UV and O3 are materials used in the water treatment process, and many studies have been reported to remove organic matters, contaminants, and microorganisms. In this study, we were investigated effects of Chirnomidae (Chironomus flaviplumus, Chironomus riparius), which are contamination indicator species to exposure UV and O3 for the survival rate, body color change and gene expression response. The survival rate of C. flaviplumus exposed to UV decreased to about 70% after 24 hours, and C. riparius about 50%. There was no change in the survival rate of C. flaviplumus exposed to O3, and C. riparius decreased to 95% after 10 minutes of exposure, but there was no change during the subsequent exposure time. In addition, UV and O3 exposure to the two species in body color faded in a time-dependent. In the HSP70 gene expression, C. riparius showed an increase in expression after UV exposure compared to the control group, and a significant difference was shown 12 hours after exposure (P<0.05). C. flaviplumus exposed to O3 showed a relatively low expression compared to the control group, and showed a significant difference at 10 minutes and 1 hour after exposure (P<0.05). These results reported the ecotoxicological effects on Chironomidae according to UV and O3 exposure. Therefore, the results of this study can be used as basic data to understand the effects of UV and O3, which are disinfectants used in water treatment plants, on Chirnomidae entering plants. Key words: Chironomus flaviplumus, Chironomus riparius, UV, O3, acute toxicity, survival

This study was conducted to evaluate in vitro antioxidant activity of goat meat hot water extracts and the changes in apoptosis-related protein expression levels in the cancer cells treated with these extracts. Goat meat hot water extracts were prepared using different cuts of goat meat, including foreleg, hindleg, loin, and rib. Among these extracts, the foreleg and hindleg extracts displayed higher (P<0.05) ABTS radical scavenging activity than the other two extracts. Protein expression levels of BAX, p53, and p21 were not different in the cells treated with the extracts from different cuts, regardless of the cell type. Only p53 expression in HT-29 cells was elevated (P<0.05) after loin extract treatment. These results suggest that antioxidant activity and apoptosis-related effects of goat meat hot water extract varied with cut of meat under in vitro conditions. Because all data was obtained from the in vitro experiment, the ability to generalize conclusions is limited. Additional in vivo studies are necessary.

돌기해삼 Apostichopus japonicus는 주요 양식 대상 무척추동물로서 우리나라 연안 해역에 서 식하고 있다. 본 연구는 방류 방법에 따른 단기간의 생리학적 스트레스 정도를 평가하기 위하 여 heat shock protein 90 (HSP90) 유전자의 발현 변화를 실시간 정량적 중합효소연쇄반응법 으로 조사하였다. 어린 돌기해삼을 비닐봉지에 산소 포장하여 30분간 수송하거나 방류 해역의 간조기에 1시간 공기 중에 노출된 실험군의 HSP90 유전자 발현은 대조군의 HSP90 유전자 발현에 비하여 통계학적으로 유의미하게 증가하였다(수송 후 실험군 p=0.001; 간조기 실험군 p=0.032). 어린 돌기해삼을 방류 후 6시간까지 분석한 결과, 선상에서 씨뿌림 방식으로 방류된 6시간째의 개체 및 호스를 통과하여 수중으로 방류된 2~6시간째의 HSP90 유전자 발현율은 대 조군에 비하여 약간 감소하는 경향을 보였다(씨뿌림 실험군 p=0.069; 호스 방류군 p=0.093). 한 편, 잠수부에 의해 수중에서 방류된 어린 돌기해삼은 방류 후 시간이 경과할수록 HSP90 유전 자 발현율은 증가하는 패턴이 관찰되었다(p=0.061). 이상의 결과는 방류된 어린 돌기해삼의 단기간 스트레스 반응 연구와 효과적인 방류 방법의 개발에 HSP90 유전자 발현이 유용하게 사용될 수 있음을 시사한다.

