反譯寡核甘酸AE2對於人類肝細胞癌細胞存活率之影響

最近利用了mRNA差別顯示技術 (differential display reverse transcriptase polymerase chain reaction；DDRT-PCR) 在不同惡化程度的肝癌細胞內鑒定和比較不同基因的表現。我們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。為了了解AE2在人類肝細胞癌內的生物功能和作用機制，我們進行反譯寡核甘酸AE2 (antisense oligonucleotide AE2) 實驗，結果發現在HA22T/VGH、SK-Hep-1、PLC/PRF/5、Hep3B 和HepG2的肝癌細胞株中，只有HA22T/VGH細胞的生長速率有明顯的下降，而且有劑量和時間依賴現象。接著利用DAPI (4-6-di-amidino-2- phenylindole) staining、DNA fragmentation和flow cytometry方法分析確認，結果顯示反譯寡核甘酸AE2會造成HA22T/VGH細胞凋亡。以上結果指示AE2在人類肝細胞癌惡化過程中可能扮演重要角色，同時也將提供新的基因治療策略。

關鍵字

肝癌、細胞凋亡、陰離子交換通道蛋白2、反譯寡核甘酸AE2

並列摘要

Recently, we identified and characterized differentially expressed genes in the different malignant human hepatocellular carcinoma (HCC) cell lines by using (Differential Display Reverse Transcriptase Polymerase Chain Reaction; DDRT-PCR). We fund that an anion exchanger 2 (AE2) gene was highly expressed in HA22T/VGH cells. To understanding the biological function and gene regulation of AE2 in human HCC, the antisense oligonucleotide AE2 assay was performed. The results showed that cell proliferation was decreased in a dose- and time-dependent manner only in HA22T/VGH cells, but not in other cell lines (K-Hep1, PLC/PRF/5, Hep3B and HepG2). Furthermore, by using DAPI (4-6-Di-Amidino-2-PhenylIndole) staining, DNA fragmentation and flow cytometric analysis, these results confirmed that antisense oligonucleotide AE2 induced cell apoptosis in HA22T/VGH cells . These findings indicated that AE2 may play an important role in the progression of human HCC, and may provide new strategy for gene therapy.

並列關鍵字

Hepatocellular carcinoma、Apoptosis、anion exchanger 2、antisenseoligonucleotide AE2

參考文獻

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反譯寡核甘酸AE2對於人類肝細胞癌細胞存活率之影響

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