제16형 HPV E6/E7 유전자의 변형 감염으로 배양된 SGT 세포주에서 유도된 편평세포 분화

서지정보

ㆍ발행기관 : 대한구강악안면병리학회 ㆍ수록지정보 : 대한구강악안면병리학회지 / 34권 / 3호
ㆍ저자명 : 이종현, 박경주, 이종헌

목차

I. INTRODUCTION
II. MATERIALS AND METHODS
1. Cell Culture
2. HPV DNA
3. Transfection of HPV DNA into Cultured SGTCell Line
4. Growth Curve and Apoptosis Index
5. Western blot analysis
6. Detection of HPV mRNA by RT-PCR
III. RESULTS
1. Growth Curve and Apoptosis Index
2. Detection of HPV E6 and E7 mRNA
3. mRNA Determinationof Involucrin andTransglutaminase 1
4. Immunoslot blot Analysis of Involucrin andTransglutaminase 1
V. DISCUSSION
VI. REFERENCES

영어 초록

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.

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