농촌진흥청 국립원예특작과학원에서는 2017년에 절화 수명이 길고 수량이 많은 연한 핑크색의 스프레이 장미 ‘Pink Shine’ 을 육성하였다. 모본은 ‘Fire Flash’로 붉은 복색의 스프레이 장미이며, 부본은 ‘Pink Charm’으로 핑크색이며 흰가루병에 강하다. 이 두 품종을 2012년 인공교배하여 이듬해인 2013년 1월에 파종, 9cm 포트 묘에 정식하여 관능 평가 실시 후 도태시켜 39개체 의 실생을 얻었다. 이후 화형, 화색, 꽃잎 수, 절화수량, 병 저항성 등을 고려하여 2015년까지 5개체를 선발하여 유사 품종인 ‘Missha’를 대조로 하여 2017년까지 3차에 걸친 특성 검정을 실시하였다. 그 결과 가장 우수한 ‘원교 D1-325’를 최종선발하여 ‘Pink Shine’으로 명명 후 2018년 3월 22일 품종보호출원(제 2018-212호)하여 2019년 6월 21일에 품종보호권(제7786호)이 등록되었다. 화색은 연한 핑크색(RHS, R36D)이며 잎의 색은 녹색(RHS, G137A)으로 대조 품종 ‘Missha’와 동일하였다. 꽃잎 수는 67.8개, 화폭 5.4cm, 화고 3.2cm로 ‘Missha’보다 컸으며 평방미터당 연간 절화수량은 131본, 절화수명은 15.3일로 ‘Missha’ 보다 우수하였다.

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

This study were to perform for verifying the activation areas in the human's brain during mastication by using functional-MRI (f-MRI) device on the basis of hypothesis regarding anatomical-physiological parts of brain processing the information of motor and sensory function, and to perform further more for a providing basic provisional foundation about diagnosis, treatment and prognosis of abnormal occlusion as applying functional MRI. Generally healthy 10 volunteers who have a normal occlusion were selected. The half of members of volunteers was female. Age distributions were approximately alike. Before taking a f-MRI, sufficient practice was carried out as strict standards and made volunteers be not sensible to sweet taste of gum through chewing gum for 30 minutes before taking a f-MRI. Functional images for all volunteers were firstly obtained, and then anatomical images were next. The functional images consisted of echo-planar image volumes which were sensitive to BOLD (blood oxygenation level-dependent) contrast in axial orientation. The volume covered the whole brain with a 64×64 matrix and 42 slices. Images with 64 volumes were acquired under periodic mastication. The orofacial sensorimotor cortex was primary responsible cerebral part during mastication and insula. And also supplementary motor area and cerebellum in brain were intimately connected with mastication. Other numerous anatomical parts of brain were activated in each volunteer during mastication, but there was no statistical significance in this experiment. Differences according to gender and age were no significance in this study. The f-MRI device showed the accurate and detailed image in activation area of brain through valuable device. It suggested that f-MRI might be helpful to establish the basis of funtional standard occlusion depend on activation area of brain.

The rise of medical knowledge and awareness of the importance of dental and stomatognathic system increase the patients who visit dental emergency room. The chief complaints of patients who visited the emergency room varies from a pain, trauma, hemorrhag. The purpose of this study was to classify dental emergency patients by chief complaint and to analyze in indiviual group and to provide more effective emergency dental care. This study was carried out with 1129 patients visiting emergency room of the dankook university dental hospital from 7/2011 to 6/2012. Dental emergency patients was classified trauma, pain, hemorrhage group by chief complaint and studied mothly, the day of the week, time, age distribution and cause of the individual group. The chief complaint of visiting emergency room were trauma 660 people (58.5%), pain 347 people (30.7%), hemorrhage 96 (8.5%), other 26 patients (2.3%). The monthly distribution was observed in May (12.8%), March (10.5%), September (10.2%). The trauma patients were frequent in the spring and early summer but painful patients visited in September (12.7%) and May (11.8%) in March (11.2%). The peak age group was 20 to 29 years(20.9%), followed by 0 to 9 years(19.4%), 40 to 49years (15.2%,). In trauma group the peak age was under the age of 10 (31.7%), followed by 10 to 19 years (18.8%), 20 to 29 years (17.1%) . However, in pain group, peak age was 20 to 29 years (26.8%) followed by 40 to 49 years(21.3%), 30 to 39 years (19.6 %). The most common cause of trauma were subluxation(16.5%), laceration13.7%), uncomplicated crown fracture(12.05%) and in pain group was pulpal origin(46.1%), followed by periodontal origin( 20.7%), post op pain(8.9%). Undefined pain or neuralgia were 7.9%. The most common cause of hemorrhage was post extraction( 66.7%), post operation(16.7%), spontaneous bleeding due to periodontitis(12.5%). In conclusion, the trauma, pain were different in monthly distribution and the peak age of patients. Dental emergeny doctor should understand pattern of indiviual emergency group and perform proper diagnosis and treatment for more effective emergency care.

