Hu Hongzhuan, Liu Zhonglan, Liu Cun
Department of Obstetrics, Shouguang People's Hospital, Weifang, Shandong 262700, P.R. China.
Department of Gynaecology, The People's Hospital of Qihe County, Qihe, Shandong 251100, P.R. China.
Oncol Lett. 2019 Mar;17(3):2788-2794. doi: 10.3892/ol.2019.9896. Epub 2019 Jan 7.
Correlation of osteopontin (OPN) gene expression with proliferation and apoptosis of ovarian cancer cells and prognosis of patients was investigated. The expression levels of OPN in 81 pairs of ovarian cancer tissues and para-carcinoma tissues obtained via surgical resection were detected using immunohistochemistry (IHC). The correlation of OPN protein expression with clinicopathological features of patients was analyzed. All patients were followed up for 3 years. The disease-free survival (DFS) and overall survival (OS) curves of patients in high/low OPN expression groups were drawn using the Kaplan-Meier method. The expression levels of OPN in normal ovarian epithelial IOSE80 cells and 5 ovarian cancer cell lines were detected via western blotting. Moreover, two cell lines with high OPN expression were interfered with lentiviral transfection technique. The effects of OPN on ovarian cancer cell proliferation and apoptosis were detected and analyzed via Cell Counting Kit-8 (CCK8) assay and flow cytometry. The positive expression rate of OPN protein in tumor tissues was higher than that in para-carcinoma tissues (P<0.05). Survival curves suggested that both DFS and OS in OPN negative group were superior to those in OPN positive group (P<0.05). Results of western blotting showed that OPN was weakly expressed in IOSE80 cells, whereas it was highly expressed in SKOV-3, COC1, A2780, HO-8910 and OVCAR-3 cells, among which the OPN protein expression levels were relatively higher in SKOV-3 and OVCAR-3 cell lines. After knockdown of OPN gene with sh-OPN, the cell proliferation rates of OVCAR-3 and SKOV-3 were significantly decreased from 48 h (P<0.05), but the apoptosis level was increased remarkably (28.2 vs. 1.3% and 25.3 vs. 3.2%), and differences were statistically significant (P<0.05). In conclusion, overexpression of OPN enhances the proliferation of ovarian cancer cells, which is an adverse factor for patient survival and prognosis.
研究骨桥蛋白（OPN）基因表达与卵巢癌细胞增殖、凋亡及患者预后的相关性。采用免疫组织化学（IHC）检测81对手术切除获得的卵巢癌组织和癌旁组织中OPN的表达水平。分析OPN蛋白表达与患者临床病理特征的相关性。所有患者随访3年。采用Kaplan-Meier法绘制高/低OPN表达组患者的无病生存期（DFS）和总生存期（OS）曲线。通过蛋白质免疫印迹法检测正常卵巢上皮IOSE80细胞和5种卵巢癌细胞系中OPN的表达水平。此外，采用慢病毒转染技术干扰两种OPN高表达细胞系。通过细胞计数试剂盒-8（CCK8）法和流式细胞术检测并分析OPN对卵巢癌细胞增殖和凋亡的影响。肿瘤组织中OPN蛋白阳性表达率高于癌旁组织（P<0.05）。生存曲线表明，OPN阴性组的DFS和OS均优于OPN阳性组（P<0.05）。蛋白质免疫印迹结果显示，OPN在IOSE80细胞中弱表达，而在SKOV-3、COC1、A2780、HO-8910和OVCAR-3细胞中高表达，其中SKOV-3和OVCAR-3细胞系中OPN蛋白表达水平相对较高。用sh-OPN敲低OPN基因后，OVCAR-3和SKOV-3细胞的增殖率从48小时起显著降低（P<0.05），但凋亡水平显著升高（分别为28.2%对1.3%和25.3%对3.2%），差异有统计学意义（P<0.05）。总之，OPN的过表达增强了卵巢癌细胞的增殖，这是患者生存和预后的不利因素。
