岗田酸促癌作用过程中对Balb/c 3T3细胞凋亡的影响

AIM  To investigate the effects of okadaic acid (OA) on the apoptosis during the promotion stage of Balb/c 3T3 cells transformation, and to discuss the involved molecular mechanisms. METHODS  The two-stage transformation test of Balb/c 3T3 cells was established with N-methyl-N′-nitro-N- nitrosoguanidine (MNNG) as initiator and OA as promoter. The cytotoxicity of OA was detected by trypan blue exclusion assay, and the cellular apoptosis was inspected by annexin V-FITC/PI staining and flow cytometry. Mouse toxicology gene chip was used to detect the gene expression of cells after OA treatment, and the expression of SPP1 gene and Txn1 gene was validated by real-time reverse transcription-polymerase chain reaction (RT-PCR) assay. RESULTS  OA 7.8 μg•L^-1 could induce the transformation of Balb/c 3T3 cells after 1 mg•L^-1 MNNG initiation. The cell viability was decreased and the percentage of apoptotic cells was increased significantly after OA treatment. In addition, OA influenced the transcriptional expression of some genes related to apoptosis or antioxidation, and the expression level of SPP1 and Txn1 genes was confirmed by real-time RT-PCR method. CONCLUSION  OA can induce apoptosis in Balb/c 3T3 cells during the promotion stage, the results suggest the possibility that the mitochondrial apoptosis pathway and oxidation stress may play important roles in the apoptosis.