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

黎琨林(Kuen-Lin Li)

指導教授 ： 劉哲育

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2005年)

https://doi.org/10.6834/CSMU.2005.00042

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。

關鍵字

陰離子交換通道蛋白二 ； mRNA差別顯示技術

並列摘要

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.

並列關鍵字

Anion exchanger 2 ； Differential Display Reverse Transcriptase Polymerase

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