이 연구는 노화된 흰쥐를 대상으로 규칙적인 저항성 운동에 유산소 운동을 병행하는 훈련을 실시하여 골격근에서 나타나는 혈관신생 관련 단백질 발현의 반응을 관찰하기 위해 수행되었다. 연구의 목적을 위해 자연적으로 노화된 SD계열 흰쥐(20-24개월령, N=18)를 사용하여 통제(CON, n=6), 저항성 운동 (RE, n=6), 저항성+유산소 운동(RE+AE, n=6) 집단으로 구분하였다. 저항성 운동 집단은 실험동물용 사다리를 이용하여 매회 3세트×4회의 운동을 실시하였고 저항성 운동+유산소 운동 집단은 매회 2세트×3회의 사다리 오르기와 추가적인 30분간의 트레드밀 달리기를 수행하였다. 총 8주간의 운동 훈련 종료 후 가자미 근과 장지신근을 적출하여 분석에 사용하였다. 골격근에서 혈관신생 관련 단백질들(HIF-1α, VEGF, FLK-1, Ang-1, Ang-2)의 발현 수준을 분석하기 위해 western blot을 실시하였다. 연구결과, 저항성+유산소 운동 집단에서 가자미근(type I 근육)의 HIF-1α, VEGF, FLK-1, Ang-1, Ang-2 단백질 발현이 통제 집단에 비해 높았으며 저항성 운동만 수행할 경우 HIF-1α, VEGF, Ang-1, Ang-2 단백질 발현이 통제 집단에 비해 높았다. 또한 가자미근에서 저항성+유산소 운동훈련 집단의 Ang-2 to Ang-1 ratio가 저항성 운동 집단에 비해 높아 운동훈련 유형별 차이를 보였다. 한편, 장지신근(type II 근육)에서 HIF-1α는 저항성 운동 훈련에 의해서만 증가된 반면 VEGF와 FLK-1 단백질 발현은 두 훈련 유형 모두에서 증가되었고 운동 훈련 유형별 차이는 관찰되지 않았다. 또한 장지신근의 angiopoieitin 단백질들의 발현은 운동 훈련에 의한 차이가 없었다. 그러므로 노화에서 규칙적인 운동 훈련은 운동 유형에 관계없이 골격근 혈관신생 반응을 유도하며, 특히 저항성 운동에 유산소 운동의 병행은 type I 근조직 유형에서 혈관신생에 대한 추가적인 긍정적 효과를 가질 수 있다.

참다슬기 아가미 조직으로부터 heat shock protein 70 유전자를 분리 · 동정하였다. 참다슬기 HSP70 cDNA의 open reading frame (ORF)는 1,917 bp로 639개의 아미노산을 암호화하여 분자 량은 약 70 kDa으로 예측되었다. 생물정보학 배열분석에 의해 HSP 유전자 기능과 관여되어 있는 3가지 주요 signature motifs와 보존된 도메인을 확인하였다. 계통학적 분석을 통하여 참 다슬기 HSP70 유전자는 왕우렁이 Pomacea canaliculate와 같은 클러스트에 포함된다는 사실을 확인하였다. 수온 및 염분 변화에 따라, 참다슬기 HSP70 mRNA 유전자 레벨은 유의적으로 증 가하였으며(p < 0.05), 이는 외부자극요인을 파악할 있는 분자생물학적 마커로서 활용될 수 있 을 것으로 사료된다.

