Hwang Jin-Ming, Kao Shao-Hsuan, Hsieh Yi-Hsien, Li Kuen-Lin, Wang Pei-Hsi, Hsu Li-Song, Liu Jer-Yuh
Department of Applied Chemistry, School of Medicine, College of Medicine, Chung Shan Medical University, Taichung 402, Taiwan.
Mol Cell Biochem. 2009 Jul;327(1-2):135-44. doi: 10.1007/s11010-009-0051-3. Epub 2009 Feb 18.
Anion exchanger (AE) 2, belonging to the chloride-bicarbonate transporter family, has been reported to involve cell survival for hepatocellular carcinoma (HCC) cells. Our previous findings showed that AE2 gene was highly expressed in a poorly differentiated HCC cell line, HA22T/VGH. Additionally, treatment with 4,4'-diisothiocyanatostilbene-2,20-disulfonic acid (DIDS), an AE-specific inhibitor, significantly inhibited cell proliferation and induced cell apoptosis for the HA22T/VGH. To further investigate the biological functions of AE2 in human HCC, suppression of AE2 expression by the antisense oligonucleotide-AE2 (AS-AE2) was performed, and the cell viability, cell cycle regulation, and cell apoptosis for HCC cell lines were monitored. The results showed that AS-AE2 treatment could efficiently suppress the mRNA expression of AE2 for various differentiated HCC cells, including HA22T/VGH, SK-Hep-1, PLC/PRF/5, Hep3B, and HepG2. Moreover, AS-AE2 treatment significantly reduced cell viability, arrested cell cycle at sub-G1 phase, and induced cell apoptosis for the poorly differentiated HA22T/VGH, but not for other moderately or well-differentiated HCC cell lines. The findings indicated that AE2 may play an important role in the progression of HCC cells, and provide a new strategy for the development of therapeutic treatment against human HCC.
阴离子交换蛋白（AE）2属于氯-碳酸氢根转运体家族，据报道其与肝细胞癌（HCC）细胞的存活有关。我们之前的研究结果表明，AE2基因在低分化的肝癌细胞系HA22T/VGH中高表达。此外，用AE特异性抑制剂4,4'-二异硫氰酸芪-2,2'-二磺酸（DIDS）处理，可显著抑制HA22T/VGH细胞的增殖并诱导其凋亡。为了进一步研究AE2在人类肝癌中的生物学功能，我们采用反义寡核苷酸-AE2（AS-AE2）抑制AE2的表达，并监测肝癌细胞系的细胞活力、细胞周期调控和细胞凋亡情况。结果表明，AS-AE2处理可有效抑制包括HA22T/VGH、SK-Hep-1、PLC/PRF/5、Hep3B和HepG2在内的各种分化程度的肝癌细胞中AE2的mRNA表达。此外，AS-AE2处理显著降低了低分化的HA22T/VGH细胞的活力，使细胞周期停滞在亚G1期并诱导其凋亡，但对其他中度或高分化的肝癌细胞系没有影响。这些发现表明，AE2可能在肝癌细胞的进展中起重要作用，并为开发针对人类肝癌的治疗方法提供了新策略。