Recently, extensive research has been performed in the field of orthopedic medicine to develop cell-based therapies for the restoration of injured bone tissue. But there has been rarely reported about rehabilitaton of oral and maxillofacial bone defect using self-derived osteoblasts. Normal human osteoblast cell(NHost) was previously established into marrow-derived human mesenchymal stem cells for their capacity to proliferate and differentiate into osteoblasts under various culture conditions. The purpose of this study was to examine proliferation and differentiation of NHosts effected by growth factors with ALP activity and RT-PCR. After NHosts were cultured under basal and osteogenic medium at 37℃ and 5% CO2, they were analyzed by ALP activity and RT-PCR. BMP-2 under osteogenic medium decreased growth rate of NHosts compared to under osteogenic medium. BMP-2 under osteogenic medium induced osteoblastic differentiation in NHosts by increased ALP activity. The differentiating capacity of NHosts under osteogenic medium showed that NHosts expressed higher mRNA expression levels of OSX and OCN, while that of RUNX2 decreased after BMP-2 treatment. It suggested that NHosts having characteristics of osteoprecursor cells might be more advanced in their osteogenesis development by BMP-2, making NHosts an interesting biological tool for treatment of skeletal defects and diseases of oral and maxillofacial bone.

Several recent studies have detected genetic and cytogenetic alterations in epithelial odontogenic tumors. However, the detailed mechanisms of oncogenesis, cytodifferention, and tumor progression remain unknown. p63 as p53 homolog gene has been identified at loci 3q27-29. The p53 signaling cascade has an important role in oncogenesis or cyto- differentiation of odontogenic epithelium. Recently, several syndromes associated with p63 gene mutations have shown to include various tooth abnormalities of both the primary and permanent dentition. But little is known about p63 expression in odontogenic tumors, especially ameloblastomas. The purpose of this study were to examine various expression of p63 in ameloblastomas by immunohistochemistry and to clarify the possible biological role of p63 in ameloblastomas. 15 specimens including 6 follicular, 4 plexiform, 3 acanthomatous, and 2 granular cell types were fixed in 10% neutral formalin. 4um thick sections were used for routine H&E and immunohistochemical examinations. After immuno- histochemical satining, they were examined at a final magnification of 400X. For each case a minimum of 1000 nuclei located in the central and peripheral layers were counted in up to 10 consecutive microscopic fields per case. The immunoreactive cells were evaluated semiquantitatively. Immunoreactivity for p63 in all the types of ameloblastomas was higher in peripheral neoplastic cells than in central neoplastic cells. Keratinizing cells in acanthomatous ameloblastoma and granular cells in granular cell ameloblastoma showed markedly decreased reactivity for p63 in acanthomatous and granular cell ameloblastoma. Labelling index of acanthomatous, plexiform, and granular cell type was 86±11%, 81±17% and 83±15% in peripheral area while 88±14%, 82±11% and 76±10% in central area, respectively. Labelling index of follicular type was 17±4% in peripheral area while 21±3% in central area. There was no significant relationship between plexiform, acanthomaous, and granular cell type, while significant relationships between follicular and acanthomatous type, between plexiform and follicular type, and between granular cell and follicular type, respectively. It suggested that p63 expression could paly an important role in the pathogenesis of ameloblastomas. Morever plexiform, acanthomatous, and granular cell type would show more aggressive proliferative potentiality than follicular type.

