lncRNA HEIH accelerates cell proliferation and inhibits cell senescence by targeting miR-3619-5p/CTTNBP2 axis in ovarian cancer
lncRNA HEIH accelerates cell proliferation and inhibits cell senescence by targeting miR-3619-5p/CTTNBP2 axis in ovarian cancer
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DOI：
10.1097/gme.0000000000001655
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发表时间：
2020-11-01
期刊：
影响因子：
--
通讯作者：
Jia, Yan
中科院分区：
文献类型：
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作者：
Si, Lihui;Chen, Jun;Jia, Yan
目的：卵巢上皮性癌是妇科最常见的恶性肿瘤之一。大量研究证实，长链非编码RNA（lncRNA）在卵巢癌中异常表达，并与肿瘤细胞的增殖和衰老密切相关。然而，卵巢癌中肝细胞癌（HEIH）长链非编码RNA高表达的作用和潜在的分子机制尚不清楚。研究方法：采用实时定量聚合酶链反应、RNA免疫沉淀、荧光素酶报告基因、荧光原位杂交、western blot、集落形成实验、β-半乳糖苷酶衰老实验、细胞凋亡、增殖、侵袭和迁移实验等方法研究HEIH在卵巢癌中的作用。数据表示为平均值+/-标准差。两组间比较采用t检验。采用单因素方差分析对多组间数据进行比较，采用Tukey事后检验。所有实验重复三次。P < 0.05为有统计学意义。结果如下：HEIH在卵巢癌组织（n = 25;比正常组织高2倍，P < 0.05）和细胞系（平均比正常卵巢上皮细胞系高6倍，P < 0.05）中的表达被发现上调，并且HEIH高表达预示不良预后（40个月后的存活率约为25%; P < 0.05）。此外，我们发现HEIH促进卵巢癌细胞的增殖、迁移和侵袭，而抑制卵巢癌细胞的衰老（P < 0.05）。在机制上，HEIH作为miR-3619- 5 p的“海绵体”，miR-3619- 5 p对HEIH介导的卵巢癌细胞增殖的调控有拮抗作用（P < 0.05）。此外，皮质素结合蛋白2（CTTNBP 2）被发现是miR-3619- 5 p的下游靶点。拯救实验证实CTTNBP 2上调显著逆转HEIH敲低对卵巢癌进展的抑制作用（P < 0.05）。此外，我们发现HEIH通过调节CTTNBP 2的表达促进体内肿瘤的生长（P < 0.05）。结论：总之，我们的研究表明，HEIH通过靶向miR-3619- 5 p/CTTNBP 2轴促进卵巢癌细胞的增殖、迁移和侵袭，而抑制细胞衰老。这些发现可能对寻找新的治疗靶点以改善卵巢癌治疗有价值。
Objective: Epithelial ovarian cancer is the most lethal malignancy in gynecology. Numerous studies have confirmed that long noncoding RNAs (lncRNAs) are abnormally expressed in ovarian cancer and are closely associated with the cell proliferation and senescence in cancers. However, the role and underlying molecular mechanism of long noncoding RNA high expression in hepatocellular carcinoma (HEIH) in ovarian cancer remain unknown. Methods: Experiments including Real-time quantitative polymerase chain reaction, RNA immunoprecipitation, luciferase reporter, Fluorescence in situ hybridization, western blot, colony formation assays, beta-galactosidase senescence assay, cell apoptosis, proliferation, invasion, and migration assays were applied to investigate the role of HEIH in ovarian cancer. The data were expressed as the mean +/- standard deviation. Student t test was used to compare the data between two groups. The one-way analysis of variance was applied to compare the data among multiple groups with Tukey post hoc test. All experiments were repeated three times. P < 0.05 was considered statistically significant. Results: Herein, HEIH expression was found to be up-regulated in ovarian cancer tissues (n = 25; twofold higher than normal tissues, P < 0.05) and cell lines (sixfold higher than normal ovarian epithelial cell line on average, P < 0.05), and high HEIH expression predicted poor prognosis (survival rate is about 25% after 40 mo; P < 0.05). Moreover, we found that HEIH accelerated proliferation, migration, and invasion, whereas inhibited cell senescence in ovarian cancer (P < 0.05). In mechanism, HEIH was confirmed to serve as a sponge for miR-3619-5p, and miR-3619-5p counteracted HEIH-mediated regulation of ovarian cancer (P < 0.05). Besides, cortactin-binding protein 2 (CTTNBP2) was found to be the downstream target of miR-3619-5p. Rescue assays validated that CTTNBP2 up-regulation significantly reversed the inhibitory effects of HEIH knockdown on ovarian cancer progression (P < 0.05). Furthermore, we found that HEIH facilitated tumor growth in vivo by regulating CTTNBP2 expression (P < 0.05). Conclusions: In conclusion, our research revealed that HEIH accelerated cell proliferation, migration and invasion, whereas inhibited cell senescence in ovarian cancer via targeting the miR-3619-5p/CTTNBP2 axis. These findings may be valuable for finding new therapeutic targets to improve ovarian cancer treatment.