土槿乙酸衍生物PB-LY体外抗肿瘤作用及其机制研究

目的通过研究土槿乙酸衍生物PB-LY对肿瘤细胞的影响,探讨PB-LY的抗肿瘤活性及其作用机制。方法采用MTT法和绘制细胞生长曲线等方法检测PB-LY对多种肿瘤细胞的抑制作用;采用荧光染色、DNAladder和流式细胞术等方法检测PB-LY诱导肿瘤细胞凋亡的作用;采用RT-PCR法检测PB-LY对肿瘤细胞相关凋亡基因Caspase-3、Bcl-2和Bax的mRNA表达的影响。结果PB-LY作用于多种肿瘤细胞的IC50在1.35μmol.L-1～3.68μmol.L-1之间,且有明显的量效关系,能够诱发肿瘤细胞产生凋亡小体、DNAladder和细胞凋亡峰,上调相关促凋亡基因Caspase-3和Bax的表达,同时协同下调抑凋亡基因Bcl-2的表达。结论PB-LY能抑制肿瘤细胞的生长,同时影响肿瘤细胞内相关促凋亡和抑凋亡基因的表达,从而诱导肿瘤细胞发生凋亡。

Antitumor activity of PB-LY and its mechanism in vitro

Aim To determine the antitumor activity of PB-LY and investigate its anticancer mechanism in vitro.Methods MTT and cell growth curve were used to study antitumor activity of PB-LY.Morphological study,DNA gel electrophoresis and flow cytometric analysis were used to determine tumor cell apoptosis induced by PB-LY.RT-PCR investigated the expression of Caspase-3 mRNA,Bax mRNA and Bcl-2 mRNA in Hela cells exposed to PB-LY.Results MTT showed that IC50 was 1.35 μmol·L-1～3.68 μmol·L-1.After tumor cells were treated with PB-LY,morphological characteristic of apoptosis was detected.Agarose gel electrophoresis showed typical DNA ladder.Flow cytometric analysis showed apoptotic "Sub-Gl peak".RT-PCR showed that Caspase-3 mRNA and Bax mRNA levels increased,meanwhile the expression of Bcl-2 mRNA decreased.Conclusion PB-LY exerted obvious anticancer activity,which induced apoptosis by upregulating Caspase-3 and Bax,and downregu1ating Bcl-2.