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 epithleial lining cells transform to ameloblastoma is unicystic ameloblastoma. About ten years ago p63 protein that are closely related to p53 protein was found. Authors studied about comparative pattern of expression of p63 protein in periapical cyst, dentigerous cysts, odontogenic kertocysts and unicystic ameloblastomas. Authors selected 10 cases for every four types of cyst and performed immunohistochemical staining by using monoclonal antibody about p63 protein, LSAB(labelled streptoavidin biotin) reactant and HRP(horse raish peroxidase) system. Positive cells about p63 protein were expressed at basal layer of cystic lining epithelium in periapical cysts, odontogenic keratocysts and unicytic ameloblastomas. On the contrary, in dentigerous cysts positive cells were expressed at surfce layer. Perapical cysts and odontogenic keratocysts showed significantly high values of labelling indices.(periapical cyst:72.49%, odontogenic keratocyst:64.72%, dentigerous cyst:8.94%, unicystic ameloblastoma: 5.25%) Odontogenic keratocyst showed the most strong staining intensity and the second was periapical cyst, the third was dentigerous cyst, and lastly unicystic ameloblastoma. Conclusively cause that the positive cells appeared at surface layer in dentigerous cyst reflected the position of epithelium to the enamel, and labelling indices of p63 protein were closely related to proliferative capacity and intensity of expression closely related to the labelling index and thus labelling index was also closely related to proliferative capacity of cystic lining epithelium.

Ameloblastomas are benign odontogenic tumor and the most common neoplasm in jaws and they have locally invasive property and high recurrence rate. Four typical subtypes ameloblastomas are plexiform, follicular, granular cell and acanthomatous type, but their developmental states during tumorigenesis are uncertain. And thus authors studied about developing states of four types of ameloblastomas by immunohistochemical staining for cytokeratin 8/18 which was an intermediate filament of epithelial cell origin and for vimentin which was an intermediate filament of mesenchymal cell origin, and then by comparative analyses of the results. Authors selected seven cases for every four types of ameloblastomas, and then performed immunohistochemcial staining for cytkeratin 8/18 and vimentin to all selected specimen by using monoclonal antibodies about cytoleratin 8/18 and vimentin, LSAB(Labelled StreptoAvidin Biotin) reactant and HRP(Horse Radish Peroxidase) system. Labelling indices of cytokeratin 8/18 of plexiform and follicular types of ameloblastomas were significantly high values in the group of ameloblast-like cells and labelling indices of cytokeratin 8/18 of all types of ameloblastoma were high values in the group of transformed cells, but their differences were not significant. Labelling index of vimentin of plexiform ameloblastoma was significantly high value in the group of ameloblast-like cells and others showed comparatively lower values. Labelling index of vimentin of granular cell type of ameloblastoma in the group of transformed cells was significantly high value and others showed comparatively lower values. Consequently the most primitive form of ameloblastoma was plexiform, and more differenciated form was follicular type and granular cell type and acanthomatous type were most differenciated form of ameloblastomas

Urokinase-type plasminogen activator(uPA) bound to urokinase plasminogen activator receptor(uPAR) expression is strongly correlated with the metastatic potential of various tumors by enhancing ECM degradation through plasminogen and matrix metallopreotease activation. But expression of uPA/uPAR in human malignant salivary gland tumors has been rarely reported. The purpose of this study were to investigate mRNA expression and cytologic concentration of uPAR in SGT cell line compared to various cancer cell lines by RT-PCR and ELISA method, and to study migration and adhesion assay. These results would be to apply the pathogenesis and prognosis of malignant salivary gland tumors. All the cell lines(SGT, HN 4, SCC 25, and HeLA) were cultured under DMEM with 10% FBS at 37℃ in a 5% CO2 incubator. We studied a possible association between mRNA expression and cytosolic concentrations of uPAR in SGT cell line compared to various cancer cell lines using RT-PCR and an enzyme-linked immunoassay( ELISA) method. And also cell adhesion and migration assay were done in all the cell lines. In migration assay SGT cell line was about 2.5-4 folds higher than another cell lines. In adhesion assay SGT cell line was about 1.1-2 folds higher than another cell lines. uPAR cytosolic concentrations of SGT cell line was about 3.4-10 folds by ELISA, while mRNA expression was about 2.5-5 folds by RT-PCR. Oral Scc cell lines showed the lowest value. uPAR protein and mRNA expression were correlated with migration and adhesion assay. From the aboving results, high cytosolic concentrations and mRNA expression of uPAR were correlated with migration and adhesion assay. It suggested that this might be a specific marker for malignant potential of SGT cell line and would be contributed to pathogenesis and prognosis of human salivary gland adenocarcinoma

