摘要
目的探讨genistein对肝癌HepG2细胞株PTEN和survivin蛋白表达的影响及在诱导凋亡中的作用。方法以肝癌HepG2细胞培养72 h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PTEN和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／106 cells,PTEN蛋白与内参Actin蛋白灰度和面积之积的比值V-PTEN／V-actin为0．12±0．001, survivin蛋白与内参actin蛋白灰度和面积之积的比值V-survivin／V-actin为0．63±0．06,细胞凋亡率为2．6％±0．1％。Genistein作用于肝癌Hep G2细胞12 h、24 h、48 h、72 h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; V-PTEN／V-actin分别为13．13±0．20、19．17±1．09、28．51±2．18、41．12±3．80;V-survivin／V-actin分别为0．36±0．13、0．33±0．03、0．23±0．04、0．18±0．04;细胞凋亡率分别为2．7％±0．2％、7．4％±0．5％、20．5％±2．0％、30．7％±1．6％。与对照组相比,genistein作用于肝癌Hep G2细胞12 h后各时相IP3和survivin蛋白显著降低,24 h后各时相细胞凋亡率显著增高。结论Genistein能减少IP3生成,上调PTEN基因表达,下调survivin基因表达,诱导肝癌细胞凋亡。
Objective To study the effect of inhibiting phosphate inositol signal pathway by genistein on PTEN and survivin gene expression in hepatocellular carcinoma HepG2 cells. Methods HepG2 cells were treated with 60 μmol/L genistein for 12 h, 24 h,48 h and 72 h. 1,4,5-trisphosphate inositol （ IP3 ） was measured by IP3-H^3 Birtrak Assay. PTEN and survivin protein was assayed by Westernblotting. Apoptosis rate was assayed by flow cytometry. Results IP3 were 29. 2 ± 0. 6 pmol/10^6 cells in control, 12.0 ±1.4 pmol/10^6 cells, 7.5 ± 0. 8 pmol/10^6 cells, 5.6 ± 0. 5 pmol/10^6 cells, and 3.3 ± 0. 6 pmol/10^6 cells in HepG2 cells incubated for 12 h, 24 h, 48 h and 72 h with 60 μmol/L genistein, all P 〈0. 01 compared with control. V-PTEN/V-actin （ gray degree multiply area of PTEN over the gray degree multiply area ofactin）, was 0.12 ±0.001 in control, 13. 13±0.20, 19. 17 ± 1.09, 28.51 ±2.18 and 41.12 ± 3.80 in HepG2 cells incubated with 60 μmol/L genlstein for 12 h, 24 h, 48 h and 72 h, respectively （all P 〈 0. 01 compared with control）. V-survivin/ V-actin was 0. 62 ± 0. 06 in control, 0. 36 ± 0. 13, 0. 33 ±0. 03, 0. 23±0. 04 and 0. 18 ±0. 04 in cells incubated for 12 h, 24 h, 48 h and 72 h with 60 μmol/ Lgenistein （all P〈0.01）. The apoptosis rate was 2. 6%±0.1% in control, 2.7% ±0.2%, 7.4% ± 0.5%, 20. 5% ± 2. 0% and 30. 7% ±1.6% in corresponding cell groups （all P 〈 0.01 ）. Conclusion Genistein induces apoptosis of HepG2 cells by upregulating PTEN protein expression and downregulating survivin protein expression,
出处
《中华普通外科杂志》
CSCD
北大核心
2006年第9期654-656,共3页
Chinese Journal of General Surgery
