生長因數對人類乳突狀第十六型病毒DNA轉型之人類鼻腔上皮細胞中病毒基因表現，細胞生長及DNA合成調控之研究

Abstract

為探討人類乳突狀第十六型病毒(Human Papillooma-virus Type16, HPV-16)的致癌特性，我們將完整的HPV-16 DNA輿質體(plasmid) pdMMT之DNA重組，並將重組後的DNA轉入正常人類鼻腔上皮細胞，結果這細胞成功地被轉型為不死的細胞株，命名為NW-1。由生長需求實驗中顯示NW-1細胞不死的現像是牛腦下垂體萃取(Bovine Pituitary Extract, BPE)表皮生長因數(Epidermal Growth Factor，EGF)或胰島素(Insulin，Ins)與HPV-16 DNA相互作用的結果。 當細胞培養在生長較佳的狀況下，Northern Hybridization的實驗結果發現細胞有較多的HPV-16所有基因轉錄?物。當細胞以BPE,EGF和Ins或EGF及Ins 處理20小時後，RNA slot blot Hybridization的實驗結果證明HVP-16所有基因的表現也會增加。HPV-16之E6及E7基因的表現利用RNA Polymerase Chain Reaction (RNA-PCR)偵測，結果發現細胞培養在不同的狀況下均有三種放大產物出現，其大小各?791,608及491Base Pairs，這些RNA-PCR產物量之多寡輿細胞生長之快慢或DNA合成並沒有直接的正相關。

To determine the oncogenic properties of Human Papillomavirus Type 16(HPV-16), we have been successful in transfecting normal human nasal epithelial cells with recombinant DNA containing complete HPV—16 genomic DNA. The results of growth requirement indicate that the immortalization of NW—1 cells likely is due to the interaction between the growth factors and HPV-16 DNA . Greater amount of total viral transcripts was found in a culture condition in which NW—1 cells grow relatively well and induction of HPV—16 total transcripts occurres 20hr after treating the cells with 5ng/ml Epidermal Growth Factor and 10 μg/ml Insulin with or without 15 μg/ml bovine pituitary extract. Additionally, RNA polymerase chain reaction (PCR) was used to analyze the transcripts of E6 and E7 genes of HPV-16 in NW—1 cells cultured in MCDB—151 supplemented with different growth additive. Three expected amplification products of 791,608 and 491 base pairs in length were identified in NW—1 cells cultured differently. Contrary to what have been thought, however, DNA synthesis and cell growth of NW—1 showed no direct correlation with quantitative differences of PCR products resulting from different culture conditions.