摘要
目的探讨细胞信号转导信使磷脂酰肌醇3激酶(PI3K)在卵巢上皮性癌(卵巢癌)组织中的表达及其意义;并探讨 P13K 抑制剂 LY294002联合顺铂对卵巢癌细胞株的生长抑制作用。方法应用蛋白印迹法和 RT-PCR 技术检测正常卵巢组织(正常组,20份)、卵巢良性上皮性肿瘤组织(良性组,6份)、卵巢交界性上皮性肿瘤组织(交界性组,6份)和卵巢癌组织(卵巢癌组,39份)中P13K p85亚单位蛋白和 mRNA 的表达。将 SKOV3细胞分为对照组(只加培养液)、LY294002组(加1、10、30、50、100 μmol/L LY294002)、顺铂组(加0.33、1.25、2.5、5、10 μmol/L 顺铂)和联合组(加50 μmol/L LY294002+10 μmol/L 顺铂),四甲基偶氮唑蓝还原法测定不同浓度的 LY294002及顺铂对 SKOV3细胞的生长抑制作用。结果 (1)P13K p85亚单位蛋白的表达阳性率:正常组和良性组均为0,交界性组为2/6,卵巢癌组为85%(33/39),正常组、良性组、交界性组分别与卵巢癌组比较,差异均有统计学意义(P<0.01)。(2)P13K p85亚单位 mRNA 的表达水平:正常组为0.178±0.102,良性组为0.643±0.112,交界性组为0.847±0.058,卵巢癌组为1.689±0.423,正常组、良性组、交界性组分别与卵巢癌组比较,差异均有统计学意义(P<0.01)。卵巢癌组织中,P13K p85亚单位蛋白表达阳性率及 mRNA 的表达水平,不同年龄、病理类型间比较,差异均无统计学意义(P>0.05);而不同病理分化程度、手术病理分期间比较,差异均有统计学意义(P<0.05)。(3)LY294002及顺铂呈剂量依赖性地抑制 SKOV3细胞的生长。LY294002组(50 μmol/L)、顺铂组(10 μmol/L)、联合组 SKOV3细胞的抑制率分别为(46.0±2.0)%、(44.4±3.2)%、(57.1±4.1)%,联合组 SKOV3细胞的抑制率高于 LY294002组及顺铂组(P<0.01)。结论 P13K p85亚单位在卵巢癌组织中呈高表达,与病理分化程度、手术病理分期有关。LY294002与顺铂联合用药对卵巢癌细胞生长的抑制具有协同效应。
Objective To explore the expression and significance of phosphatidylinositol-3 kinase （PI3K） in ovarian cancer, and the effect of combined PI3K inhibitor （LY294002） therapy with cisplatin on epithelial ovarian carcinoma, and to explore if there is a synergistic effect between the two therapies. Methods The expression levels of PI3K p85 subunit proteins and mRNA were evaluated by western blot and RT-PCR in normal ovarian tissue （G1） , ovarian benign tumor tissue （G2）, ovarian borderline tumor tissue （G3） and ovarian cancer tissue （G4）, and the relevant clinical pathological parameters were analyzed. SKOV3 cells were isolated and cultured by enzymolysis method. SKOV3 cells were treated with culture medium only, LY294002 （1, 10, 30, 50, 100 μmol/L）, cisplatin （0. 33, 1.25, 2. 5, 5, 10μmol/L）, LY294002 （50 μmol/L） ＋ cisplatin （10 μmol/L） for 2 days, respectively. The effect of LY294002 and cisplatin on the growth of SKOV3 cells was measured by methyl thiazolyl tetrazolium assay. Retmits There was no positive expression of PI3K p85 subunit proteins in G1 and G2, while the expression was 2/6 in G3, and 85% （33/39） in G4. PI3K p85 subunit mRNA expression levels were 0. 178 ＋0. 102 inG 1, 0.643 ±0.112 in G2, 0.847 ±0.058 in G3, 1.689 ±0.423 in G4; there was a significant difference between G1, G2, G3 and G4 （ P 〈 0. 01 ）. There was no significant correlation between protein expre^ion and age at surgery or clinico-pathological staging （ P 〉 0.05 ）. Significant differences were noted between protein expression levels in G4 （ Ⅲ, Ⅳ ） and G4 （ Ⅰ , Ⅱ ; P 〈 0. 05 ）. There was a significant difference between expression levels in tissues of different differentiation degrees （ P 〈 0.05 ）. LY294002 and eisplatin inhibited the growth of SKOV3 cell in a concentration-dependent manner. The inhibitory activity of LY294002 at the concentration of 50 μmol/L was （46. 0 ± 2.0）% after treatment for two days. The inhibitory activity of cisplati
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2007年第3期196-200,共5页
Chinese Journal of Obstetrics and Gynecology
基金
上海市卫生局科技发展基金(054064)