Plasminogen activator(PA) system such as urokinase plasminogen activator(uPA), urokinase PA Receptor(uPAR), tissue, tissue PA, and PA inhibitor-1&2(PAI-1&2) play a role in tumor invasion, metastasis, and proliferation. It is interested that these factors in patients with primary oral squamous cell carcinoma(Oral SCC) will be evaluated and correlated with clinicopathologic variables. Recently, these expression of primary oral SCC has been restricted to clinical or immunohistochemical study such in vivo study. The purpose of this study were to investigate the mRNA expression and cytologic concentration of uPA, uPAr, tPA, and PAI-1,2 in oral SCC cell lines compared to NHOK and to apply these results to evaluate early detection biomarkers of oral SCC in future. All the cell lines(NHOK, HN 4 and SCC 25) were cultured under KBM bullet kit at 37℃ in a 5% CO2 incubator. We studied a possible association between mRNA expression and cytosolic concentrations of uPA, uPAR, tPA, and PAI-1,2 in oral SCC cell lines compared to NHOK using RT-PCR and an enzyme-linked immunoassay(ELISA) method. uPA mRNA expression was about 5-6 folds, while uPAR was a bout 3 f olds, and PAI-1 was about 1 .5-1.6 f olds. PAI-2 was a bout1.2 -1.3 f olds t han that o f NHOK, w hile t PA w as l ower t han that of NHOK. uPA cytosolic concentrations was about 15-19 folds, while uPAR was about 8 folds, and PAI-1 was about 3-4.5 folds. PAI-2 was about 2 folds than that of NHOK, while tPA was lower than that of NHOK. Both uPA, uPAR, and PAI-1,2 cytologic concentrations were correlated with mRNA expression of oral SCC cell lines. From the aboving results, high cytosolic concentrations of uPA, uPAR, and PAI-1 & 2 were correlated with mRNA expression. It suggested that these might be specific markers for oral SCC cell lines and these results would be contributed to evaluate early detection biomarkers for human oral squamous cell carcinoma.

The aim of this study is to find out histomorphologic change and cellular activity of condyle resulted from unilateral mastication by comparison of cell proliferation and apoptosis activity. 30 adult rats were dived to 15 experimental group and 15 control group randomly. Right upper and l ower molars were gently extracted in experimental group, to make unilateral mastication environment. All subjects were sacrificed at 1 week, 2 weeks and 4 weeks by chloroform, and their tissues were prepare to observation. Streptovidin-biotin system for BrdU stanning, was used to determine cellular proliferative activity. TUNEL method was used to determine apoptotic activity. The result for cellular activity was recorded at both of anterior portion and posterior portion of condyle. Hematoxylin and Eosin stanning was used for histiomorphological change. The results were as follows. There were more change in superficial layer than deep layer of condyle in cellular activity. In anterior portion of condyle cartilage, cellular proliferative activity of experimental group was lower than control group and apoptotic activity of experimental group was higher than control group. And apoptotic activity of extracted side in experimental group is the most. In posterior portion of condyle cartilage, cellular proliferative activity of extracted side in experimental group was higher than non-extracted side and control group, And apoptotic activity of extracted side in experimental group was the low. As a result of histomorphological change, there was hyperplasia in posterior region o f extracted side c ondyle i n experimental g roup, but t here was n o change i n unextracted side i n experimental group. There was histomorphological hyperplasia in posterior condyle of experimental group as results of high cellular proliferative activity. There was mainly apoptotic change of anterior portion condyle in experimental group. But there was no histomorphologic change. In other words, there was hyperplasia by increasing of cellular proliferative activity in posterior portion of nonfunctional side condyle. In functional side condyle, there was no histomorphological change in functional condyle, but there was change in cellular activity.

