Akt基因在上皮性卵巢癌组织中的表达及临床意义

目的:检测Akt基因在正常卵巢组织、上皮性卵巢囊肿、交界性上皮性卵巢肿瘤和上皮性卵巢癌组织中的表达水平,探讨其在上皮性卵巢癌演变过程中的作用及临床意义。方法:收集正常卵巢组织10例、良性上皮性卵巢囊肿组织15例、交界性上皮性卵巢肿瘤组织15例和上皮性卵巢癌组织40例。RT-PCR和Western blot法分别检测Akt mRNA和蛋白的表达水平,并分析其与临床病理特征的相关性。采用Akt激酶特异性抑制剂Triciribine分别作用于上皮性卵巢癌SKOV3、ES-2细胞,MTT法和FCM分别检测细胞增殖和凋亡,比较单独抑制剂Triciribine、单独顺铂及联合两种制剂的治疗方案对SKOV3、ES-2细胞的作用。结果:(1)Akt1、Akt2、Akt3 mRNA及p-Akt在正常卵巢组织、上皮性卵巢囊肿、交界性上皮性卵巢肿瘤和上皮性卵巢癌组织中均有表达,上皮性卵巢癌组织中的表达水平显著高于其余3组(P0.05);(2)p-Akt表达水平、总Akt mRNA水平与上皮性卵巢癌的临床分期和组织分级有关(P0.05),而与患者年龄和病理类型无关(P0.05);(3)Triciribine能有效抑制SKOV3、ES-2细胞中p-Akt水平,呈浓度依赖性抑制癌细胞生长率,联合顺铂处理可增加癌细胞的凋亡率。结论:Akt在上皮性卵巢癌组织过度激活,与临床分期和组织分级相关。Akt异常激活不仅是上游PI3K异常激活的后续效应,同时与Akt自身扩增密切相关。Triciribine阻断Akt的信号传导可增强顺铂化疗的疗效。

The expression and clinical significance of Akt in human epithelium ovarian cancer

Objective: To explore the expression and significance of Akt at protein and mRNA levels in epithelial ovarian cancer tissues.Methods: The expression of Akt at protein and mRNA levels were evaluated by Western blot and RT-PCR in normal ovarian epithelium( group N,n = 10),benign epithelial ovarian tumor tissues( group B1,n = 15),borderline epithelial ovarian tumor tissues( group B2,n = 15) and epithelial ovarian cancer tissues( group E,n = 40).The relevant clinical pathological parameters were analyzed. Western blot detected the effect of Ticiribine in SKOV3 and ES-2 cells. Methyl thiazolyl tetrazolium and flow cytometry were used to detect the growth rate and early apoptosis rate after cisplatin-based chemotherapy,Triciribine or combination treatment,respectively.Results: The tendency of two methods was coincidence on the whole.Overexpression of Akts isoforms and overactivation of Akt were detected in group E,compared with group N,group B1,and group B2( P < 0. 05). Significant differences were noted between Akt activation,clinical staging and tumor differentiation degree( P<0.05).There was no significant relationship between Akt activation and age at surgery or pathological type( P > 0.05).The activation of Akt was dramatically inhibited after exposured to 10μmol /L Triciribine under normal culture condition( P<0.05).Triciribine enhanced the sensitivity of cisplatin-based chemotherapy in SKOV3 and ES-2 cell.Conclusions: Overactivation of PI3 K /Akt signaling existed in late epithelial ovarian cancer tissues.The alteration of Akts were correlated with clinicopathological stage and tumor differentiation degree. Triciribine could enhance cisplatin-based chemotherapy in epithelial ovarian cancer cells by inhibiting phospho-Akt level.