이 연구는 비만이 심장 조직에서 자가포식 관련 단백질 발현에 미치는 영향을 확인하기 위해 고지방 식이(20주)를 통해 비만을 유도한 후 8주간의 트레드밀 운동을 실시하고, 자가포식의 유도, 형성 그리고 자가포식포와 라이소좀 융합단계를 조절하는 단백질의 발현을 확인하였다. 실험동물(SD rat)은 20주 간의 고지방식이(탄수화물: 20%, 지방: 60%, 단백질: 20%)를 통해 비만을 유도하였으며, 8주간의 트레드 밀 운동(주 5일, 하루 30분, 5분; 8m/min, 5분; 11m/min, 20분; 14m/min)을 실시하였다. 집단 구분은 정상식이 비교군(n=10), 고지방식이 비교군(n=10), 고지방식이 운동군(n=10)으로 구분하였다. 8주간의 트레드밀 운동 실시 전과 후에 경구당부하검사를 실시하여 곡선하 면적(area under the curve; AUC)을 산출 하였으며, 공복시 인슐린 농도와 포도당 농도를 통해 인슐린 저항성 지표인 HOMA-IR과 체중 당 복부지 방량(abdominal visceral fat/Body weight; AVF/BW)를 산출하여 비교하였다. 또한 심장 조직에서 자가포식 관련 단백질을 분석하여 운동 트레이닝의 효과를 검증하였다. 장기간의 고지방식이를 통해 HFD-CON 그룹에서는 비만이 유도되었으며, ND-CON 그룹에 비해 체중, AUC, HOMA-IR, AVF/BW가 증가되는 것으로 나타났다. 하지만 8주간의 트레드밀 운동을 실시한 HFD-TE 그룹에서는 AUC, HOMA-IR, AVF/BW가 개선되는 것으로 나타났다. 체중의 경우, 감소되는 경향은 있었지만, 통계적으로 유의한 차이는 없었다. 자가포식 유도에 관여하는 mTOR와 AMPK는 비만상황에서 모두 감소되었지만, 운동을 통해 증가되는 것으로 나타났다. 자가포식 형성에 관련된 Beclin-1, BNIP3, ATG-7, p62, LC3는 비만상황에서 모두 증가하는 것으로 나타났으며, 운동을 통해 감소되는 것으로 나타났다. 자기포식포와 라이소좀 융합단계 조절하는 Cathepsin L과 LAMP2는 비만상황에서 모두 감소되었으며, 운동을 통해 증가하는 것으로 나타났다. 트레드밀 운동과 같은 신체활동은 대사성 질환에서 나타나는 병리학적 현상을 개선하고 자가포식 과정을 정상적으로 유도하는 것으로 나타났다. 따라서 트레드밀 운동이 심장 관련 질환의 예방 및 치료에 있어 일차적으로 고려해야할 필요성이 있다고 제안한다.

5-fluorouracil (5-FU) is a pyrimidine analog which can work as antineoplastic antimetabolite by blocking thymidylate synthetase conversion of deoxyuridylic acid to thymidylic acid in DNA synthesis. This study is aimed to know the anticancer effect of 5-FU on the expressions of important signaling proteins in KB cells through immunoprecipitation high performance liquid chromatography (IP-HPLC). KB cells were treated with 5 μM 5-FU and cultured for 12, 24, 48, 72, and 96 hours, and followed by IP-HPLC analysis using 32 antisera. 5-FU suppressed the proliferation of KB cells by decreases in the expressions of proliferation-related proteins, Ki-67, PCNA, CDK4, and MPM2 to 82.6%, 92.4%, 95.2%, and 95.9%, respectively, but increases of antiproliferation-related proteins, p16 and p21 to 106.7% and 125.5%, respectively, during 96 hours of experiment. This proliferation reduction was also negatively regulated by cMyc/MAX/MAD network signaling. The cellular protection and survival were consistently arrested by 5-FU treatment in KB cells. The expressions of NFkB, MDR, p-mTOR, and TNFα were decreased to 95.1%, 92.8%, 93.4%, and 90.3% in 48-72 hours, respectively, while cellular stress was increased by upregulation of p38 to 111.3% in 48 hours. And the expressions of pAKT1/2/3, hTERT, and AMPK were also decreased to 93.3%, 97.4%, and 89.3% in 24-48 hours, respectively, while the cellular transformation might be undergone by upregulation of TGF-β1 to 117% until 96 hours. Particularly, 5-FU treatment greatly induced the cellular apoptosis in KB cells by increased expressions of PARP, cPARP, caspase 9, c-caspase 9, caspase 8, and caspase 3 in the lack of p53/BAX and FASL/FAS signaling. The expressions of PARP and c-PARP were increased maximum to 119.2% in 24 hours, and followed by increases of caspase 9, c-caspase 9, caspase 8, and caspase 3 to 111.2%, 125.9%, 108.6%, and 116.3% in 72-96 hours. Therefore, it is presumed that 5-FU induced cellular apoptosis in KB cells may be derived from the overexpression of PARP due to the increased DNA defect caused by 5-FU, which can lead to ATP depletion and subsequent cellular apoptosis.