Tumor cell biological factors, such as urokinase plasminogen activator(uPA) and its inhibitor plasminogen activator inhibitor- 1(PAI-1) play a role in tumor invasion, metastasis, and proliferation. These factors in patients with primary oral squamous cell carcinoma(Oral SCC) will be evaluated and correlated with clinicopathologic variables. However, relatively rarely has been known in oral squamous cell carcinoma in vivo and in vitro study . The purpose of this study were to investigate the protein expression of uPA and PAI-1 in oral SCC cell lines cell line compared to NHOK and to study migration and adhesion assay. All the cell lines were cultured under KBM bullet kit at 37℃ in a 5% CO2 incubator. We studied a possible association between cytosolic uPA and PA-1 concentrations in oral SCC cell line compared to NHOK using an enzyme-linked immunoassay(ELISA). Cell adhesion and migration assay were done in all the cell l ines. In migration assay oral SCC cell lines were about 70 folds higher than NHOK. In adhesion assay oral SCC cell line were about 7-12 folds higher than NHOK. uPA cy tosolic concentrations was about 15-19 folds and PAI-1 was 3 to 4.5 folds than that of NHOK. Both uPA and PAI-1 concentrations were correlated with migration and adhesion assay. High cytosolic concentrations o f uPA and PAI-1 were correlated with migration and adhesion assay . It suggested that these markers might be specific for oral SCC cell line and these results would be contributed to treatment and prognosis of human oral squamous cell carcinoma.

The regeneration of periodontium is the goal of periodontal therapy. Many periodontologists try to achieve this goal by using guided tissue regeneration(GTR) method. However, these procedures always include several disadvantages. Recombinant human bone morphogenetic protein-2 (rhBMP-2) stimulated ectopic bone formation when it was implanted in rat muscles with insoluble bone matrix by differentiating muscle cells into chondrocytes and osteoblasts. The purpose of this study was to evaluate the osteoinductive potential of the mixture of rhBMP-2 (5 μg/ml) and heparin (0.25 or 25 μg/ml ) at the critically sized rabbit calvarial defects. And this study aimed to reveal that heparin also acts to enhance the bone forming activity of rhBMP-2. The 12 rabbits (4-month-old; NewZealand White) were used in the present study. 5 μg/ml of rhBMP-2 and 0, 0.25 or 25 μg/ml of heparin were mixed and blotted into anorganic bovine bone and filled cranial defects. The animals were sacrificed following a time schedule (1, 3, and 6 weeks). Sections were made in 7 μm thicknesses, stained with H&E and Masson's trichrome method, and examined under a light microscope. The differences among each obtained percent value were evaluated by one-way analysis of variance. A p value of p<0.05 was considered statistically significant and an ANOVA test was performed to verify significant differences. To adjust for multiple comparisons when one-way analysis of variance showed a significant difference between groups (p<0.05), Scheffe`s post hoc test was used to identify which group differences accounted for the significant p-value. In control group, osteoinduction was not outstanding, however, in experimental groups, osteoinduction was significantly outstanding, and as the concentration of heparin mixed with rhBMP-2 increased, osteoinduction was increased. Mixtures of rhBMP-2 and heparin affect bone formation at initial bone formation, but that effect disappeared following a time lapse.

