본 연구는 착상전 이배체 단위발생 돼지난자를 체외 배양시 우태아혈청 (FBS), 우혈청 알부민 (BSA) 및 상피세포성장인자 (EGF)를 배양액에 첨가하였을 때 배반포, 총 세포수, 세포사멸 및 세포사멸에 관여하는 유전자의 발현을 조사하고자 수행하였다. 0.4% BSA를 배양액에 첨가하였을 때 2세포기 단위발생 난자의 배반포까지의 발달율이 증가되었다(P<0.01). FBS는 배반포의 총세포수를 감소시 켰고 세포사멸을 증가하였다(P<0.01). 그리고 EGF는 BSA가 존재하는 조건하에서 배반포의 총세포수를 증가하였는데 EGF와 BSA가 각각 단독으로 존재할 때는 이런 작용이 없었다. 세포사멸도 이와 이슷한 경향을 보였는데 EGF와 BSA가 각각 존재할 때에는 비처리군과 차이가 없었지만 함께 존재할 때에는 세포사멸을 감소시켰다. RT-PCR의 결과에 의하면 EGF는 BSA가 존재하는 배양액에서 Bcl-xL 유전자의 상대적 발현량을 증가시키고 Bak 유전자의 상대적 발현량에는 영향을 주지 않는 과정을 통하여 세포사멸을 감소시키는것 같다. 반면에 FBS는 Bcl-xL의 발현량을 감소시키고 Bak 유전자의 상대적 발현량을 증가시킨다. 이러한 결과는 세포사멸에 관여하는 유전자의 발현은 배양액의 첨가물에 따라 유의적으로 영향을 받으며, 체외배양시 배아의 초기발달에 관여함을 시사한다.

A case of odontogenic ghost cell tumor (OGCT) with clear cell components occurred in the mandible of a 63-yearold man. The tumor revealed ameloblastomatous type epithelial components together with clusters of ghost cells and dentinoid juxtaposed to the odontogenic epithelium. In addition, some areas of the tumor tissue showed sheets and islands of clear, glycogen-rich epithelial cells, separated by a thin fibrous connective tissue stroma. Both ameloblastic and clear cells showed positive immunoreactivities for cytokeratin 19 and AE1/3. It is not known whether this tumor represents a clear cell differentiation of a preexisting OGCT or a separate and distinct neoplasm derived de novo from odontogenic epithelium. This tumor was preferred the term clear cell odontogenic ghost cell tumor, which captures the clear cell components, one of the tumors most prominent distinguishing features

E6 and E7 as two major oncoproteins of HPV can act together to produce several tumors, providing an evidence for the role HPV in oral squamous cell tumorogenesis. It is worthy to detect E6/E7 mRNA expression of HPV in oral carcinoma. The purpose of this study were to detect HPV mRNA in HNSCC cell lines, and to use these results to confirm oral squamous cell carcinoma. Semi-quantitative RT-PCR method for E7 mRNA expression in HNSCC cell lines should play an important role in detecting the early stage of oral squamous cell carcinoma.

Established SGT cell line from human submandibular gland adenocarcinoma was used to study the TGase expression on a cellular level in vitro. Transglutaminase 2(TGase 2) is assoacitated with apoptosis, GTP binding protein, and cell marix interaction. The role of TGase 2 in salivary gland tumors is not clear yet. The pupose of this study were to examine the TGase expression of SGT cell line compared to other tumor cell lines, and to apply these results to the pathogenesis of salivary gland tumor. TGase enzyme assay of SGT, SCC-15, HN 4 and HeLa tumor cell line was 3 times repeated, and calculated. Immunoslot blot for semiquantitative protein analysis was done. The obtained results were as follows. 1. SGT cell line showed the highest TGase 2 enzyme activity(about 6-16 folds) irrespective of pre or postconfluency. 2. HN 4 cell line showed the highest TGase 1 enzyme activity(about 2-3 folds) irrespective of pre or postconfluency. 3. Under postconfluency TGase 1 induction was not induced, but slightly increased in all tumor cell lines. 4. TGase enzyme activity in all tumor cell lines was accompanied with TGase protein formation. From the aboving results, the higher TGase 2 expression of SGT cell line suggested that they would come from submandibular ductal cells and have a important role in the pathogensis of salivary gland tumors.

