[Effect of triptolide on reversing hypermethylation of apc gene in Jurkat cells and its possible mechanisms].
This study was aimed to investigate the effect of traditional Chinese medicine, Triptolide (TPL) on reversing hypermethylation of antioncogene (apc gene) in acute lymphoblastic leukemia cell line Jurkat in vitro and to explore its mechanisms. The effects of TPL on cell growth, proliferation and cell cycle were detected by growth curve, MTT assay, colony formation test and flow cytometry, respectively. The effect of TPL on apc gene methylation of Jurkat cells was analyzed by nested methylation specific PCR; the expressions of apc gene, dnnt3a, dnmt3b mRNA were measured by RT-PCR; the protein expression of apc gene was detected by Western blot. The results showed that as compared with untreated control cells, the TPL of different concentrations could significantly inhibit growth and proliferation of Jurkat cells in dose-and time-dependent manners with IC₅₀ 19.7 ng/ml at 48 hours. All cytosines in CpG dinucleotides in untreated Jurkat cells had no changed, while all cytosines in Jurkat cells treated with TPL had been converted to thymidine suggesting the methylation of apc gene in Jurkat cells. The TPL could reverse hypermethylation of apc gene and induced the mRNA and protein expression of apc gene in dose-dependent manner. It is concluded that the small dose of TPL can obviously suppress the proliferation of Jurkat cells, activate and up-regulate the expression of apc gene through demethylation of apc gene resulting from DNMT and/or direct action, thereby inhibit the proliferation rate of Jurkat cells.
Wu
Xue-Mei
XM
Department of Hematology, Fuzhou Children's Hospital, Fuzhou 350004, Fujian Province, China.
Shen
Jian-Zhen
JZ
Shen
Song-Fei
SF
Fan
Li-Ping
LP
D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects*; D000090205:DNA Methyltransferase 3A; D004224:Diterpenes / Q000494:pharmacology*; D004852:Epoxy Compounds / Q000494:pharmacology; D015973:Gene Expression Regulation, Leukemic; D017491:Genes, APC / Q000187:drug effects*; D006801:Humans; D019169:Jurkat Cells; D010616:Phenanthrenes / Q000494:pharmacology*; D000097584:DNA Methyltransferase 3B
D018906:Antineoplastic Agents, Alkylating; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D004224:Diterpenes; D004852:Epoxy Compounds; D010616:Phenanthrenes; D000097584:DNA Methyltransferase 3B; C000717967:DNMT3A protein, human; C001899:triptolide