Bone morphogenetic proteins (BMPs) are known to promote osteogenesis, and clinical trials are currently underway evaluating the ability of BMPs to promote bone formation in grafting procedures and fracture healing. Some studies, have independently reported that sulfated polysaccharides particularly heparin, enhance the osteoblastic differentiation induced by BMPs in vitro, and another study demonstrated that heparin enhanced the bone formation induced by BMP‐2 in vivo. This study was performed to examine adipose stem cell responses to rhBMP‐2 alone and rhBMP‐2 with heparin at 0.25, and 25 μg/㎖ concentrations, respectively, in culture media. Adipose stem cells were cultured for 2, 4, and 8 days toward the osteoblastic differentiation in rhBMP‐2 alone and rhBMP‐2 with heparin at 0.25, and 25 μg/㎖ concentrations, respectively, in culture media. Verification of the stem cell lineage was performed in two ways. The first method was a continuous sequential culture until 5th generation. The second method was using monoclonal antibodies for STRO‐1 and CD 90. Naphthol AS phosphate‐fast blue BB staining for alkaline phosphatase was used for verifying osteoblastic differentiation because Alkaline phosphatase activity had been used as an osteoblastic differentiation marker and degree of osteoblastic activity. Alizarin red staining was also used as an osteoblastic differentiation marker because it quantifies the calcium levels in cells or tissues. During the 5th generation culture, cultured cells actively proliferated, and these cultured cells showed a positive reaction to STRO‐1 and CD90 cell surface molecules. Naphthol AS phosphate‐fast blue BB staining and Alizarin red staining were positive in most samples of each group at 2, and 4 days and positive reaction was proportioned to degree of morphological differentiation. In the concentration of 25 μg/ml of heparin, the ALP activity was highest at the 2nd day in the culture, and then the activities of ALP were decreased significantly at 4, and 8 days. The ALP activity was greatest at the 4th day of the culture, and then decreased significantly at the 8th day in 0 μ g/ml and 0.25 μg/ml of heparin concentrations, Adipose stem cells could be differentiated in rhBMP‐2 in culture media, and the addition of heparin to BMP‐2 promoted differentiation of osteoblasts. Moreover, morphological differentiation was associated with the activity of osteoblasts. This study was shown that, when heparin concentration increases, the early differentiation of the cells was brought about, but the early differentiated cells were rapidly progressed to degenerative changes

The purpose of this study was to evaluate the biological characteristic of deproteinized freeze dried bovine bone(DFDBB) through grafting to maxillary sinus as following time lapsed. Nine patients who were needed of sinus elevation procedure because of severe resorption of maxillary edentulous alveolar bone were selected. patients were divided into three group. Firstly sinus lifting procedure was performed and then the implantation procedure was performed after 6 months in first group, 12 months in second group and 18 months in third group and simutaneously tissues of sinus were obtained by trephine. 18 sections are made from every obtained tissue. 9 sections were stained by Masson's trichrome method and were taken a photo at 100 times of magnification. Relative area of newly formed bone were obtained by IPTK(image processing tool kit) version 5.0 program and mean values and standard deviations were produced from obtained data by using SPSS version 17 program and significance tests were conducted by ANOVA method. This study revealed that DFDBB stimulated new bone formation in maxillary sinus and did not have osteoinductive capacity but osteoconductive capacity, and DFDBB was exceedingly slowly resorbed.

The origin of squamous cell components in salivary gland tumor has been not yet clarified in detail. The squamous cell differentiation from adenocarcinoma has been reported in various carcinoma by HPV transfection in vitro. The adenocarcinoma cells adjacent to the squamous cell carcinoma components were positive for HPV. This is thought to indicate that after adenocarcinoma cells are transfected with HPV, they undergo morphological changes, and that squamous cell differentiation follows. The purpose of this study were to examine the effects of HPV-16 E6/E7 gene transfection into SGT cell line from human salivary gland adenocarcinoma, and to study the relation between the E6/E7 gene and squamous differentiation. Plasmid pBR322 containing HPV-16 was transfected into cultured SGT cell line using lipofectin method. Hygromycin was used as a selection marker. The presence of HPV E6/E7, transglutaminase 1, and involucrin mRNAs and protein in E6/E7 gene transfected cells was investigated by RT-PCR and immunoslot blot method. The apoptosis index was analysed by flow cytometry. The growth rate of E6/E7 gene transfected cells was reduced. E6/E7 transfected SGT cells increased apoptosis index. Involucrin and TGase I mRNAs by the squamous cell differentiation was most conspicuous in the E6/E7 gene transfected cell compared with non transfected cells. Squamous cell differentiation demonstrated in the transfectedSGT cell line, which expressed E6/E7 fusion gene mRNA.E6/E7 gene transfected cells showed squamous cell differentiation, expressing involucrin and TGase 1 protein by immunoslot blotting. The transfected SGT cell which expressed E6/E7 gene mRNA showed the squamous cell differentiation particularly clearly, and apoptosis was also demonstrated. It suggested that E6/E7 gene transfection into human salivary gland adenocarcinoma cells might induce clear squamous cell differentiation and contribute to study the pathogenesis of human salivary gland adenocarcinoma.

