갑상샘 소포샘종과 소포암에서 세포주기 관련단백과 세포자멸사 관련단백 발현의 의의

황준영(Jun Young Hwang),최유신(Yoo Shin Choi),박성준(Sung Jun Park) 대한외과학회 2009 Annals of Surgical Treatment and Research(ASRT) Vol.76 No.1

Purpose: To explore the role of cell cycle regulators and apoptosis regulators in carcinogenesis of thyroid, the expression of cell cycle related proteins (cyclin D1, Ki-67) and apoptosis related proteins (survivin, caspase 3, bcl-2, p53) were investigated in follicular adenoma and follicular carcinoma of thyroid. Methods: The following formalin-fixed paraffin embedded surgical specimens were immunohistochemically stained by avidin-biotin complex method for cyclin D1, Ki-67, survivin, caspase 3, bcl-2, p53; 15 cases of follicular adenoma (FA), 31 cases of minimally invasive follicular carcinoma (MIFC) and 12 cases of widely invasive follicular carcinoma (WIFC). Results: The overexpression of six gene products in follicular neoplasms of thyroid was noted in varying frequency. Among them, increased Ki-67, caspase 3 index and overexpression of bcl-2 were noted in statistically significant, widely invasive follicular carcinoma than that of follicular adenoma and minimally invasive follicular carcinoma. Conclusion: These results suggest that the overexpression of Ki-67, caspase 3, bcl-2 appear to play an important role during follicular carcinogenesis of thyroid. In addition, the overexpression of these proteins is related to the differentiation of MIFC and WIFC. However, further molecular genetic studies are required to determine the interrelationships between the expression of cell cycle related proteins and apoptosis related proteins.

Expression of Kip-related protein 4 gene (KRP4) in response to auxin and cytokinin during growth of Arabidopsis thaliana

( Hye Jeong Cho ),( Hye Kyoung Kwon ),( Myeong Hyeon Wang ) 한국생화학분자생물학회 (구 한국생화학회) 2010 BMB Reports Vol.43 No.4

The cell cycle is regulated by cyclin-dependent kinase (CDK)-cyclin complexes as well as other regulators. We isolated Kip-related protein 4 (KRP4) cDNA that encodes 289 amino acids including six conserved domains. To investigate the expression pattern of KRP4 as well as of other cell cycle-related genes associated with plant hormones, Arabidopsis seedlings were cultured on MS medium containing auxin or cytokinin. All seedlings treated with phytohormones displayed an increased proportion of cells in S phase. A higher proportion of cells in G2 phase was observed in seedlings treated with NAA. RT-PCR confirmed that the expression of KRP4 was decreased after treatment with phytohormones, and that CDKA and D-type cyclin transcription was increased. Additionally, mitotic cyclins were up-regulated by NAA treatment. These results suggest that KRP4 as well as other cell cycle-related genes might contribute to the control of plant growth in response to exogenous hormones. [BMB reports 2010; 43(4): 273-278]

Diethylnitrosamine으로 유도된 쥐의 간암화 과정에서 세포주기 및 세포자멸사 관련 단백 발현

심우정(Woo Jung Sim),김용석(Yong Seok Kim),최유신(Yoo Shin Choi),김범규(Beom Gyu Kim),차성재(Seong Jae Cha),임현묵(Hyun Muck Lim),박언섭(Eon Sub Park) 대한외과학회 2008 Annals of Surgical Treatment and Research(ASRT) Vol.75 No.6

Purpose: To explore the role of cell cycle and apoptosis regulators during hepatocarcinogenesis, the expression of cell cycle-related proteins (cyclin D1 and p27<SUP>kip1</SUP>) and apoptosis-related proteins (p53, survivin, caspase 3). Methods: Sprague-Dawley rats were given 120 ppm diethylnitrosamine (DEN) as a carcinogen and sequentially sacrificed. The expression of cell cycle and apoptotic related proteins were examined by light microscopy and immunohistochemistry. Results: During the DEN-induced hepatocarcinogenesis, sequential histologic changes from preneoplastic lesions (altered hepatic cellular foci, hyperplastic nodules, and hepatocellular adenomas) and ultimately overt hepatocellular carcinomas and metastatic lesions were noted. The cyclin D1 were progressively increased from preneoplastic lesions to hepatocellular carcinomas. However, the p27<SUP>kip1</SUP> and the survivine proteins did not show any other difference with the increasing degree of carcinogenesis. The p53 and caspase 3 proteins were more significantly increased in hepatocellular carcinomas than preneoplastic lesions. The cyclin D1 protein expression did not show any correlation with the expression of p27<SUP>Kip1</SUP> protein, but the p53 expression was related to the expression of survivin and caspase 3. Conclusion: From the above results, over-expression of cyclin D1 plays a role in the early and late stages of hepatocarcinogenesis. In addition p53 and caspase 3 might be useful markers for evaluating the risk of malignant transformation.

