본 연구의 목적은 계획된 행위이론에 근거한 HPV 예방교육이 대학생의 HPV 백신지식, 자궁경 부암 지식, HPV 백신접종에 대한 태도, HPV 백신접종에 대한 주관적 규범, HPV백신접종에 대한 지각된 행위통제, HPV 백신접종에 대한 의도, HPV 예방백신 접종행위에 미치는 효과를 확인하기 위한 유사실험 연구이다. 대상자는 실험군 32명, 대조군 34명으로 임의표출 하였다. 수집된 자료는 기술통계, t-test, χ 2-test, Fisher’s exact test와 repeated Measures ANOVA로 분석하였다. 연구결과 HPV 예방교육에 참여 한 실험군은 참여하지 않은 대조군과 HPV 백신지식(t=5.66, p<.001), 자궁경부암 지식(t=4.13, p<.001), 태도(t=2.24, p=.032), 주관적 규범(t=2.83, p=.008), 지각된 행위통제(t=2.65, p=.013), 의도(t=3.91, p<.001)에 유의한 차이가 있었다. HPV 예방교육 중재 4주 후 HPV 백신접종 의도는 집단과 시간의 경과 간의 교호작용에 유의한 차이가 있었다(F=6.95, p=.002). 따라서 HPV 예방교육은 대학생에게 실제 적용 할 수 있는 교육프로그램임을 확인하였다.

Cellular microenvironment is an essential issue for regulating epithelial characteristics through the alteration of intricate signaling pathways and intercellular communications in different cell types. Thus, microenvironment influences tumor initiation, progression, and metastasis. This study aimed to investigate the relationship between microenvironment and epithelial property in HPV16 E6/E7-immortalized human oral keratinocytes (IHOKs). To investigate characteristics of IHOK cultured in different media, two media were used, which included keratinocyte growth media (KGM), F-medium composed of 3:1 ratio of DMEM and F-12 (P media) supplemented with 10% FBS and 1% penicillin/streptomycin. Proliferative property and invasive and migratory activity were observed. As results, proliferating activities of IHOK in different culture condition were changed. Likewise, migratory and invasive activities were also different depending on media types. These results suggest that cellular microenvironment can affect modification of biological properties of epithelial cells.

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.

Human papillomavirus (HPV) has been classified as one of the causing factors of head and neck squamous cell carcinoma (HNSCC). However, little is known about HPV-related carcinogenesis in HNSCC. The purpose o f this s tudy i s to characterize immortalized human oral keratinocyte (IHOK) transfected by HPV16 E6/E7, IHOK/hcdk4 (IHOK transfected by pLXRN-hcdk4) and IHOK/hcdk4/hTERT (IHOK transfected by pLPC-hTERT-hcdk4) to reconstitute HNSCC in vitro. Conclusively, we established a new immortalized cell lines, IHOK/hcdk4 and IHOK/hcdk4/hTERT, to understand multistep carcinogenic process of oncogenic HPV16 E6/E7 in HNSCC.

Studies to evaluate distribution of markers in normal keratinocyte and their immortalized keratinocyte are appropriate to evaluate the normal and preneoplastic lesion of oral cancers as biochemical and cytochemical changes associate with tumorigenesis being not completely understood. Complementary DNA microarray containing 6000 sequence -verified cDNA elements was used to systematically characterize the variation in gene expression patterns of NHOK cells vs. immortalized keratinocyte by HPV16 E6-E7(IHOK). Examination of gene expression that is 85 clones cDNAs exhibits greater than 2 fold overexpression in NHOK probes relative to IHOK probe, 147 cDNAs reveal greater 2 fold overexpression in IHOK relative to NHOK probe.The high similarity in gene expression (96.5%) between IHOK and NHOK cells suggests that only an additional 232/6720 (3.5%) of the genome is differentially gene activated during HPV16 immoratlized keratinocyte growth and differentiation. Examination of gene expression that differs between NHOK and IHOK cellsapprear to be related to : cell adhesion & recognition, cell cycle regulator, apoptosis, transciption factors, growth factors and therir receptors, cytoskeletal and extracellular matrix proteins, signal transduction modulators and effectors, and miscellaneous. The gene expression of cell recognition factor such as endothelin 1, collagen IV, fibronectin, and SPR1 in IHOK were upregulated. Distinct or duplicated cDNA clones representing the same gene were typically clustered in adjacent rows in the clustered gene map. Therefore the differentially expressed and identified genes should be informative in studying oral epithelial cell carcinogenesis and such studies should foster the research of molecular markers allowing to assess the phenotypeof malignant epithelial tumor.

Epithelial-mesenchymal interaction is well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular micro-environment which provide a epithelial-mesenchymal interaction. The aims of this study were to develop and evaluate the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted human normal oral kertinocyte(NHOK) and immortalized human oral keratinocytes(IHOK) by histological and immunohistochemical analysis. The results were as follows; 1. Best condition of three dimensionally reconstituted IHOK & HaCaT cells are 14 days air-exposure cultivation, 3 days of submerged state, and dermal equivalent consisting type I collagen and IGF cells. 2. In comparison to IHOK, there was better preservation of the overall epidermal strucutures in oral cracioma cells (HN30) & HaCaT cells by organotypic cultures. But orgnaotypic co-culture of the normal keratinocyte showed the thinnest epithelial layer formation. 3. PCNA was detected primarily in the basal layer of normal mucosa and NHOK, whereas was shown throught the epithellium except surface layer of IHOK cells, and it's expression was similar to that of CIS of biopsied patient's tissue 4. Involucrin is expressed in the upper layer of oral mucosa and NHOK raft, but staining for involucrin was induced in the IHOK rafts indicating differentiation is incomplete, and the staining pattern in the IHOK raft was not uniform. 5. Normal oral keratinocyte raft showed weak immunostaining for p53, and p53 expression of IHOK raft increased rahter than in NHOK. In organotypic cultures of normal cells and IHOK, p53 expression was restricted to the proliferative part of epithelium. This is consistent with expression pattern in biopsy specimens of the normal and CIS tissue. 6. In artifically reconstructed NHOK, the pattern of keratin staining showed both similarity and differences from that of intact normal mucosa. An obvious difference was increased expression of CK10 & CK19, and decreased expression of CK6 in a reconstructed NHOK rather than in normal mucosa, and similar expression was in CK4 and CK16. 7. CK19 & CK16 were strongly positive in HPV immortlaized keratinocyte rafts rahter than in NHOK, an indicator of premalignant or malignant changes, while CK10 & CK6 were decreased, and organotypic cultures of IHOK express was similar keratin expression as epithelial dysplasia or CIS tissue. These results suggest that three-dimensional organotypic co-culture of normal oral & immortalized keratinocytes with dermal equivalent consisting type I collagen and fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. Thus this organotypic system can be used for studing mechanism of epitehlial-mesenchymal interaction in particular regulating epidermal diffenentiation and morphogenes

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.