陰離子交換通道抑制劑(DIDS)對於肝癌細胞株之生長率影響

中山醫學大學機構典藏 CSMUIR > 醫學院 > 生化微生物免疫研究所 > 博碩士論文 > Item 310902500/617

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题名:	陰離子交換通道抑制劑(DIDS)對於肝癌細胞株之生長率影響 Effect of anion transport inhibitor (DIDS ) on cell proliferation of human hepatoma cells
作者:	黎琨林 Kuen-Lin Li
贡献者:	中山醫學大學：生化暨生物科技研究所;劉哲育
关键词:	陰離子交換通道蛋白二;mRNA差別顯示技術 Anion exchanger 2;Differential Display Reverse Transcriptase Polymerase
日期:	2005/07/20
上传时间:	2010-03-12T02:25:53Z (UTC)
摘要:	本實驗室最近就專研於肝細胞癌（Hepatocellular carcinoma；HCC）的研究，利用了mRNA差別顯示技術（Differential Display Reverse Transcriptase Polymerase Chain Reaction；DDRT-PCR) 的技術偵測不同惡化程度的肝癌細胞，發現到陰離子交換通道蛋白二 (Anion Exchanger 2；AE2) 基因在不同分化程度的細胞株內有不同的表現。陰離子交換通道蛋白二在生理學上的功能具有調節pH值、細胞體積、重碳酸鹽的運送和維持細胞內氯離子的濃度。本實驗從藥理學的方向研究 AE2在四株肝癌細胞株 (Sk-Hep-1、HA22T/VGH、HepG2及Hep3B)中的功能的表現。結果顯示當處理Anion transport inhibitor 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS)可以造成HA22T/VGH細胞的生長速率下降，其他細胞株沒有影響。利用DAPI (4-6-Di-Amidino-2-PhenylIndole) staining、DNA fragmentation和flow cytometry分析方法確認DIDS會造成HA22T/VGH細胞凋亡。同時DIDS也證實會抑制HA22T/VGH細胞內AE2的活性。所以我們認為DIDS造成HA22T/VGH細胞凋亡可能與抑制AE2活性有關。 Recently, we have investigated human cellular carcinoma (HCC) in our laboratory, and used DDRT-PCR to detect different liver cell lines. We fund that a AE2 gene was highly expressed in HA22T/VGH cells. AE2, a member of the Cl–/HCO3– exchanger family, contributes to the regulation of intracellular pH, intracellular Cl– concentration and is regulated by acid-base balance. In this study, we investigate the pharmacological function of AE2 in four hepatoma cell lines (HepG2、HA22T/VGH、SK-HEP-1 and Hep3B). Treatment with Anion transport inhibitor 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS), cell proliferation was decreased in a dose- and time-dependent manner only in HA22T/VGH cells, but not in other cell lines. Using DAPI (4-6-Di-Amidino-2-PhenylIndole) staining、DNA fragmentation and flow cytometric analysis, these results confirmed that DIDS may induced cell apoptosis in HA22T/VGH cells. DIDS was also demonstrated to inhibit the activity of AE2 in HA22T/VGH cells. These results indicated that the cell apoptosis induced by DIDS may be related with the decrease in AE2 activity in HA22T/VGH cells.
URI:	http://140.128.138.153:8080/handle/310902500/617
显示于类别:	[生化微生物免疫研究所] 博碩士論文

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