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Received for publication, April 24, 2003Published, JBC Papers in Press, April 28, 2003, DOI 10.1074/jbc.M304307200
Tommi J. Ahonen‡§¶, Jianwu Xie‡¶, Matthew J. LeBaron‡, Jianqiong Zhu‡, Martti Nurmi , Kalle Alanen**, Hallgeir Rui‡, and Marja T. Nevalainen‡ ‡‡
Inhibition of Transcription Factor Stat5 Induces Cell Death of Human Prostate
Cancer Cells*
班級 :生科四甲學生 :梁育銘
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男性泌尿生殖系統構造圖男性泌尿生殖系統構造圖
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症狀症狀
早期的攝護腺癌是沒有症狀的。當攝護腺早期的攝護腺癌是沒有症狀的。當攝護腺癌侵犯到尿道時癌侵犯到尿道時 , , 病患會 有頻尿、夜尿、病患會 有頻尿、夜尿、尿急、尿流細小、尿道疼痛和血尿等症狀尿急、尿流細小、尿道疼痛和血尿等症狀 , , 和良性 攝護腺肥大的症狀類似。當攝護腺和良性 攝護腺肥大的症狀類似。當攝護腺癌轉移到骨骼癌轉移到骨骼 , , 如腰椎骨時如腰椎骨時 , , 就 會造成就 會造成腰背疼痛以神經痛的症狀。腰背疼痛以神經痛的症狀。
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IntroductionIntroduction
攝護腺癌常見於攝護腺癌常見於 50 50 歲以上的男性。在美國僅次於肺癌 及大腸 歲以上的男性。在美國僅次於肺癌 及大腸 癌,是男性常見的癌病,美國 的罹患率是每十萬癌,是男性常見的癌病,美國 的罹患率是每十萬人口有七千人(或 者說每六個美國男性就有一人,人口有七千人(或 者說每六個美國男性就有一人,在一 生當中會罹患此病),以 在一 生當中會罹患此病),以 1995 1995 年為例, 在年為例, 在所有該年新檢測出的癌症之中,攝 護腺癌就佔了 所有該年新檢測出的癌症之中,攝 護腺癌就佔了 36%36% 。每年有 。每年有 240,000240,000 個新的病例被檢出,並造個新的病例被檢出,並造成超過四萬人的死亡。成超過四萬人的死亡。
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AbstractAbstract
從從 4040 個病患身上所取得的攝護腺個病患身上所取得的攝護腺腫瘤組織切片腫瘤組織切片 ,, 在其樣本中我們發在其樣本中我們發現大約現大約 2626 個人個人 (65%),(65%), 它的它的 Stat5Stat5有異常活化的現象有異常活化的現象 ,, 因此作者大膽因此作者大膽假設異常活化的假設異常活化的 Stat5Stat5 和攝護腺腫和攝護腺腫瘤有一定的關係瘤有一定的關係 ,, 所以它提出阻斷所以它提出阻斷Stat5Stat5 的功能可能是一種治療攝護的功能可能是一種治療攝護腺腫瘤的一個新方法腺腫瘤的一個新方法。。
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STATs 蛋白質簡介 :STATs 蛋白質的發現 ,最早是由於干擾素的研究 ,由於 INF 刺激細胞後 ,會產生一群特定基因的表現 ,而發現這些基因的表現 ,是需要 STATs 蛋白質的參與作用 ,STATs蛋白質它擁有兩種生物功能 :
1. 將訊息由細胞表面傳到細胞核內2.STATs 蛋白質也參與其基因表現調控
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STATs Family:
STATs Family 的成員有七個 :
包括 STAT1. STAT2….. 到 STAT6, 除了 STAT2 .STAT6較大 ,大約是 850 個 amino acids, 其它大約是 700-800 個 amino acids , 然後 STAT5 它有兩個同源性的基因產物分別是
STAT5a- 94-kDa (Tyr-694)
STAT5b- 92-kDa (Tyr-699) JAK2
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STAT3 Signaling Pathway
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名詞介紹Ad- adenovirusAd- adenovirus
為一個 DNA 腫瘤病毒 ,可做為真核細胞基因表現之研究
DNStat5- dominant-negative Stat5DNStat5- dominant-negative Stat5 顯性 -抑制型的 Stat5, 為 Stat5 的一個 mutant, 喪失其活化功
能AdDNStat5-Adenoviral gene delivery of a dominanAdDNStat5-Adenoviral gene delivery of a dominant-negative Stat5t-negative Stat5
STAT-Signal transducer and activator of transcription
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Materials
1.CWR22Rv 2.LNCap
3.Pc-3 cells
從 40個病人身上所取得的攝護腺腫瘤組識切片
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Anti-phospho-TyrStat5 Anti-pan-Stat5
Human prostate cancer cells
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Normal human prostate epithelium
Anti-phospho-TyrStat5
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A dose-dependent effect on cell viability of adenoviral gene delivery of a dominant-negative Stat5 mutant (AdDNStat5)
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Morphology of cell death induced by AdDNStat5 in CWR22Rv and LnCap cells
( 72h , 8 m.o.i) ( 96h , 32 m.o.i)
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Dose-dependent expression of WTStat5 or DNStat5 in CWR22Rv and LnCap cells
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DNA fragmentation analyzed by nucleosomal ELISA after exposure to AdDNStat5, AdWTStat5,AdlacZ, AdCtrl, or mock infection in CWR22Rv and LNCap
0.3
2.1
0.4
2.4
( 48h , 8 m.o.i) ( 72h , 32m.o.i)
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用 Flow Cytometry 測量細胞 DNA 含量和細胞的大小 , 來檢測凋亡細胞
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DNStat5-induced cell apoptosis involved caspase-3 activation
( 36h , 8 m.o.i) ( 48h , 32 m.o.i)
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DNStat5- induced caspase-3 activation is associated with activation of caspase-9.
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結論用原位偵測法去測定磷酸化和細胞核的 Stat5, 在大部份我們檢測的攝護腺腫瘤細胞發現它的 Stat5 是活化的 ,所以我們提出如果抑制 Stat5 的活化可以是一種新的基因療法 ,它可以廣泛應用在治療攝護腺腫瘤細胞上 ,我們觀察到 CWR22Rv 和 LNCap 這兩個腫瘤細胞中 ,阻斷 Stat5 的活化可以引發一連串的反應 ,如 caspase-3caspase-9和的活化最後誘導整過攝護腺腫瘤細胞 apoptosis,來達到治療腫瘤的效果 .