The Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong, China.
Department of Gynecology, Jining No.1 People's Hospital, Jining 272011, Shandong, China.
Cancer Biomark. 2017 Dec 12;21(1):123-133. doi: 10.3233/CBM-170388.
Colon cancer-associated transcript 2 (CCAT2), a novel lncRNA has been reported as an oncogene in several cancers. This study was aimed to explore whether CCAT2 also exerted oncogenic roles in endometrial cancer cells.
The expression of CCAT2 in 30 pairs of endometrial cancer and matched non-cancerous tissues were detected by qRT-PCR. Two endometrial cancer cell lines HEC-1-A and RL95-2 were used throughout this study. CCAT2 in cells was silenced by transfection with shRNA targeted CCAT2, then cell growth and metastasis were assessed by performing trypan blue staining, Transwell assay, and flow cytometry. Dual-luciferase reporter assay was used to detect the combination of miR-216b and CCAT2. Besides, the expression of miR-216b and Bcl-2 in cells were overexpressed or suppressed by transfection with their correspondingly mimic/vector or inhibitor/shRNA. qRT-PCR and western blot analysis were performed to detect the expression of Bcl-2 and main factors in PTEN/PI3K/AKT and mTOR signaling pathways.
CCAT2 was highly expressed in endometrial cancer tissues when compared to non-cancerous endometrial tissues. Knockdown of CCAT2 inhibited HEC-1-A and RL95-2 cells viability, migration, invasion, but induced apoptosis. CCAT2 was an endogenous sponge by competing for miR-216b, and miR-216b suppression alleviated CCAT2 silence-diminished cell growth and metastasis. miR-216b negatively regulated Bcl-2 and Bcl-2 could further active PTEN/PI3K/AKT and mTOR signaling pathways.
To conclude, these results demonstrated lncRNA CCAT2 was highly expressed in endometrial cancer tissues. Knockdown of CCAT2 inhibited cell growth and metastasis of endometrial cancer cells by sponging miR-216b.
结肠癌相关转录物 2（CCAT2）是一种新型 lncRNA，已在多种癌症中被报道为癌基因。本研究旨在探讨 CCAT2 是否也在子宫内膜癌细胞中发挥致癌作用。
通过 qRT-PCR 检测 30 对子宫内膜癌和配对的非癌组织中的 CCAT2 表达。在整个研究过程中使用了两种子宫内膜癌细胞系 HEC-1-A 和 RL95-2。通过转染靶向 CCAT2 的 shRNA 沉默细胞中的 CCAT2，然后通过台盼蓝染色、Transwell 测定和流式细胞术评估细胞生长和转移。双荧光素酶报告基因检测用于检测 miR-216b 和 CCAT2 的结合。此外，通过转染相应的模拟物/载体或抑制剂/shRNA 来过表达或抑制 miR-216b 和 Bcl-2 的表达。通过 qRT-PCR 和 Western blot 分析检测 Bcl-2 及 PTEN/PI3K/AKT 和 mTOR 信号通路中的主要因子的表达。
与非癌性子宫内膜组织相比，CCAT2 在子宫内膜癌组织中高表达。敲低 CCAT2 抑制了 HEC-1-A 和 RL95-2 细胞的活力、迁移和侵袭，但诱导了细胞凋亡。CCAT2 通过竞争 miR-216b 作为内源性海绵，而 miR-216b 的抑制减轻了 CCAT2 沉默减弱的细胞生长和转移。miR-216b 负调控 Bcl-2，Bcl-2 可以进一步激活 PTEN/PI3K/AKT 和 mTOR 信号通路。
总之，这些结果表明 lncRNA CCAT2 在子宫内膜癌组织中高表达。敲低 CCAT2 通过海绵 miR-216b 抑制子宫内膜癌细胞的生长和转移。