Urethane으로 유발된 생쥐 페샘암종 발생과정에서 세포주기 관련인자(Cyclin D1, p21, and p27)에 대한 비소의 효과

임성혁(Sung Hyuk Yim),정지훈(Ji Hoon Jeong),견종만(Jong Man Gyeon),박언섭(Eon Sub Park) 대한약학회 2006 약학회지 Vol.50 No.2

The present study investigated an effect of arsenic trioxide on the urethane-induced lung carcinogenesis in mice. To understand its carcinogenesis, we examined proliferating cell nuclear antigen (PCNA), apoptotic index as well as cell cycle-related proteins (cyclin D l, p21, and p27). Urethane was injected intraperitoneally in ICR mice, and then they were sacrificed at 5, 15, or 25 weeks following treatment of arsenic trioxide. Arsenic trioxide was given with tap water at a concentration of 1 mg/l (low-dose) and 5 mg/l (high-dose) for 25 weeks. During the carcinogenesis, sequential histological changes from hyperplasia to adenomas, and ultimately to overt carcinomas were noted. The development of hyperplasias, adenomas, and carcinomas in the lung were slightly increased by the treatment of low-dose arsenic trioxide. However, there is no correlation between dose and tumor multiplicity. The administration of low-dose arsenic trioxide, significantly increased the tumor size. The proliferative index observed on 5 weeks after significantly increased. Cyclin D1 and p21 protein, cell cycle related proteins, were more significantly increased in hyperplasia and adenoma in low dose arsenic treated group than urethane alone group. The p27 protein expression did not show any significantly changes with arsenic treated or untreated group. Low dose exposure to arsenic trioxide resulted in increased expression of cyclin D1 and p21 protein. The present results indicate that low-dose treatment of arsenic trioxide, but not high dose of it, partly modulate the cellular proliferation, cyclin D1, and p21 protein expression, and that this effect may contribute to accelerated development of lung adenocarcinomas in urethane-induced mice.

Mechanism Underlying NaF-Induced Apoptosis in Human Oral Squamous Cell Carcinoma

Young-Joo Hur,Do-Kyun Kim,Seung-Eun Lee,In-Ryoung Kim,Na-Young Jeong,Ji-Young Kim,Bong-Soo Park KOREAN ACADAMY OF ORAL BIOLOGY 2010 International Journal of Oral Biology Vol.35 No.2

Few studies have evaluated the apoptosis-inducing efficacy of NaF on cancer cells in vitro but there has been no previous investigation of the apoptotic effects of NaF on human oral squamous cell carcinoma cells. In this study, we have investigated the mechanisms underlying the apoptotic response to NaF treatment in the YD9 human squamous cell carcinoma cell line. The viability of YD9 cells and their growth inhibition were assessed by MTT and clonogenic assays, respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to detect apoptosis. YD9 cells were treated with NaF, and western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, and MMP and proteasome activity assays were performed sequentially. The NaF treatment resulted in a time- and dose-dependent decrease in YD9 cell viability, a dose-dependent inhibition of cell growth, and the induction of apoptotic cell death. The apoptotic response of these cells was manifested by nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, a significant shift of the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase- 3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF treatment resulted in the downregulation of G1 cell cyclerelated proteins, and upregulation of p53 and the Cdk inhibitor p27 KIP1 . Taken collectively, our present findings demonstrate that NaF strongly inhibits YD9 cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via mitochondrial and caspase pathways.

Mechanism underlying Chios gum mastic-induced apoptosis on SCC25 human tongue squamous cell carcinoma cell line

Lee, Seung-Eun,Hur, Young-Joo,Kim, In-Ryoung,Kwak, Hyun-Ho,Kim, Gyoo-Cheon,Shin, Sang-Hun,Kim, Chul-Hoon,Park, Bong-Soo KOREAN ACADAMY OF ORAL BIOLOGY 2009 International Journal of Oral Biology Vol.34 No.2

