化学合成2’-氧-甲基鸟嘌呤核苷(P6)诱导HeLa细胞凋亡及其机制的研究

The apoptosis-inducing effects of a novel synthetic nucleoside 2'-O-meth-ylguanosine(P6) on HeLa cell

研究化学合成2’-氧-甲基鸟嘌呤核苷(P6)在体外诱导HeLa细胞凋亡的作用,观察其是否同天然提取的P6具有相同的抗肿瘤作用。首先采用形态学观察、MTT实验及流式细胞术的方法观察P6对HeLa细胞的增殖抑制作用。然后采用DNA Ladder检测、流式细胞术、RT-PCR技术及化学发光检测技术观察P6对HeLa细胞的促凋亡作用。结果表明,化学合成的P6可以抑制HeLa细胞的增殖,机制是由于细胞被阻滞于S期。化学合成的P6也可以促进HeLa细胞的凋亡,其机制在于下调了抗凋亡基因Bcl-2的转录,并促进了Caspase-3的激活。因而化学合成的P6有望成为一种新的抗肿瘤药。

To investigate the apoptosis-inducing effect of a novel synthetic nucleotide-2＇-O-methylgranosine（P6） on human cervical carcinoma cell line HeLa cell in vitro, in order to determine whther it has the same antitumor activity as the natural apoptosis-inducing nucleotide（AIN）, the proliferation inhibition effect of P6 was evaluated by means of microscopical examination, MTT assay and flow cytometery analysis, and its apoptosis-inducing effect was determined by DNA fragmentation detection,flow cytometry, RT-PCR analysis and chemiluminescence assay. The results showed that the synthetic compound P6 could inhibit the proliferation of HeLa cells by arresting these cells in the S-phase of cell cycle and by down-regulating the transcription of the anti-apoptotic Bcl-2 gene, it could induce apoptosis of HeLa cells, and promote the activation of Caspase-3. All these results suggest that the synthetic nucleotide P6 has the potential attempted to be used as an anti-tumor agent in the future.