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研究生:林重宏
研究生(外文):Lin, chung-hong
論文名稱:以反向寡核酸來研究抑癌基因p53在卵巢癌發生之角色
論文名稱(外文):The role of p53 in ovarian carcinogenesis-study by antisense p53
指導教授:陳燕惠陳燕惠引用關係
指導教授(外文):Chen,Yen-Hui
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:藥學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:1996
畢業學年度:84
語文別:中文
論文頁數:61
中文關鍵詞:卵巢癌反向寡核醣核酸p53 蛋白質
外文關鍵詞:ovarian cancerantisensep53
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p53 基因的變異或過度表現, 被發現與卵巢癌及其它癌症組織的發展及惡
化有關. 雖然p53 基因變異在卵巢癌組織常見, 但是對於它在卵巢癌所扮
演的真正角色, 卻不是很清楚, 為了更進一步瞭解p53 基因所參與的生長
調節角色與機制, 以p53 反向寡核醣核酸加入卵巢癌細胞來觀察 p53 蛋
白質, 細胞週期與其它相關因子的變化.PA-1 細胞株為一種具有過度表現
p53 蛋白質的卵巢癌細胞株, 將p53 反向寡核醣核酸加入PA-1細胞株, 並
觀察p53 蛋白質被抑制的程度. 結果顯示,lipofectin 加入與否, 皆能有
效進入PA-1 細胞株.當PA-1 細胞株加入50uM 的p53 反向寡核醣核酸, 可
以抑制p53 蛋白質的表現達百分之八十以上. 利用一系列有關細胞週期及
細胞計劃性死亡基因產物的抗體, 來探討p53 蛋白質被抑制後, 相關基因
產物的變化. 結果發現, PA-1 細胞中加入p53 反向寡核醣核酸50uM,
p21 蛋白質的表現下降了百分之三十, 而 CPP32及Ich-1l 蛋白質表現量
則上升. 此結果顯示,p53 蛋白質與 CPP32 , p21, 及Ich-1l基因產物表
現之調控有關.
Mutation and overexpression of p53 gene have been associated
with the development and progression of ovarian and other
cancers. In order to evaluate the involvement of p53 in
regulating proliferation and its mechanism of action in ovarian
cancer,we studied the association among expression of p53 gene
,cell cycle and related factors in ovarian cancer cells treated
with p53-modified antisense oligonucleotides. An ovarian cancer
cell line, PA-1, with overexpression of p53 was used in our
experiments. An 18-mer p53 antisense oligodeoxynucleotide(ODN)
covering the complementary strand of the initiating codon of
p53 was introduced into PA-1. We examined the efficiency of the
antisense ODN across PA-1. Results showed that the antisense
oligomers can be transported into PA-1 efficiently after
incubation for 24 hours with or without lipofectin, while the
antisense oligomers are poorly transported into the other cell
lines such as C33A.Inhibition of p53 protein expression by
antisense ODN was detected by western. Phosphothioated p53
antisense oligomers inhibit protein synthesis upto 80% in PA-1
cells when treated with 50uM antisense oligomers. A series of
antibodies against cell cycle and apoptosis associated gene
products were employed in western blot to study the downstream
effects of p53. Results showed that p21 protein synthesis was
30% inhibited when PA-1 cells were treated with 50uM p53 anti-
sense ODN for 24 hours. In contrast, we found that CPP32, Ich-1
l was enhanced when PA-1 cells were treated with 50uM antisense
ODN for 24 hours and 48 hours. The results suggest that
synthesis of p21, CPP32 and Ich-1l gene products is modulated
by p53. Treatment with p53 antisense ODN reverse the protein
synthesis of p21, CPP32 and Ich-1l.

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