Cell adhesion is used as a parameter to evaluate the biocompatibility of dental implant and also affected by the surface form of dental implant. Most study have showed different cell reaction by the composition and the surface morphology of implant. Therefore it is thought that the osteoblastic activity would be affected by the surface roughness and composition of implants. This study was performed to evaluate the biological activity and morphological change of normal human osteoblastic cells(NHost) depending on the variations of implant surfaces. We used grade 2 titanium disks which were being air-blasted with TiO2 50 ㎛, 110 ㎛, 250 ㎛ powder by 3psi compressed air and non-blasted as control. We evaluated and compared morphologic change, adhesion assay, and Ca, P, ALP concentration of NHost in vitro. The obtained results were as follows. 1. In the growth curve, although the growth of experimental groups were lower than that of the group of NHost only, there was no significant difference between each groups. 2. Inverted microscopic findings showed NHosts in early stage of each group were adherant perpendicular to the titanium disk and the multilayered NHosts were attached with various directions after 4 weeks. 3. Scanning electron microscopic(SEM) features showed that NHosts in all groups seemed to be attached multilayered and connected with each process after 2 weeks. 4. NHosts' processes were found by the SEM after one day culture. The cell adhesion of experiment group was higher than that of control group. 110 ㎛(the 3rd group) showed prominant process of NHost on the titanium disk surface. 5. Although the concentrations of Ca, P and ALP were gradually reduced, ANOVA analysis of each groups were partially different, and ANOVA analysis of 4th group were significantly different with others. From the aboving results, NHosts cultured on the titanium disks showed similar morphological change and cell proliferation. There were partially differences in each group except the 4th group, and the 4th group were significantly different with other's in biological activity. We thought that biological activity and adhesion of NHost cell on titanium had been affected by the variation of the titanium surface roughness.

MMPs are catalytic enzymes involved in the degradation of extracellular marix, and associated with invasive growth and metastasis of malignant tumors along with angiogenesis. This study was to evaluate the prognostic significance of VEGF expression and microvessel density(MVD) and the relationship between VEGF expression, MVD and MMPs expression in oral squamous cell carcinomas(OSCC). The materials were from 52 cases of OSCC during a period from 1991 to 2001. Clinicopathologic parameters such as clinical stage, recurrence, histologic grade and invasion pattern were evaluated. Immunohistochemical staining for MMP-2, MMP-9, VEGF and CD34 were performed and statistical analyses between clinicopathologic parameters and VEGF expression and MVD were done. MMP-2, MMP-9 and VEGF expressions were noted in 30(57.7%), 21(40.1%) and 38(73.1%) of 52 cases, respectively. MVD was measurable in 35 cases, and cases with increased MVD more than average were 16(45.7%) of 35 cases. There was no significant relationship between clinicopathologic parameters and VEGF expression or MVD in OSCC, which suggests that VEGF expression and MVD can not be regarded as reliable prognostic factors. Cases with less infiltrative growth pattern showed a tendency of increased MVD, which can explain the possibility that neovascularization might be an early event of tumor invasion. Lack of significant relationship between MMPs, VEGF expressions and MVD might be due to limited number of cases, and positive correlation between MMP-2 and VEGF expression suggests that both factors might be involved in the process of angiogenesis in OSCC.

한우 태아 섬유아세포 유래 체세포복제수정란 이식으로 임신된 수란우가 분만예정일이 도래하였으나 분만징후가 없어 수정란이식 후 272일째에 dexamethasone 20 mg을 근육주사하고 24시간후 PG 25 mg과 estradiol 20 mg을 근육주사하여 분만유도를 실시하였다. dexamethasone 투여후 48시간만 분만징후를 나타내었으며, dexamethasone 투여후 50시간째에 40 kg의 수송아지를 견인에 의해 정상적으로 분만시켰다. 그러

본 연구는 미성숙, 성숙 단계의 돼지 난포란이 유리화 동결에 의한 동결보존이 가능한지를 조사 하고자 실시하였다. 난포란은 세포질 내 지방구를 분극시키기 위해 원심분리를 실시하였고, 미세조작기를 이용하여 지방구를 제거하였다. 돼지 난포란을 CB 처리하여 원심분리 후 지방구를 제거한 지방제거구(Delipated), CB 처리 후 원심분리만 하여 지방구를 분극시킨 원심분리구(Centrifuged), 아무처리도 하지 않은 대조구(Control)를 EM grid

The imbalance between epithelial cell growth and inhibitory factors may cause human epithelial cancer. The dysregulation of growth inhibitory effect of TGF-β1 has been recognized in a variety of carcinomas. This study aimed to investigate the expression of TGF-β1 type II receptors(TβR-II) in the carcinogenesis of oral squamous cell carcinoma(OSCC). Six cell lines established from OSCC in the department of Oral Pathology, Yonsei University College of Dentistry were used. DNA was extracted from harvested cells by phenol-chloroform method. Polymerase chain reaction (PCR) was done with each primer of exon 3, 4, 5, 6 of TβR-II gene. PCR products were inserted to cloning vector (pGEM-T easy vector) and then analyzed to automatic DNA sequencing analyzer. Reverse transcriptase-PCR (RT-PCR) was performed to confirm the mRNA expression of TβR-II gene. Western blotting was performed to detect the expression of the TβR-II protein. As results, a frameshift within a polyadenine region of exon 3 was found in YD-8 cell line. In YD-17 cell line, a missense mutation at codon 238 of exon 4 was found, suggesting the alteration of amino acid from asparagine to aspartic acid. TβR-II mRNA was detected in all cancer cell lines, but it was slightly decreased as compared to that of normal oral mucosal cells. In Western blotting, no TβR-Ⅱ protein was detected in all OSCC cell lines. These results suggested that the altered regulation of TGF-β1 function might play a role in the development of OSCC.

