Upregulation of ferritin H(L)chain gene in cultured human HepG2 cells induced by arsenical trioxide / 解放军医学杂志

ABSTRACT

Objective In order to understand the differentially expressed genes and explore the effects on mechanism of gene expression induced by arsenic trioxide. Methods The mRNA was isolated from human HepG2 cells treated with arsenic trioxide( 5?mol/L ) and DMSO, respectively, then cDNA was synthesized. After restriction enzyme Rsa Ⅰ digestion, small sizes cDNA were obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. After tester cDNA was hybridized with driver cDNA twice and underwent nested polymerase chain reaction (PCR) twice, the DNA fragment was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after colony PCR. Results The forward subtracted cDNA library from HepG2 cell line induced by arsenic trioxide was successfully constructed. The sequencing analysis showed that there were eight clones contained ferritin H(L) chain in the library. Conclusion Arsenic trioxide can induce the up expression of ferritin H(L) chain protein in HepG2 cells, indicated that the ferritin H(L) chain may play certain role in the mechanism of anti-arsenical cytotoxicity in liver.