Coffee is one of the most familiar beverages to modern human adults, but its bio-physiological effect has not been clearly elucidated. It was known that more than one thousand chemicals were included in the ordinary coffee extract. Among them, the caffein and chlorogenic acid (caffeoylquinic acids) are most abundant and have been investigated by many authors so far. In order to know the real cellular effect of whole coffee extract elements, the dialyzed coffee extract (DCE)1) was made to get coffee elements less than 1000 Da molecular weight, which are freely absorable through gastrointestinal tract. It was directly treated in the culture of RAW 264.7 cells, a murine macrophage lineage. RAW 264.7 cells were treated with DCE equivalent to 2.5 cups of coffee (DCE-2.5), DCE-5, and DCE-10 for 12 hours, and their protein extracts were examined by histological observation and immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the proliferation-related proteins depending on the dose of DCE. DCE-2.5 and DCE-5 enhanced the cellular growth of RAW 264.7 cells by increasing the expression of β-actin, PCNA, Ki-67, MPM2, MAX, cMyc, E2F-1, and Rb-1, and by decreasing the expression of MAD and p21. These proliferation-related proteins were rarely affected by DCE-10. DCE-2.5 and DCE-5 induced the cellular proliferation of RAW 264.7 cells by the signaling of E2F-1 and cMyc, respectively, but these cellular effects almost disappeared in DCE-10. Therefore, it was presumed that the low dose of coffee, DCE-2.5 and DCE-5 might be effective for the proliferation of murine macrophages, RAW264.7 cells, contrast to the high dose of coffee, DCE-10. It was also suggested that the low dose of DCE-2.5 and DCE-5 be helpful to increase the innate immunity in vivo by increasing the cell number of macrophages in contrast to the high dose of DCE-10.

The aim of this study was to investigate change of plasminogen activators (PAs) and their inhibitors (PAIs) mRNA and protein expression level by heat stress in porcine endometrial cells. The endometrial epithelial cells were isolated from endometrial epithelium in porcine uterus and cultured in different temperature conditions (38.5 and 41.5℃) for 24 h. Expression of urokinase-type PA (uPA), tissue-type PA (tPA), PA inhibitor-1 (PAI-1) and -2 (PAI-2) mRNA in epithelial cells were analyzed using reverse transcription-PCR and protein levels were measured by immunofluorescence. In result, mRNA expression of uPA, tPA, PAI-1 and PAI-2 were decreased in 41.5℃ than 38.5℃ culture condition, however, significant differences were no detected. uPA, tPA and PAI-2 protein were mainly expressed in nucleus, whereas PAI-1 was distributed in cytoplasm and nucleus. uPA and tPA protein levels were increased by heat stress treatment and significant difference was only detected in tPA level (p<0.05). In contrast, two types of PAIs protein level were decreased in 41.5℃ cultured group compared with 38.5℃ group. In present study, tPA protein expression was upregulated by heat stress in porcine endometrial cells. This result suggest that change of tPA by heat stress may be related to blood flow into uterus and intrauterine microenvironments, and could directly and indirectly influence to reproductive performance in pigs.

Immunoprecipitation-based high performance liquid chromatography (IP-HPLC) is a type of modified enzyme-linked immunosorbent assay (ELISA) that uses protein A/G (or antibody)-conjugated beads instead of the antibody-conjugated wells used in ELISA. In order to determine the fidelity of IP-HPLC, the author used 83 antisera to identify protein expression changes caused by cisplatin treatment in KB human oral cancer cells. KB cells were cultured for 12 or 24 hours with 10 ug/mL cisplatin. The results obtained by IP-HPLC were comparable with published cisplatin data, although ELISA was not conducted in the present study. Cisplatin dominantly reduced the levels of proteins associated with cell proliferation, transcription factors, growth factors, cytoskeletal proteins, and cellular differentiating factors, but on the other hand, apoptosis-related factors, oncogenes, and protective proteins were usually up-regulated, presumably to address cisplatin-induced DNA damage. In particular, cisplatin directly inactivated genomic DNA by down-regulating histone H1 and demethylase and by up-regulating deacetylase. Cisplatin also rapidly induced p53 overexpression and mitochondria-mediated endogenous apoptosis occurred after 12 hours of cisplatin treatment, although this was almost completely replaced by FASL/FAS-mediated exogenous apoptosis after 24 hours. This preliminary study was conducted to investigate the anticancer effect of cisplatin on the KB human oral cancer cells and to determine the fidelity of IP-HPLC data. It was concluded that IP-HPLC is useful for identifying profile changes of genome wide essential proteins and signaling changes of major molecular pathways.

