment group, cell cycle arrest was induced at G2/M phase and abolished increase in proteasome activity. This study is elucidating the mechanims whereby targeting apoptotic machineries may help in directing therapeutic strategies.

Expression of vascular endothelial growth factor receptors in tumor and stromal cells of tongue squamous cell carcinoma

Bong-Wook Park,June-Ho Byun,Young-Sool Hah,Deok-Ryong Kim,In-Kyo Chung,Jong-Ryoul Kim,Uk-Kyu Kim,Bong-Soo Park,Gyoo-Cheon Kim 대한구강악안면외과학회 2007 대한구강악안면외과학회지 Vol.33 No.1

This study was to evaluate the expression of vascular endothelial growth factor receptors (VEGFRs) in tumor and stromal cells of tougue squamous cell carcinoma (SCC). We also wanted to characterize the differences, from the angiogenic aspect, between cancer-associated stromal cells and non-malignant stromal cells. Paraffin-embedded tumor specimens from eleven patients with tongue SCCs were studied. Immunohistochemical staining for VEGFR-1,-2, and -3 was performed on the tumor cells, stromal fibroblasts and tumor-associated macrophages of the specimens. The expression of all 3 receptors was detected in the tumor cells themselves of the biopsy specimens. All 3 receptors were also expressed on stromal cells, except that VEGFR-2 was not expressed in stromal fibroblasts. In radical excision specimens, the staining intensity for VEGFR-1, -2 in the tumor cells and VEGFR-1,-3 in the tumor-associated macrophages was significantly lower than that in the biopsy specimens (P < 0.05). By using the general marker of fibroblast and macrophage, 5B5 and CD68, respectively, we performed double immunofluorescence staining for 5B5 and each VEGFR in the stromal fibroblasts and for CD68 and each VEGFR in the tumor-associated macrophages of the radical excision specimens. We used 4 cases of fibroma and 4 cases of chronic inflammation tissue as the controls. It was found that only each marker was expressed in the control group, however, 5B5/VEGFR-1 and 5B5/VEGFR-3 in the stromal fibroblasts, and CD68/VEGFR-1 and CD68/VEGFR-3 in the tumor-associated macrophages were double stained in the radical excision specimens. Although our study used small number of specimens, the results of our study showed that in tongue SCC, in association with the angiogenesis, the stromal cells showed the activated phenotype and this was different from the nonmalignant stromal cells.