摘要
目的:探讨三磷酸肌醇(IP3)和survivin蛋白表达变化在genistein诱导肝癌细胞凋亡中的作用。方法:以肝癌HepG2细胞培养72h为对照组,实验各组以60μmol/L的genistein作用于HepG2细胞不同时间后,应用同位素试剂盒检测细胞IP3含量,Western blotting分析细胞survivin蛋白表达,流式细胞仪检测细胞凋亡率。结果:对照组IP3含量为(29·2±0·6)pmol/106cells,survivin蛋白与内参β-actin蛋白灰度和面积之积的比值V-survivin/V-β-actin为0·63±0·06;细胞凋亡率为(2·6±0·1)%。Genistein作用于肝癌HepG2细胞12h、24h、48h、72h后IP3分别为(12·0±1·4)pmol/106cells、(7·5±0·8)pmol/106cells、(5·6±0·5)pmol/106cells、(3·3±0·6)pmol/106cells,与对照组(29·2±0·6)pmol/106cells比,P<0·01。V-survivin/V-β-actin分别为0·36±0·13、0·33±0·03、0·23±0·04、0·18±0·04,与对照组(0·63±0·06)比,P<0·01。细胞凋亡率分别为(2·7±0·2)%、(7·4±0·5)%、(20·5±2·0)%、(30·7±1·6)%,与对照组(2·6±0·1)%比,P<0·01。结论:Genistein能减少IP3生成,下调survivin基因表达,诱导肝癌细胞凋亡。
AIM- To probe into the role of 1, 4, 5 - trisphosphate inositol （IP3） and survivin protein in apoptosis of HepG2 cells induced by genistein. METHODS： HepG2 cells were treated with 60 μmol/L genistein for 12 h, 24 h, 48 h and 72 h. IP3, survivin and apoptosis rate were assayed by IP3 - [^3H] Birtrak assay, Western blotting and flow cytometry, respectively. RESULTS： IP3 in groups incubated for 12 h, 24 h, 48 h and 72 h with 60μmol/L genistein were significantly lower than that in control （P〈0.01） [（12.0±1.4） pmol/10^6cells, （7.5±0.8） pmol/10^6 cells, （5.6±0.5） pmol/10^6cells, （3.3±0.6） pmol/ 10^6 cells, vs （29.2±0.6） pmol/10^6 cells]. V- survivin/V-β- actin, which was the gray degree multiply area of survivin/the gray degree multiply area of β - actin in groups incubated for 12 h, 24 h, 48 h and 72 h with 60 μmol/L genistein, were significantly lower than that in control （ P 〈 0.01 ） [ （0.36 ± 0.13, 0.33 ± 0.03, 0.23± 0.04, 0.18 ± 0.04）, vs 0.63 ± 0.06]. The apoptosis rate in groups incubated with 60 μmol/L genistein for 24 h, 48 h and 72 h was significantly higher than that in control （ P 〈 0.01） [（7.4% ±0.5%, 20.5% ± 2.0%, 30.7% ± 1.6%） vs 2.6% ±0.1%]. CONCLUSION： Genistein induces apoptesis in HepG2 cells by reducing IP3 production and survivin protein expression.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第2期372-375,共4页
Chinese Journal of Pathophysiology
