Li Jie, Huang He, Li Yingguang, Li Li, Hou Wenhui, You Zeshan
Department of Gynecology, The Eastern Hospital of the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510700, P.R. China.
Department of Gynecology, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong 510000, P.R. China.
Oncol Rep. 2016 Dec;36(6):3241-3250. doi: 10.3892/or.2016.5200. Epub 2016 Oct 24.
Long non-coding RNA growth arrest-specific 5 (GAS5) was reported to be aberrantly expressed in various types of cancers. However, the role of GAS5 in the evolution and progression of ovarian cancer remains elusive. In the present study, we aimed to investigate the cellular function and clinical significance of GAS5 in ovarian cancer. GAS5 expression levels in 63 ovarian cancer tissues were detected by quantitative real-time PCR. Cell Counting Kit-8 (CCK-8) assay was performed to analyze the effect of GAS5 on cell proliferation. The effect of GAS5 on cell migration and invasion was detected using Transwell assay. Cell apoptosis was evaluated by flow cytometry and Hoechst staining. SKOV3 cells with stable expression of GAS5 were injected into nude mice to study the effect of GAS5 on tumorigenesis in vivo. Western blotting was used to determine the protein levels of GAS5 potential targets. The results showed that GAS5 was markedly decreased in tumor tissues and a lower expression of GAS5 was detected in tumors with larger size, deeper invasive depth and higher tumor stage. Patients with low GAS5 expression level had poorer disease-free (P<0.0001) and overall survival (P=0.0016) than those with high GAS5 expression. Moreover, overexpression of GAS5 was demonstrated to suppress ovarian cancer cell proliferation in vitro and in vivo. Finally, we found that GAS5 influenced ovarian cancer cell proliferation, partly via regulating cyclin D1, p21 and apoptosis protease activating factor 1 (APAF1) expression. Our findings suggest that lncRNA GAS5 may represent a novel indicator of poor prognosis in ovarian cancer and may be a potential therapeutic target for diagnosis and therapy.
据报道,长链非编码RNA生长停滞特异性5(GAS5)在多种类型的癌症中表达异常。然而,GAS5在卵巢癌发生发展中的作用仍不清楚。在本研究中,我们旨在探讨GAS5在卵巢癌中的细胞功能及临床意义。通过定量实时PCR检测63例卵巢癌组织中GAS5的表达水平。采用细胞计数试剂盒-8(CCK-8)法分析GAS5对细胞增殖的影响。使用Transwell法检测GAS5对细胞迁移和侵袭的影响。通过流式细胞术和Hoechst染色评估细胞凋亡。将稳定表达GAS5的SKOV3细胞注射到裸鼠体内,研究GAS5对体内肿瘤发生的影响。采用蛋白质印迹法测定GAS5潜在靶点的蛋白水平。结果显示,肿瘤组织中GAS5明显降低,在肿瘤体积较大、浸润深度较深和肿瘤分期较高的肿瘤中检测到较低的GAS5表达。GAS5表达水平低的患者无病生存期(P<0.0001)和总生存期(P=0.0016)均比GAS5表达高的患者差。此外,GAS5过表达在体外和体内均能抑制卵巢癌细胞增殖。最后,我们发现GAS5部分通过调节细胞周期蛋白D1、p21和凋亡蛋白酶激活因子1(APAF1)的表达影响卵巢癌细胞增殖。我们的研究结果表明,lncRNA GAS5可能是卵巢癌预后不良的一个新指标,可能是诊断和治疗的潜在靶点。