Nrf2在不同肝癌细胞株中的表达及定位

Expression and distribution of Nrf2 in several hepatocellular carcinoma cell lines

目的:观察红系衍生的核因子2相关因子2(Nrf2)在肝癌细胞株(HepG2、Hep3B、SMMC-7721)中的表达及分布情况。方法:应用流式细胞术测定Nrf2阳性细胞的比例,激光共聚焦观察Nrf2在3种细胞株中的定位情况,通过Westernblot测定Nrf2在不同细胞胞质及胞核中的表达差异。结果:流式细胞术测定Nrf2在HepG2、Hep3B和SMMC-77213种细胞中表达率分别为99.39%、99.94%和99.98%,激光共聚焦观察到Nrf2在3种肝癌细胞株胞质胞核中均有分布,但不同细胞的分布规律各不相同,Western blot结果显示:Nrf2在HepG2中以胞质分布为主,在Hep3B中胞质、胞核分布相当,在SMMC-7721中以胞核分布为主。结论:Nrf2在3种肝癌细胞株(HepG2、Hep3B、SMMC-7721)中均有表达,且在胞质胞核中均有分布,但在不同细胞中的分布规律各不相同,为下一步探讨肝癌细胞耐药机制的相关研究奠定了基础。

AIM： The aim of this study was to investigate the expression and the distribution of Nuclear factor-erythroid 2 p45-related factor 2（Nrf2） in several human hepatocellular cancer cell lines（HepG2,Hep3B,SMMC-7721）.METHODS： Flow cytometry was used to figure out the percentages of Nrf2-positive cells in these three cell lines.The localization of Nrf2 was estimated by the laser confocal microsopy and the expression levels of Nrf2 in hepatocellular cancer cell lines were detected by Western blot analysis.RESULTS： The percentages of Nrf2 positive cells in HepG2,Hep3B,and SMMC-7721 were 99.39%,99.94%,and 99.98% through the flow cytometry.The laser confocal microsopy showed that Nrf2 mainly localized in the cytoplasm of HepG2 cells,distributed evenly in the cytoplasm and nucleus of Hep3B cells,and mainly localized in the nucleus of SMMC-7721 cells.Western blot analysis confirmed the result by the laser confocal microsopy.CONCLUSION： The data on the expression and localization of Nrf2 will be helpful for the following research on the role of Nrf2 in the drug resistance of hepatocellular carcinoma to chemotherapy.