桥接整合因子1在上皮性卵巢癌组织和细胞中的表达及其临床意义

Expression and clinical significance of bridging intergrator 1 in epithelial ovarian cancer tissues and cells

目的:探讨桥接整合因子1(BIN1)在上皮性卵巢癌(EOC)组织中的表达及其临床意义,以及BIN1对EOC细胞A2780增殖、迁移和侵袭的影响。方法:收集2017年7月至2018年1月河北医科大学第四医院手术切除的67例EOC患者的肿瘤组织及同期因其他妇科疾病手术切除的30例非肿瘤患者的卵巢组织(正常对照组)标本。用免疫组织化学染色法检测EOC组织和非肿瘤卵巢组织中BIN1蛋白的表达水平,χ;检验分析BIN1表达与患者临床病理特征之间的关联,Kaplan-Meier法分析BIN1表达与患者的无病生存期(DFS)和总生存期(OS)之间的关系。用qPCR和WB法检测EOC细胞SKOV3、A2780和人卵巢上皮细胞IOSE80中BIN1 m RNA和蛋白的表达水平。利用基因转染技术将BIN1质粒CMV-MCS-GFP-SV40-NeomycinBIN1和空载体质粒CMV-MCS-GFP-SV40-Neomycin分别转染到A2780细胞以构建过表达BIN1细胞及其对照,用qPCR和WB法分别检测转染细胞中BIN1 mRNA和蛋白的表达水平,CCK-8、划痕愈合和Transwell实验分别检测过表达BIN1对A2780细胞增殖、迁移和侵袭的影响。结果:EOC组织中BIN1阳性表达率显著低于正常卵巢组织(P<0.01)。BIN1表达与EOC患者较晚的术后病理分期、较差的组织学分级、淋巴结转移及腹膜转移存在正向关联(均P<0.05);BIN1低表达组患者的DFS和OS均短于BIN1高表达组患者(均P<0.05)。SKOV3和A2780细胞中BIN1 mRNA和蛋白的表达水平均显著低于IOSE80细胞(均P<0.01);过表达BIN1显著抑制A2780细胞的增殖、迁移和侵袭(P<0.05或P<0.01)。结论:BIN1在EOC组织和细胞中呈低表达状态,与患者的不良预后有关;过表达BIN1可降低EOC细胞A2780的增殖、迁移和侵袭能力。

Objective: To investigate the expression and clinical significance of bridging intergrator 1(BIN1) in epithelial ovarian cancer(EOC) tissues, and to explore its effect on the proliferation, migration and invasion of ovarian cancer A2780 cells. Methods: The tumor tissues of 67 patients with EOC who underwent tumor resection in the Fourth Hospital of Hebei Medical University from July2017 to January 2018 were collected. Ovarian tissues resected from 30 non-tumor patients with other gynecological diseases during the same period were collected as normal controls. Immunohistochemical staining was used to detect the expression of BIN1 in EOC tissues and non-tumor ovarian tissues. χ;test was used to analyze the correlation between the expression of BIN1 and various clinicopathological factors of EOC patients. Kaplan-Meier method was used to analyze the correlation between the expression of BIN1 and disease-free survival(DFS) or overall survival(OS) of patients. The mRNA and protein expression levels of BIN1 in EOC cells(SKOV3 and A2780) and human ovarian epithelial IOSE80 cells were detected by qPCR and WB. The BIN1 plasmid CMV-MCS-GFPSV40-neomycin-BIN1 and the empty plasmid CMV-MCS-GFP-SV40-Neomycin were respectively transfected into A2780 cells to construct BIN1 overexpressed cells and its control. The mRNA and protein expression levels of BIN1 in transfected cells were detected by q PCR and WB, respectively. The effects of BIN1 overexpression on the proliferation, migration and invasion of A2780 cells were detected by CCK-8 test, wound-healing assay and Transwell test, respectively. Results: The positive expression rate of BIN1 in EOC tissues was significantly lower than that in normal ovarian tissues(P<0.01). Low BIN1 expression was positively related with advanced postoperative pathological stage, worse histological grade, lymph node metastasis and peritoneal metastasis in patients with EOC(all P<0.05). The DFS and OS of the patients in the low BIN1 expression group were shorter than those in the high BIN1 expression group(all P<0.05). The m RNA and protein expression levels of BIN1 in SKOV3 and A2780 cells were significantly lower than those in IOSE80 cells(all P<0.01). Overexpression of BIN1 significantly inhibited the proliferation, migration and invasion of A2780 cells(P<0.05 or P<0.01). Conclusion: BIN1 is lowly expressed in EOC tissues and cells, and its low expression is related to the poor prognosis of EOC patients. Overexpression of BIN1 can inhibit the proliferation, migration and invasion ability of ovarian cancer A2780 cells.