摘要
目的研究土槿乙酸衍生物PBAD1体外能否诱导宫颈癌细胞株(Hela细胞)发生凋亡及其发生机制。方法采用MTT、SRB、细胞生长曲线、细胞集落、荧光显微镜检测凋亡细胞、DNAladder、流式细胞仪等方法进行凋亡检测,用Westernblot方法检测凋亡过程中相关基因表达的变化。结果土槿乙酸衍生物PBAD1在体外试验中对Hela细胞的杀伤力强,荧光显微镜观察到了凋亡小体;DNAladder法检测到凋亡时DNA降解形成的梯带,流式细胞仪检测到了细胞凋亡峰,同时观察到细胞周期的变化。在凋亡过程中,凋亡相关基因p53基因表达明显增高,而bcl2表达下调。结论PBAD1体外能诱导Hela细胞发生凋亡,其机制可能与p53基因表达上调及bcl2基因表达下调有关。
Aim To study whether the derivation of pseudolaric acid B(PBA-D1) can induce apoptosis on human galactophore cancer Hela cells line in vitro and the possible mechanisms of apoptosis. Methods MTT,SRB assay,cell growth curve,colony formation of cancer cells, morphological study, DNA gel electrophoresis, flow cytometry,Western blot.Results Potent cytotoxic effect on human galactophore cancer Hela cells line can be observed by MTT,SRB assay and cell growth curve,colony formation of cancer cells. After treatment of Hela cells with PBA-D1, apparent morphological characteristic of apoptosis was detected under the fluorescent microscopes with the staining of Hoechst 33342, both the number of apoptosis cells and the cell circle were measured by flow cytometry, a typical “Sub-G_1 peak” was also checked and PBA-D1 blocked the tumor cells at G_2+M phase and arrested the following cell circle. Western blots showed PBA-D1 promoted p53 protein expression, at the same time it also inhibited bcl-2 expression in a dose-dependent manner.Conclusion PBA-D1 induces apoptosis by promoting p53 and inhibiting bcl-2 expression in a dose-dependent manner.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2005年第7期867-871,共5页
Chinese Pharmacological Bulletin