The purpose of this study is to investigate the effects of arabinoxylan rice bran and endurance exercise training on TLR4 mediated protein expression in LPS-treated rats. The results showed that TLR4 as an important protein in the inflammatory response against lipopolysaccharide was shown to be significantly lower in both arabinoxylan supplement with exercise group and exercise group, thus the arabinoxylan rice bran had a higher inhibitory activity than arabinoxylan supplement group. However, NF-κB and MyD88 protein expression was not changed in arabinoxylan supplement with exercise training group, whereas NF-κB significantly decreased in 4 weeks of exercise training group. These results suggest that the supplement of arabinoxylan rice bran with exercise is likely to contribute to inflammation response and the arabinoxylan rice bran can be used as a possible safe alternative to the immunotherapeutic intervention.

Oral squamous cell carcinoma (OSCC), is the most common malignancy of oral cavity, and the sixth most frequently diagnosed cancer worldwide. This tumor type is associated with poor prognosis, and most OSCC patients are diagnosed after the cancer has reached an advanced stage. The over expression of NF-κB p65 has been associated with OSCC progression and lymph node metastasis. Hence, the present study analyzed the expression of NF-κB p65 in OSCC from Korean patients. Immunohistochemistry for NF-κB p65 was performed using 12 normal oral mucosas (NOM), 16 oral leukoplakia (with/without dysplasia), and 58 OSCC patients samples. Immunoreactivity was semi-quantitatively scored and the correlation between the expression of NF-κB p65 and clinicopathological parameters of OSCC patients was analyzed. Immunohistochemical staining revealed that NF-κB p65 expression level increased in oral leukoplakia with dysplasia and OSCC. Moreover, the immunoexpression of NF-κB p65 appeared to be associated with age, recurrence, TNM stage, and lymph node metastasis in OSCC patients (p<0.05). These results indicated that NF-κB p65 can play a role as oncogene in OSCC. Moreover, NF-κB p65 may play an important role in both oral carcinogenesis and OSCC patient outcome. It may be considered as another new malignant biomarker of OSCC.

배큘로바이러스를 이용한 유용단백질 생산량 증대를 위하여 여러 가지 방법이 개발되어지고 있다. 그 중 기존 AcNPV의 다각체 단백질 부분을 이용하여 목적단 백질의 N-말단 부위에 융합 발현함에 따라 목적 단백질의 발현량을 증대시킬수 있 는 다각체 단백질 부분 융합 시스템이 개발되었다. 본 연구에서는 이 시스템의 누에 를 이용한 목적 단백질의 생산 적용성을 확인하기 위하여 기존 보고와 동일한 다각 체 단백질 서열 7가지를 이용하고자 하였다. BmNPV 전이벡터를 이용하여 7가지 다각체 단백질 각 서열과 형광단백질을 가진 각각의 재조합 바이러스를 제작하였 다. 각 융합단백질은 누에를 이용하여 발현을 유도하였으며, 누에의 혈림프와 지방 체를 수거하여 각각 SDS-PAGE 및 Anti-EGFP monoclonal 항체를 이용한 Western blot 분석으로 그 발현을 확인하였다. 그 결과, 각 융합단백질은 형광단백 질 크기인 27 kDa과 다각체 단백질 각 부분을 융합한 크기에서 확인되었다. 융합단 백질은 비융합단백질과의 발현량을 비교하였을 때 발현량이 증대되는 양상을 나 타내었다. 이와 같이, 다각체 단백질 부분 융합을 통한 유용단백질을 생산하는데 누에를 이용함으로써 효율적인 단백질 대량생산에 이용 가능성 및 생산비용 절감 의 이점이 있을 것이라 생각된다.