Urokinase-type plasminogen activator (uPA) and plasminogen activator type 1 (PAI-1) inhibitor contribute to the invasiveness of many carcinomas. It will be helpful to study clinical behavior of patients with malignant tumor by analysis of their expression. Expression of uPA and PAI-1 in human salivary gland tumors has been rarely reported in vitro. The purpose of this study were to investigate the protein expression of uPA and PAI-1 in SGT cell line compared to oral SCC and HeLa cell lines and to study migration and adhesion assay. All the cell lines were cultured under DMEM with 10% FBS at at 37oC in a 5% CO2 incubator. We studied a possible association between cytosolic uPA and PA-1 concentrations in SGT cell line compared to any other cell lines through cell migration and adhesion assay, and enzyme-linked immunoassay(ELISA). In migration assay SGT cell line was about 2 .5-4 folds higher than another cell lines. In adhesion assay SGT cell line was about 1.1-2 folds higher than another cell lines. uPA cytosolic concentrations of SGT cell line was about 3-10 folds, while PAI-1 was about 2.5-10 folds. Oral SCC cell lines were the lowest value. Both uPA and PAI-1 concentrations were correlated with migration and adhesion assay. High cytosolic concentrations of uPA and PAI-1 was correlated with migration and adhesion assay. It suggested that these markers might be specific marker for SGT cell line and would be contributed to treatment and prognosis of human salivary gland adenocarcinoma

Oral squamous cell carcinoma (SCC) is one of the leading causes of cancer mortality worldwide. It is generally thought that adjuvant chemotherapy provides modest prolongation of survival in various carcinoma. Docetaxel (Taxotere, TXT) play a significant role in the treatment of various solid tumors of epithelial origin. CsA (immunosuppressive drug) was widely used as adjunct for the treatment of cancer. Thus, it is important to pursue the apoptosis of IHOK and oral SCC induced by TXT combined with CsA related to the pathogenesis of oral SCC. But TXT combined CsA effect on IHOK and oral SCC remains unclear. After cultured IHOK and HN 22 oral squamous cell carcinoma cell line treated by 10 nM TXT and 1 μM, and caspase inhobitor, respectively, apoptosis index, cytochrome c and caspase-3 -8, -9 mRNA expression by RT-PCR, and procaspase-3 protein amount by immunoslot blotting was prepared. The purpose of this study were to examine the TXT-induced apoptosis pathway via caspase activation by CsA enhancement, and to apply these results to an effective therapeutic treatment plan for oral SCC by TXT combined CsA . 10 nM TXT showed about 60%, 55% celluar apoptosis of IHOK and HN 22, cell line, respectively, while CsA alone did not induce apoptosis in IHOK and HN 22 cell line. 1 μM CsA combined with 10 nM TXT increased apoptosis in IHOK and HN 22 cell line through caspase-3 and cytochrome c mRNA expression, while could not effect on caspase-8 and -9. Caspase inhibitor suppressed apoptosis of IHOK and HN 22 cell line induced by a combination of 1 μM CsA and 10 nM TXT. Immnoslot blotting showed procaspase-3 activation by a combination 1 μM CsA and 10 nM TXT, while caspase inhibitor inhibited activation. It suggested that a combination of CsA and TXT might induce increased apoptosis of IHOK and HN 22 oral squamous cell carcinoma cell line through caspase-3 activation. This treatment with a combination of TXT and CsA may be an effective therapeutic strategy for oral squamous cell carcinoma