Feng Zhaoyi, Wen Hongwu, Liao Qinping
Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing 100034, China.
Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing 100034, China. Email:
Zhonghua Fu Chan Ke Za Zhi. 2014 Oct;49(10):780-6.
To study the expression of Notch1, Jagged1 and Notch intracellular domain (NICD)in epithelial ovarian carcinoma tissues and analyze the clinical significance. To explore the activity of γ-secretase in epithelial ovarian carcinoma cell line SKOV3 and the effect of N-[N-(3, 5-dil uorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT), a γ-secretase inhibitor on the activity of γ-secretase in SKOV3.
Immunohistochemistry staining method was performed in 43 patients with epithelial ovarian carcinoma and 11 patients with benign epithelial ovarian tumor to detect the expression of Notch1, Jagged1 and NICD. The differences of expressionof Notch1, Jagged1 and NICD between malignant and benign ovarian tumors was compared and alsoanalyzed the correlation with clinicopathological parameters of ovarian carcinoma. Human serous ovarian cancer cell line SKOV3 and immortalized nontumorigenic ovarian epithelial cell line T29 were incubated in vitro. The activities of γ-secretase in SKOV3 and T29 with dimethyl sulfoxide (DMSO) and DAPT were detected respectively by Gal4-VP16/UAS and dual luciferase reporter assay system.
(1) The immunohistochemical composite scores (ICS) of Notch1 in epithelial ovarian carcinoma (6.7 ± 2.2) were not significantly different with those in benign epithelial ovarian tumor (5.4 ± 2.7, P = 0.153), while the ICS of Jagged1 and NICD in epithelial ovarian carcinoma (5.3 ± 2.4, 5.3 ± 2.3)were higher than those in benign epithelial ovarian tumor (1.6 ± 1.4, 3.1 ± 1.7; all P < 0.01). The expression of Notch1, Jagged1 and NICD had no correlation with patients' aged, history of carcinoma, ascites, the level of serum CA125, maximum length of ovarian tumor, Federation International of Gynecology and Obstetrics (FIGO) stage, grade and pathology subtypes (all P > 0.05). The hazard ratio between the high expression of Notch1, Jagged1, or NICD and the moderate to low expression of Notch1, Jagged1, or NICD, and Jagged1 were 0.771, 1.648 and 1.316, respectively (all P > 0.05). The 5-year survival rate and median survival time between the high expression of Notch, Jagged1 or NICD in subgroup and moderate to low expression in subgroup were of no difference (all P > 0.05). The activity of γ-secretase in SKOV3 was significantly higher than that in T29([ 12.2 ± 1.4)%, P = 0.019]. (2) After DAPT treated, the relative activity of γ-secretase in SKOV3 (50 µmol/L) was declined from (100.0 ± 5.3)% to(6.6 ± 0.8)% (P = 0.001).
Jagged1 and NICD in Notch1 pathway may play a key role in the occurrence of ovarian carcinoma. The activity of γ-secretase in epithelial ovarian carcinoma was higher than that in ovarian epithelial cell which suggest that DAPT, γ-secretase inhibitor, may become the target of ovarian carcinoma treatment.
研究Notch1、Jagged1及Notch胞内结构域（NICD）在上皮性卵巢癌组织中的表达情况并分析其临床意义。探讨γ-分泌酶在上皮性卵巢癌细胞系SKOV3中的活性以及γ-分泌酶抑制剂N-[N-(3,5-二氟苯乙酰基)-L-丙氨酰基]-S-苯甘氨酸叔丁酯（DAPT）对SKOV3中γ-分泌酶活性的影响。
采用免疫组织化学染色法对43例上皮性卵巢癌患者及11例良性上皮性卵巢肿瘤患者进行检测，以检测Notch1、Jagged1及NICD的表达。比较恶性和良性卵巢肿瘤中Notch1、Jagged1及NICD表达的差异，并分析其与卵巢癌临床病理参数的相关性。将人浆液性卵巢癌细胞系SKOV3和永生化非致瘤性卵巢上皮细胞系T29进行体外培养。分别采用Gal4-VP16/UAS和双荧光素酶报告基因检测系统检测SKOV3和T29中γ-分泌酶在二甲基亚砜（DMSO）和DAPT作用下的活性。
（1）上皮性卵巢癌中Notch1的免疫组织化学综合评分（ICS）为（6.7±2.2），与良性上皮性卵巢肿瘤（5.4±2.7，P = 0.153）相比无显著差异，而上皮性卵巢癌中Jagged1和NICD的ICS（5.3±2.4，5.3±2.3）高于良性上皮性卵巢肿瘤（1.6±1.4，3.1±1.7；均P < 0.01）。Notch1、Jagged1和NICD的表达与患者年龄、癌病史、腹水、血清CA125水平、卵巢肿瘤最大径、国际妇产科联盟（FIGO）分期、分级及病理亚型均无相关性（均P > 0.05）。Notch1、Jagged1或NICD高表达与中低表达之间的风险比分别为0.771、1.648和1.316（均P > 0.05）。亚组中Notch、Jagged1或NICD高表达与中低表达之间的5年生存率和中位生存时间无差异（均P > 0.05）。SKOV3中γ-分泌酶的活性显著高于T29（[12.2±1.4]%，P = 0.019）。（2）DAPT处理后，SKOV3（50 μmol/L）中γ-分泌酶的相对活性从（100.0±5.3）%降至（6.6±0.8）%（P = 0.001）。
Notch1信号通路中的Jagged1和NICD可能在卵巢癌的发生中起关键作用。上皮性卵巢癌中γ-分泌酶的活性高于卵巢上皮细胞，提示γ-分泌酶抑制剂DAPT可能成为卵巢癌治疗的靶点。