Odontogenic cysts are classified into inflammatory and developmental origins. The most common representative inflammatory cyst is periapical cyst and the most common representative developmental cyst is dentigerous cyst and cyst which show character of tumor is odontogenic keratocyst and cyst of which cystic epithelial lining cells transform to ameloblastoma is unicystic ameloblastoma. Proliferation studies are needed because of various causes, different clinical and pathologic findings. Recently Ki67 has been generally used as cellular proliferation marker, which is closely related to proliferation. Because Ki67 exists in all the time of cell mitosis stage including G1, S, G2, and M, but disappear in resting phase, G0, it is widely used in the evaluation of cell and tissue proliferation activity. The purpose of this study was to establish clinical therapeutic standard through clinical prognosis associated with Ki67 protein expression because of various causes, different clinical and pathologic findings. Immunohistochemical study was performed in selected each 10 biopsy cases through LSAB reaction and HRP system using anti-Ki67 monoclonal antibody. Ki67 expression was mainly seen in the basal layer of periapical cyst, dentigerous cyst and unicystic ameloblastoma, and in suprabasal layer of odontogenic keratocyat, while positive cells appeared very low frequently in unicystic ameloblastoma. Ki67 expression was mainly observed around inflammatory area. Ki67 expression appeared to be independent on the destruction and recurrence of cystic lesion. Conclusively, high cellular proliferation could not represent destruction and recurrence degree of lesion, but this proliferation might be closely associated with circumstance such as inflammation

곤충(초파리, 모기, 누에)은 해충방제, 유용물질생산, 의학연구 등을 위해 유전자변형 곤충으로 개발되어 왔지만, 아직까지 국내에서 유전자변형 곤충에 대한 환경위해성 평가 등이 거의 실시 되지 못하고 있다. 따라서, 본 연구에서 유전자변형 누에의 환경위해성 평가를 3가지 항목(이동성, 생존능력, 산란 및 부화율)으로 진행하였다. 첫째, 기본 사육 절차에서의 탈출가능성(이동성), 둘째, 사육환경으로부터 탈출시 생존 가능성(8개의 극한 환경조건; 고온, 저온, 건조, 습함, 먹이의 유무), 셋째, 비유전자변형 누에♀ x 비유전자변형 누에♂, 비유전자변형 누에♀ x 유전자변형 누에♂, 유전자변형 누에♀ x 비유전자변형 누에♂, 유전자변형 누에♀ x 유전자변형 누에♂으로부터 나온 산란 및 부화율을 비교 하였다. 유전자변형 누에와 비유전자변형 누에의 이동성은 통계적으로 차이가 없었으며, 산란 및 부화율 또한 통계적 차이가 없었다. 다만, 비유전자변형 암수쌍에서 산란된 알의 부화율보다 유전자변형 누에♀ 와 비유전자변형 누에♂에서 산란된 알의 부화율이 통계적으로 낮은 결과를 보였다. 극한환경에서의 생존율 실험에서 상대적으로 유전자변형 누에가 비유전자변형 누에보다 생존율이 낮았으며, 특히 고온조건의 환경에서 통계적으로 생존율이 낮은 결과를 보였다. 저온 조건의 경우 누에 유충의 동면으로 인해 실험결과를 명확하게 얻을 수 없었다. 유전자변형 누에와 비유전자변형 누에가 일부의 차이를 보였으나 모든 실험에서 유전자변형 누에의 값이 비유전자변형 누에보다 낮게 나타났으며, 결과적으로 이번 연구에서는 유전자변형 누에의 위해성은 비유전자변형 누에보다 적었다.

