Department of Gynecology and Obstetrics, Qingdao Women and Children's Hospital, Qingdao, China.
Eur Rev Med Pharmacol Sci. 2019 Oct;23(19):8239-8248. doi: 10.26355/eurrev_201910_19133.
The aim of this study was to clarify the potential role of PCAT1 in the occurrence and development of ovarian cancer (OC).
Expression levels of PCAT1 and NEK2 in OC tissues and cell lines were detected by quantitative Real-time polymerase chain reaction (qRT-PCR). Correlation between PCAT1 expression with tumor stage and prognosis of OC patients was analyzed. Knockdown or over-expression of PCAT1 and NEK2 were achieved by siRNA or lentivirus transfection, respectively. Subsequently, cell viability, apoptosis, cell cycle progression and migration were determined by cell counting kit-8 (CCK-8), flow cytometry and transwell assay, respectively. Furthermore, the protein levels of relative genes in Wnt pathway were detected by Western blot.
PCAT1 was highly expressed in OC tissues and cell lines, especially in tumor tissues with stage III-IV compared with stage I-II. The prognosis of OC patients with higher expression of PCAT1 was significantly worse than those with lower expression. In vitro experiments confirmed that PCAT1 knockdown obviously inhibited proliferative and migratory potentials, whereas induced apoptosis of OC cells. No significant changes were observed in cell cycle progression of OC cells after knockdown or overexpression of PCAT1. Meanwhile, overexpression of PCAT1 remarkably upregulated the expression level of NEK2, which was the target gene of PCAT1. Interestingly, NEK2 knockdown could obviously suppress cell migration. Furthermore, Western blot results elucidated that PCAT1 knockdown could inhibit the protein levels of relative genes in Wnt pathway in OC cells.
PCAT1 was highly expressed in OC tissues than adjacent normal tissues. PCAT1 overexpression significantly promoted proliferative and migratory potentials, whereas inhibited apoptosis of OC cells through upregulating NEK2 expression via Wnt pathway.
本研究旨在阐明 PCAT1 在卵巢癌（OC）发生发展中的潜在作用。
采用实时定量聚合酶链反应（qRT-PCR）检测 OC 组织和细胞系中 PCAT1 和 NEK2 的表达水平。分析 PCAT1 表达与 OC 患者肿瘤分期和预后的相关性。通过 siRNA 或慢病毒转染分别实现 PCAT1 和 NEK2 的敲低或过表达。随后，通过细胞计数试剂盒-8（CCK-8）、流式细胞术和 Transwell 测定分别测定细胞活力、凋亡、细胞周期进程和迁移。此外，通过 Western blot 检测 Wnt 通路中相关基因的蛋白水平。
PCAT1 在 OC 组织和细胞系中高表达，尤其是在 III-IV 期肿瘤组织中高于 I-II 期。PCAT1 高表达的 OC 患者的预后明显差于低表达的患者。体外实验证实，PCAT1 敲低明显抑制 OC 细胞的增殖和迁移潜能，而诱导凋亡。PCAT1 敲低或过表达后，OC 细胞的细胞周期进程无明显变化。同时，PCAT1 的过表达显著上调了 PCAT1 的靶基因 NEK2 的表达水平。有趣的是，NEK2 的敲低明显抑制了 OC 细胞的迁移。此外，Western blot 结果表明，PCAT1 敲低可抑制 OC 细胞中 Wnt 通路相关基因的蛋白水平。
PCAT1 在 OC 组织中的表达明显高于邻近正常组织。PCAT1 过表达通过上调 Wnt 通路中的 NEK2 表达，显著促进 OC 细胞的增殖和迁移潜能，而抑制凋亡。
