摘要
为研究稀土化合物抗癌作用及其机制,本文采用差异蛋白质组学方法研究钆对宫颈癌细胞(HeLa)的作用,以四噻唑蓝比色法(MTT法)检测柠檬酸钆配合物([Gd(Cit)2]3-)对HeLa细胞生长的影响,并用Hoechst33258染色法观察细胞凋亡形态,利用双向凝胶电泳技术分离[Gd(Cit)2]3-作用HeLa细胞前后的蛋白,经图像分析后对差异点,进行MALDI-TOF-MS鉴定和数据检索。结果表明0.005～1mmol.L-1浓度范围内的[Gd(Cit)2]3-对HeLa细胞生长的影响存在浓度依赖性。Hoechst33258染色结果表明0.07mmol.L-1[Gd(Cit)2]3-作用24h对HeLa细胞的生长的抑制率为34%。质谱共鉴定出12个差异表达蛋白质点,包括延伸因子2、过氧化酶6、亲环蛋白A、烯醇酶1等在细胞中分别参与蛋白质转录与翻译、氧化应激、信号传导等过程的蛋白质。提示[Gd(Cit)2]3-可能通过调节胞内氧化应激水平,介导与调控信号转导与蛋白质表达等,从而诱导HeLa癌细胞凋亡。
To study the mechanism of anti-cancer actions of lanthanide compounds, proteomic experiments were performed to study the effects and mechanism of gadolinium on cervical cancer cell line HeLa. MTT assay was used to detect the viability of cells exposed to gadolinium in the form of [ Gd（ Cit）2 ] ^3- at different concentrations. The cell apoptosis induced by [ Gd （Cit） 2 ]^ 3- was observed by Hoeehst 33258 staining method. Total proteins were extracted from the exposed cells and applied to the two-dimensional gel electrophoresis for protein separation. The MALDITOF-MS was used to identify the differentially expressed proteins from gadolinium citrate-treated cells and the control. The results showed that 0. 005 -1 mmol·L^-1 of [ Gd （Cit） 2 ]^ 3- had negative correlation with the proliferation of HeLa cells after treated for 24 h. The observation from Hoechst 33258 staining indicated that 0.07 mmol· L^-1 [ Gd （Cit） 2 ] ^3- could significantly inhibit cell proliferation at an inhibitory rate of 34%. The results from two dimensional gel electrophoresis and MALDI-TOF mass spectrometry revealed 12 proteins expressed differentially in the gels of GdⅢ-treated proteins over the control, including elongation factor 2, peroxiredoxin 6, cyclophilin A, enolase 1 et al. These proteins played important roles in the processes of transcription and translation, oxidative stress responses, and signal transduction. Based on these results, the mechanism of [ Gd （Cit）2]^3- inducing the apoptosis of HeLa cells was discussed.
出处
《中国稀土学报》
CAS
CSCD
北大核心
2009年第3期435-440,共6页
Journal of the Chinese Society of Rare Earths
基金
国家自然科学基金重点项目(20637010)资助
