The Effect Of 5-aza-dC-2’-deoxycytidine On Bovine Fetal Fibroblasts And Human Lung Cancer A549 Associated Mechanism

5-aza-dC-2-deoxycitidine (5-aza-dC) is used extensively as a demethylating agent, which effect genes expression by lowing the levels of methylation in cells. 5-aza-dC can significantly improve the efficiency of somatic cell nuclear transfer, clinical trial also showed that 5-aza-dC has therapeutic effect on cancer. However, only a few studies have been published regarding the effects of 5-aza-dC on cells and its mechanism. In this study, bovine fetal fibroblast cells and human lung cancer A549 cells were treated with 5-aza-dC respectively, then cell proliferation, cell cycle, cell apoptosis, reactive oxygen species and the mRNA expression of apoptosis related genes、pluripotent gene and cancer related gene were detected1. The effects of 5-aza-dC on bovine fibroblast cellsBovine fetal fibroblasts were treated with 0-10 mM 5-aza-dCfor 24 h and 48 h, respectively, results showed that compared to the untreated group,0-10 μM 5-aza-dC treatment cell viability, but has no significantly difference (p>0.05); while 5-aza-dC (1-10 mM) treatment decreased cell viability in a dose-and time-dependent manner (p<0.05). Flow cytometric analysis showed that 5-aza-dC increased cell apoptosis in a dose-dependent manner (p<0.05), cells were arrested in G0/G1 stage (p<0.05), and the concerntration of reactive oxygen species in cell raised rapidly after 5-aza-dC exposure. After treated with 10 mM 5-aza-dC for 48h, the mRNA expression of relative gene was measured by real-time PCR, results showed that 5-aza-dC treatment increased the expression of BAD, BAX, CYCS, CASPASE-3, CASPASE-9,while decresed the expression of BCL-2 in concentration-dependent manner (p<0.05).2. The effects of 5-aza-dC on human lung cancer cells A549Human lung cancer cells A549 were treated with 0-10 μM 5-aza-dC for 24h and 48h, respectively, results showed that compared to the untreated group, cell viability were significantly decreased in a dose-and time-dependent manner (p<0.05). Flow cytometric analysis showed that 5-aza-dC increased cell apoptosis in a dose-dependent manner (p<0.05), cells were blocked in G0/G1 stage (p<0.05), and the concerntration of reactive oxygen species in cell raised rapidly after 5-aza-dC exposure. After treated with 10 μM 5-aza-dC for 48h, the mRNA expression of relative gene was measured by real-time PCR, results showed that 5-aza-dC decreased the expression of SOX2, NANOG, cancer related gene ELPl3, Cer-bA and apoptosis related genes CASEPASE-9(p<0.05),5-aza-dC also reduce the migration and invasion ability of A549 lung cancer cell(p<0.05).In onclusion,5-aza-dC can synergistically inhibits cell proliferation in a dose-and time-dependent manner and induce cell apoptosis in bovine fetal fibroblast cells and human lung cancer cells A549,5-aza-dC caused cell cycle block and cell apoptosis through cytotoxic effectors or effect DNA methylation, the induction of cell apoptosis may be related to mitochondrial oxidatie stress or apoptosis related genes and cancer related gene expression. This study will be benefit to further explore the use of drugs on cell reprogramming, and be help to study the role of drugs in cancer treatment research.