Department of Hepatology, the First Hospital of Jilin University, Changchun, 130021, Jilin, China.
Department of Hepatobiliary and Pancreatic Surgery, the First Hospital of Jilin University, Changchun, 130021, Jilin, China.
BMC Cancer. 2019 Jul 24;19(1):731. doi: 10.1186/s12885-019-5936-2.
Emerging evidence indicates that Long non-coding RNAs (LncRNAs) and microRNAs (miRNAs) play crucial roles in tumor progression, including hepatocellular carcinoma (HCC). However, whether there is a crosstalk between LncRNA pituitary tumor-transforming 3 (PTTG3P) and miR-383 in HCC remains unknown. This study is designed to explore the underlying mechanism by which LncRNA PTTG3P sponges miR-383 during HCC progression.
qPCR and Western blot were used to analyze LncRNA PTTG3P, miR-383 and other target genes' expression. CCK-8 assay was performed to examine cell proliferation. Annexin V-PE/PI and PI staining were used to analyze cell apoptosis and cell cycle distribution by flow cytometry, respectively. Transwell migration and invasion assays were used to examine cell migration and invasion abilities. An in vivo xenograft study was performed to detect tumor growth. Luciferase reporter assay and RNA pull-down assay were carried out to detect the interaction between miR-383 and LncRNA PTTG3P. RIP was carried out to detect whether PTTG3P and miR-383 were enriched in Ago2-immunoprecipitated complex.
In this study, we found that PTTG3P was up-regulated in HCC tissues and cells. Functional experiments demonstrated that knockdown of PTTG3P inhibited cell proliferation, migration and invasion, and promoted cell apoptosis, acting as an oncogene. Mechanistically, PTTG3P upregulated the expression of miR-383 targets Cyclin D1 (CCND1) and poly ADP-ribose polymerase 2 (PARP2) by sponging miR-383, acting as a competing endogenous RNA (ceRNA). The PTTG3P-miR-383-CCND1/PARP2 axis modulated HCC phenotypes. Moreover, PTTG3P also affected the PI3K/Akt signaling pathway.
The data indicate a novel PTTG3P-miR-383-CCND1/PARP2 axis in HCC tumorigenesis, suggesting that PTTG3P may be used as a potential therapeutic target in HCC.
新兴证据表明，长非编码 RNA（lncRNAs）和 microRNAs（miRNAs）在肿瘤进展中发挥着关键作用，包括肝细胞癌（HCC）。然而，lncRNA 垂体瘤转化基因 3（PTTG3P）与 miR-383 之间是否存在 HCC 进展过程中的相互作用尚不清楚。本研究旨在探讨 LncRNA PTTG3P 在 HCC 进展过程中通过海绵吸附 miR-383 的潜在机制。
采用 qPCR 和 Western blot 分析 lncRNA PTTG3P、miR-383 和其他靶基因的表达。CCK-8 法检测细胞增殖。通过流式细胞术分别用 Annexin V-PE/PI 和 PI 染色检测细胞凋亡和细胞周期分布。Transwell 迁移和侵袭实验检测细胞迁移和侵袭能力。进行体内异种移植研究以检测肿瘤生长。通过荧光素酶报告基因检测和 RNA 下拉实验检测 miR-383 与 LncRNA PTTG3P 之间的相互作用。RIP 实验检测 PTTG3P 和 miR-383 是否富集在 Ago2 免疫沉淀复合物中。
本研究发现 PTTG3P 在 HCC 组织和细胞中上调。功能实验表明，敲低 PTTG3P 可抑制细胞增殖、迁移和侵袭，促进细胞凋亡，发挥癌基因作用。机制上，PTTG3P 通过海绵吸附 miR-383 上调 miR-383 靶基因细胞周期蛋白 D1（CCND1）和多聚 ADP-核糖聚合酶 2（PARP2）的表达，发挥竞争性内源性 RNA（ceRNA）的作用。PTTG3P-miR-383-CCND1/PARP2 轴调节 HCC 表型。此外，PTTG3P 还影响 PI3K/Akt 信号通路。
数据表明 HCC 发生过程中存在新型 PTTG3P-miR-383-CCND1/PARP2 轴，提示 PTTG3P 可能作为 HCC 的潜在治疗靶点。
