Zhao Hua, Tang Jiayong, Xu Jingyang, Cao Lei, Jia Gang, Long Dingbiao, Liu Guangmang, Chen Xiaoling, Wang Kangning
Animal Nutrition Institute, Sichuan Agricultural University, No 211 Huimin Road, Wenjiang, Chengdu, Sichuan, 611130, China,
Biol Trace Elem Res. 2015 Oct;167(2):236-41. doi: 10.1007/s12011-015-0323-6. Epub 2015 Apr 7.
The purpose of this study was to compare messenger RNA (mRNA) expression of selenoprotein genes between hepatoma HepG2 and normal hepatocytes LO2 cell lines. Liver HepG2 and LO2 cells were cultured in 12-well plates under the same condition until cells grew to complete confluence, and then cells were harvested for total RNA and protein extraction. The qPCRs were performed to compare gene expression of 14 selenoprotein genes and 5 cancer signaling-related genes. Enzyme activities were also assayed. The results showed that human hepatoma HepG2 cells grew faster than normal hepatocytes LO2 cells. Among the genes investigated, 10 selenoprotein genes (Gpx1, Gpx3, Gpx4, Selx, Sepp, Sepw1, Sepn1, Selt, Seli, Selh) and 3 cancer signaling-related genes (Bcl-2A, caspase-3, and P38) were upregulated (P < 0.05), while Selo and Bcl-2B were downregulated (P < 0.05) in hepatoma HepG2 cells compared to LO2 cells. Significant correlations were found between selenoprotein genes and the cancer signaling-related genes Caspase3, P53, Bc1-2A, and Bc1-2B. Our results revealed that selenoprotein genes were aberrantly expressed in hepatoma HepG2 cells compared to normal liver LO2 cells, which indicated that those selenoprotein genes may play important roles in the occurrence and development of liver carcinogenesis.
本研究的目的是比较肝癌HepG2细胞系和正常肝细胞LO2细胞系中硒蛋白基因的信使核糖核酸(mRNA)表达。将肝癌HepG2细胞和LO2细胞在相同条件下培养于12孔板中,直至细胞完全汇合,然后收获细胞用于总RNA和蛋白质提取。进行定量聚合酶链反应(qPCR)以比较14个硒蛋白基因和5个癌症信号相关基因的基因表达。同时也测定了酶活性。结果显示,人肝癌HepG2细胞比正常肝细胞LO2细胞生长更快。在所研究的基因中,与LO2细胞相比,肝癌HepG2细胞中有10个硒蛋白基因(Gpx1、Gpx3、Gpx4、Selx、Sepp、Sepw1、Sepn1、Selt、Seli、Selh)和3个癌症信号相关基因(Bcl-2A、半胱天冬酶-3和P38)上调(P < 0.05),而Selo和Bcl-2B下调(P < 0.05)。在硒蛋白基因与癌症信号相关基因半胱天冬酶3、P53、Bc1-2A和Bc1-2B之间发现了显著相关性。我们的结果表明,与正常肝脏LO2细胞相比,硒蛋白基因在肝癌HepG2细胞中异常表达,这表明这些硒蛋白基因可能在肝癌发生和发展中起重要作用。