Min Fengying, Chu Guoyan
Department of Gynecology, Jining No. 1 People's Hospital, Jining, Shandong, China.
Department of Gynecology and Obstetrics, Shengli Oilfield Central Hospital, Dongying, Shandong, China.
J Cell Biochem. 2020 Feb;121(2):1963-1972. doi: 10.1002/jcb.29431. Epub 2019 Oct 31.
Long noncoding RNA prostate cancer-associated transcript 1 (PCAT-1) is overexpressed in human malignancies and its silence abates the exaggeration of cancers. Whereas, the activity of PCAT-1 silence in ovarian cancer (OC) remains elusive. Here, our study was designed to corroborate the function of PCAT-1 silence in cellular activities and the molecular mechanisms. PCAT-1 in human ovarian tumor tissue specimens and cell lines (A2780 and SKOV3) were quantified by real-time quantitative reverse polymerase chain reaction (qRT-PCR). Reinforced silence of PCAT-1 and microRNA (miR)-124-3p was established by transfection and identified by qRT-PCR. The viability, apoptosis as well as migration and invasion were examined. Western blot was exploited for analysis of proteins involved in proliferation, apoptosis, migration and invasion, and signaling transduction. OC tissues showed the accumulation of PCAT-1. Silencing PCAT-1 caused the impediment of proliferation, migration, and invasion with the increase in apoptosis. PCAT-1 knockdown repressed the expression of cyclin D1, CDK6, p53, Bax, cleaved caspase-3, metallopeptidases, and vimentin with the restoration of miR-124-3p. However, the roles of PCAT-1 silence were weakened in the absence of miR-124-3p. PCAT-1 silence caused decrease in Wnt3a, β-catenin, and phosphorylation of protein kinase B and mechanistic target of rapamycin was abolished by miR-124-3p inhibitor. The tumor-suppressive role of PCAT-1 silence was mediated by miR-124-3p.
长链非编码RNA前列腺癌相关转录本1（PCAT-1）在人类恶性肿瘤中过表达，其沉默可减轻癌症的恶化。然而，PCAT-1沉默在卵巢癌（OC）中的作用仍不清楚。在此，我们的研究旨在证实PCAT-1沉默在细胞活动中的功能及其分子机制。通过实时定量逆转录聚合酶链反应（qRT-PCR）对人卵巢肿瘤组织标本和细胞系（A2780和SKOV3）中的PCAT-1进行定量。通过转染建立PCAT-1和微小RNA（miR）-124-3p的增强沉默，并通过qRT-PCR进行鉴定。检测细胞活力、凋亡以及迁移和侵袭情况。利用蛋白质印迹法分析参与增殖、凋亡、迁移和侵袭以及信号转导的蛋白质。OC组织显示PCAT-1的积累。沉默PCAT-1导致增殖、迁移和侵袭受到抑制，同时凋亡增加。PCAT-1基因敲低抑制了细胞周期蛋白D1、细胞周期蛋白依赖性激酶6、p53、Bax、裂解的半胱天冬酶-3、金属肽酶和波形蛋白的表达，同时miR-124-3p得以恢复。然而，在没有miR-124-3p的情况下，PCAT-1沉默的作用减弱。PCAT-1沉默导致Wnt3a、β-连环蛋白减少，蛋白激酶B的磷酸化和雷帕霉素靶蛋白的磷酸化被miR-124-3p抑制剂消除。PCAT-1沉默的肿瘤抑制作用是由miR-124-3p介导的。