The molecular mechanisms of the carcinogenesis of oral squamous cell carcinomas (OSCCs) are highly variable and result in different features of tumor progression, i.e., local tissue destruction and metastasis to regional lymph nodes. A case of OSCC arising from proliferative verrucous leukoplakia (PVL) was analyzed for its protein expression profile by immunoprecipitation (IP) – high performance liquid chromatography (IP-HPLC) by using 72 antisera and comparing results with those of KB cells. OSCC arising from PVL showed stronger expressions of proteins associated with cell proliferation (MPM2, PCNA, eiF5A, DHS, DOHH), cell survival (pAKT, MDM2, survivin), matrix proteolysis (elaffin), tumor suppression (p16, p21, PTCH1), the WNT/β-catenin pathway (SHH, WNT1, APC, β-catenin, snail), proinflammation (TNFα), angiogenesis (HIF, CMG2, vWF), and cellular protection (HSP-70, FAK, caveolin) and of oncoproteins (STAT3, 14-3-3, K-RAS, PUMA, PIM1) and growth factors (EGFR, bFGF) than KB cells. On the other hand, KB cells showed stronger expressions of proteins associated with apoptosis (caspase-3, caspase-8, caspase-9, PARP, FAS, FASL, TGase-1, BCL2, BAD, BID, BAK, FLIP), matrix proteolysis (MMP-2, MMP-9), transcription signaling (NFkB, p38, E2F-1, HO-1), and tumor suppression (p53, RB1, PTEN) and of oncoproteins (DMBT1, CEA) and growth factor (TGF-β1, c-erbB2, VEGF) than OSCC arising from PVL. These data indicate the cells of OSCC arising from PVL are more resistant and more robust than KB cells. Furthermore, they suggest the oncogenic signalings of OSCC arising from PVL play important roles in the aggressive growth and rapid tumor metastasis to regional lymph nodes

Oral erythroleukoplakia is characterized by severe dyskeratosis intermingled with multifocal erosive spots on the buccal mucosa, dorsal tongue, and lower lip, etc. A case of oral erythroleukoplakia was diagnosed among 83 cases of common oral leukoplakia since 1997. The pathological examination showed the typical features of leukoplakia with severe epithelial dysplasia, exhibiting dyskeratosis, acanthosis, and basal hyperplasia. The oral erythroleukoplakia was explored in comparison with a representative common oral leukoplakia by the immunohistochemical method using PCNA, β-catenin, EGFR, p53, TNFα, pAKT, and STAT3. Oral erythroleukoplakia showed strong positive reaction of PCNA, p53, EGFR, TNFα, pAKT1 and STAT3 in its spinous layer cells and these reactions were reduced in its basal layer cells, while common oral leukoplakia showed diffusely weak reaction of those proteins. Particularly, β-catenin was positive in the nuclei of some basal and spinous layer cells of oral erythroleukoplakia contrast to the common oral leukoplakia. These findings indicated that the present oral erythroleukoplakia was proliferative with the activation of β-catenin pathway, revealed the dysplastic changes of epithelium by the overexpression of EGFR, p53, and pAKT, and also produced inflammatory reaction through the activation of cytokine-dependent signalings of TNFα and STAT3. These data indicated that the present oral erythroleukoplakia might undergo the early stage of multi-step carcinogenesis via the overexpression of different oncoproteins, especially β-catenine, p53, pAKT, and STAT3.

서양종꿀벌(Apis mellifera)과 동양종꿀벌(Apis cerana)에서 온도 스트레스(4℃, 37℃)에 의한 Cu-Zn superoxide dismutase (SOD1)과 thioredoxin reductase (TrxR) 유전자발현 정도를 비교하였다. 그 결과 두 종 모두 처리 후 5시간까지 SOD1과 TrxR의 발현이 급격히 증가하다 차츰 감소하는 경향을 보였다. 스트레스 성 물질(MV, H2O2) 주입에서는 H2O2보다 MV에 의해 현저히 SOD1과 TrxR의 발 현이 증가하였다. 온도스트레스와 물질주입 스트레스 조건하에서 SOD1과 TrxR 의 효소활성을 측정한 결과, 발현시간보다 좀 더 늦은 시간대에서 최대 활성을 보였 다. 그리고 서양종꿀벌과 동양종꿀벌의 SOD1과 TrxR의 단백질 발현 특성을 구명 하기 위하여 E. coli expression system을 이용하였다. 그 결과 SOD1은 약 16 kDa 정도, TrxR은 약 60 kDa 정도의 위치에서 밴드가 관찰되었으며 항체를 이용한 Western blot 결과에서도 동일한 위치에서 밴드를 detection 하였다.