It has been said that amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. System L is a major nutrient transport system responsible for the Na+-independent transport of large neutral amino acids including several essential amino acids. In malignant tumors, a system L transporter L-type amino acid transporter 1(LAT1) is up-regulated to support tumor cell growth. In the present study, we have examined the function of LAT1 and its expression in the KB human oral epidermoid carcinoma cells. RT-PCR, western blot analysis and immunohistochemical analysis have revealed that KB cells express LAT1 in the plasma membrane together with its associating protein 4F2hc, whereas KB cells do not express the other system L isoform LAT2. The uptake of L-[14C]leucine by KB cells is Na+-independent and almost completely inhibited by system L selective inhibitor BCH. The profiles of the inhibition of L-[14C]leucine uptake by amino acids in the KB cells are comparable with those for the LAT1 expressed in Xenopus oocytes. The majority of L-[14C]leucine uptake is, therefore, mediated by LAT1 in the KB cells. These results suggest that the uptakes of neutral amino acids including several essential amino acids in the KB oral epidermoid carcinoma cells mediated by LAT1. In addition, specific inhibition of LAT1 by such agents as BCH in human oral squamous cell carcinomas will be a new rationale for anti-cancer therapy.

칡소의 귀세포를 이용한 핵이식 시에 전기 융합조건이 핵이식란의 발생에 미치는 영향을 알아보고, 칡소의 귀세포와 한우의 태아섬유아세포를 이용한 핵이식란을 이식하여 수태율을 비교하였다. 전기자극 조건에 따른 핵이식 후 융합율은 51∼68%의 범위로 차이는 없었으나, lysis 율은 1.9kv/cm의 10us, 20us에서 각각 0.0과 38.7%, 2.0kv/cm의 10us, 20us에서 각각 2.9와 37.5%, 2.1kv/cm의 10us, 20us에서 각각 21.2와 51.8%을 보였다. 난할율은 1.9kv/cm의 10us, 20us에서 각각 75.8과 69.8%, 2.0kv/cm의 10us, 20us에서 각각 76.9와 68.8%, 2.1kv/cm의 10us, 20us에서 각각 70.5와 68.5로 큰 차이가 없었다. 그러나 배반포 발생율은 1.9kv/cm의 10us, 20us 각각 19.5와 48.6%, 2.0kv/cm의 10us, 20us에서 각각 20.0과 40.9%, 2.1kv/cm의 10us, 20us에서 각각 44.2와 27.0%로 각 조건에서 시간에 따른 차이를 보였다. 핵이식에서 공여세포와 세포질간의 융합율과 lysis율은 전압과 통전시간에 영향을 받으며 전압을 높이는 것이 통전시간을 길게 하는 경우보다 수핵난자의 세포질에 상해를 줄이고 배반포 발생에 유리 하였다. 한우의 태아섬유아세포의 핵이식후 생산된 배반포를 5마리의 수란우에 이식한 결과 두 마리가 임신되었고(40.0%), 칡소의 귀 세포를 핵이식한 후 생산된 배반포를 19마리에 이식한 결과 3마리가 임신되었다(15.8%).

본 연구에서는 한우 태아의 시기별로 35일령, 50일령, 70일령 및 90일령의 fetal fibroblast cell line을 생산하였고, bovine-specific primer와 Y chromosome-specific primer를 이용하여 PCR에 의해 성을 판별하여 각각 암수 2 line의 한우 fetal fibroblast cell line을 확립하였다. 이들 cell line을 계대배양하여 passage number가 10 이상에서 염색체 분석을 실시하였는데 모두에서 80%이상의 세포가 60개의 정상 염색체수의 나타내어 계대배양이 karyotype에 영향을 미치지 않는 것으로 나타났다. Serum starvation과 confluent 배양 방법을 이용하여 Go 상태로 유도되었는지 확인하기 위해 PCNA antibody를 이용하여 Western blotting 분석을 실시하였는데 PCNA 발현이 현저히 감소되는 것을 확인할 수 있었고, 다시 정상 medium으로 환원시켰을 때 세포분열이 재개되어 Go상태로 유도되었음을 확인할 수 있었다. 또한 serum stravation 방법이 conflent한 배양방법보다 PCNA 발현양이 적은 것으로 나타나 좀더 효율적인 Go 상태 세포 주기 조절방법으로 판명되었다.

There is considerable evidence that ionizing radiation (IR) mediates checkpoint control, repair and cell death. In this study, we have used a high density microarray hybridization approach to characterize the transcriptional response of human breast carci