Chios gum mastic (CGM) is a resin produced from the stem and leaves of Pistiacia lentiscus L var chia, a plant which grows only on Chios Island in Greece. CGM has been used for many centuries as a dietary supplement and folk medicine for stomach and duodenal ulcers in many Mediterranean countries and is known also to induce cell cycle arrest and apoptosis in some cancer cells. In this study, we further investigated the induction and mechanisms underlying the apoptotic response to CGM treatment in the SCC25 human tongue squamous cell carcinoma cell line. The viability of SCC25 cells, human normal keratinocytes(HaCaT cells) and human gingival fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay, respectively. Staining with Hoechst and hemacolor dyes and TUNEL assays were employed to detect SCC25 cells undergoing apoptosis. SCC25 cells were treated with CGM, and this was followed by western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity analyses. CGM treatment of SCC25 cells was found to result in a time- and dose-dependent decrease in cell viability, a dose-dependent inhibition of cell growth, and apoptotic cell death. Interestingly, CGM showed a remarkable level of cytotoxicity in SCC25 cells but not in normal cells. Tested SCC25 cells also showed several lines of apoptotic manifestation. Taken together, our present findings demonstrate that CGM strongly inhibits cell proliferation by modulating the expression of G1 cell cycle-related proteins and induces apoptosis via the proteasome, mitochondria and caspase cascades in SCC25 cells.

극 저주파 전자기장이 N-bis(2-hydroxypropyl)nitrosamine과 Sulfadimethoxine에 의해 유발된 갑상샘 암에 미치는 영향

이수환(Soo Hwan Lee),박용진(Young Jin Park),박언섭(Eon Sub Park),김용석(Young Seok Kim),최유신(Yoo Shin Choi),김범규(Beom Gyu Kim),박성준(Sung Jun Park),정세민(Se Min Chong) 대한외과학회 2009 Annals of Surgical Treatment and Research(ASRT) Vol.77 No.3

Purpose: Long-term exposure to extremely low-frequency (60 ㎐) electromagnetic fields (ELF-EMF) raises the questions of the induction of biological effects including tumorigenesis. One mechanism through which ELF-MFS could influence neoplastic development is the imbalance of cellular proliferation and cell apoptosis. The present study investigated the effect of ELF-EMF on chemically-induced thyroid carcinogenesis in a rat. Methods: We examined cellular proliferation index measured by anti-Ki-67 antigen, apoptosis, apoptosis related proteins such as caspase 3 and p53, and cell cycle-related proteins (cyclin D1 and p21<SUP>WAF1</SUP>/Cip1). Forty Male F344 rats received a subcutaneous N-bis(2-hydroxypropyl)nitrosamine (DHPN, 2,800 ㎎/㎏) injection, and 1 week later were allowed free access to drinking water containing sulfadimethoxine (0.1%) for 12 weeks. Twenty rats were exposed by ELF-EMF. During the carcinogenesis, sequential histological changes from hyperplasia, adenoma, and ultimately to overt carcinomas were noted. Results: The exposure group of ELF-EMF, significantly increases the number size of carcinomas. Also, the proliferative and apoptotic indices were significantly increased in the ELF-EMF exposure group than in the control group. The caspase 3 protein expression did not show any significant changes between ELF-EMF group and control group. The p53 protein was not detected in both ELF-EMF exposure and control group. Among the cell cycle related proteins, cyclin D1, not p21<SUP>WAF1</SUP>/Cip1, was significantly increased in adenomas and carcinomas in ELF-EMF exposure group compared with the control group. Conclusion: Exposure of ELF-EMF effects on chemically-induced rat thyroid carcinogenesis as results of altered increase of cellular proliferation, apoptosis, and cyclin D1 expression.

The Effect of Environmental Micropollutant (DEET) on the Expression of Cell Cycle and Apoptosis Regulatory Proteins in Human Cells

김인수,Xianghao Ren,장진수,Jin Wook Lee,유혜원,김성조,Jung Sun Heo,Am Jang,한호재 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.2

N,N-diethyl-m-toluamide (DEET) is an insect repellent used worldwide, and a common micropollutant in aquatic environments. However, few studies have addressed the molecular mechanism of DEET toxicity and its effects on cell growth and apoptosis. The purpose of this study was to investigate the effect of DEET on the expression of the cell cycle and apoptosis regulatory proteins in human BE(2)-M17 cells. The results showed that DEET significantly decreased the cell viability (40.6 ~ 68.9% of control)at concentrations of 500 ~ 4,000 mg/L. Also, DEET significantly decreased the expressions of CDK 2, CDK 4, and cyclin D1 (3.9 ~ 86.6% of control), at concentrations of 50~ 400 mg/L but from 100 mg/L for cyclin E. Furthermore,DEET significantly increased the expression of caspase-3(223.1 ~ 1,770.6% of control), but significantly decreased Bcl-2 expression (46.1 ~ 86.3% of control) at all concentrations tested. In conclusion, DEET partially affected the expression of CDK/cyclin molecules, but fully affected the expressions of caspase-3 and Bcl-2 in BE(2)-M17 cells.