Internalization and expression of extracellular molecules into cells and tissues is known very important process to biological processes and therapy of various diseases. In this study, we analyzed expression pattern of extracellular molecule after transduction into various human cells. To investigate cellular expression of internalized molecule, we used adenovirus containing green fluorescence protein. After infection of adenovirus into various human cells, the efficiency of intracellular gene expression was assessed with determining GFP expressing cells by fluorescence microscopy or FACS. After one day of adenovirus infection into HepG2 and A549, we observed that GFP expression was low at 10moi but expression levels were increased at 100moi in both cells. But, adenovirus infection into HCT116 showed low expression of GFP at concentrations from 1moi to 100moi. After 2 day infection with adenovirus, GFP expression level at 10moi and 100moi was highly increased in HepG2 and A549 compared with 1 day infection. Especially, GFP expression was significantly increased in HCT116 after 2 days infection. However, GFP expressing SKOV3 cells by adenovirus infection were not found in all the experimental conditions tested. For quantitative analysis of GFP expression of cells by adenovirus infection, we carried out FACS analysis. As a result, GFP was expressed at very low levels at 1moi in all cells used in this experiment. GFP expression slightly increased after increasing moi to 10 in HepG2, HCT116, and A549 cells. By 100moi infection of adenovirus, GFP expression was elevated to 10 fold higher than 10moi in HepG2 and A549 and about 4 fold elevation was observed in HCT116. A549 showed 20 fold higher expression of GFP than SKOV3. We also found that GFP expression by adenovirus infection was the highest in HepG2 cells. Protein expression was enhanced by increasing concentrations or time of adenovirus infection. In these results, GFP expression efficiency of adenoviral gene transduction reveals the highest in HepG2 and lowest in SKOV3 among the cells tested. Taken together, we could confirm that intracellular protein expression efficiency by transduction of extracellular gene was different in various human cells. Our study suggests that the cell types and cellular properties should be carefully examined to enhance expression efficiency of extracelluar molecules in biological research and disease therapy

본 리뷰의 목적은 벼 종자 저장단백질 구조분석 및 발현특성분석 결과 종합화를 통하여 종자형질 개선 등의 실용화연구를 위한 기반구축을 모색하는데 있다. 최근 벼 염색체염기서열완전해독 연구 결과를 이용한 유용형질 유전자 분리 및 실용화 연구가 많이 진행되고 있다. 특히 벼 종자 저장단백질은 인류에게는 주요 영양원으로 사용되어지며 종자 발아시에는 식물체 성장을 위한 질소원으로 사용되어진다. 벼 종자 저장단백질의 분류는 용매에서의 용해도에 따라 약산성 및 알카리 용해성의 glutelin, 알코올 용해성의 prolamin, 염 용해성의 globulin으로 나눈다. 벼 염색체 상에는 11개의 glutelin 유전자와 33개의 prolamin 유전자가 존재하며 prolamin 유전자의 경우 5번 염색체 15 Mb 부위에 15개의 유전자가 위치하였다. 이와 같이 종자저장단백질 유전자들이 동일 염색체 부위에 위치하고 있는 것은 진화학적으로 동일 염색체에서 유래하였거나 유사한 유전자발현 조절영역을 가지고 있음을 의미한다. Globulin 유전자는 5번 염색체에 단일 유전자로 존재하였다. 마이크로어레이를 이용한 종자저장 단백질 관련 유전자의 조직 특이 발현 양상을 분석한 결과 glutelin과 대다수의 prolamin 합성 유전자는 종자배유에서만 발현을 하였으며 소수의 prolamin과 globulin 합성 유전자는 종자배유와 발아종자에서도 발현을 나타내었다. 종자 저장단백질의 프로모터부위를 분리한 후 종자에서의 발현 양상을 분석한 결과 glutelin type C1 프로모터가 종자의 전체 부위에서 발현을 나타내었으며 glutelin type B5와 α-globulin 프로모터가 많은 양의 발현을 나타내었다. 본 리뷰를 통하여 벼 종자 저장단백질의 구조및 발현특성 연구 진행사항을 살펴보았다. 이러한 연구 동향분석은 종자형질 개선 및 물질생산 등의 실용화 연구를 수행하는 연구자들에게 최근의 연구 현황을 제공할 수 있을 것으로 생각된다.