Wang Xuegang, Wang Rong, Wu Zhun, Bai Peide
1Department of Urology, The First Affiliated Hospital of Xiamen University, The First Clinical College of Fujian Medical University, Xiamen, 361003 Fujian People's Republic of China.
2Department of Urology, The Jintan Hospital Affiliated with Jiangsu University, Changzhou, 213200 Jiangsu People's Republic of China.
Cancer Cell Int. 2019 Dec 3;19:328. doi: 10.1186/s12935-019-0994-8. eCollection 2019.
Circular RNA Itchy E3 ubiquitin protein ligase (Circ-ITCH) is significantly down-regulated in various kinds of tumors, however, the mechanisms of action and functions of circITCH gene in prostate cancer (PC) are still under investigation. The mail goal of this research was to study the functional role of Circ-ITCH gene in prostate cancer and to illuminate the function role of circ-ITCH gene in prostate cancer by targeting miR-17-5p/HOXB13.
RT-qPCR was applied to measure the expression level of circ-ITCH and miR-17-5p in PC cell lines and tissues. CCK-8, colony formation, Brdu incorporation labeling and flow cytometry assays were applied to detect the effects of circ-ITCH and miR-17-5p on proliferation and cell apoptosis. Target gene prediction and screening, luciferase reporter gene assays were utilized to assess downstream target genes of miR-17-5p and Circ-ITCH. The protein and expression of HOXB13 gene were measured by Western blotting and RT-qPCR.
CircITCH was significantly reduced in PC cell lines and tissues. Low circITCH expression level was highly related with preoperative PSA, tumor stage and Gleason score. Overexpression of circITCH can inhibit the malignant phenotype of prostate cancer. There was a high negative relationship between the expression level of microRNA-17-5p and circITCH in PC tissues, however, there existed a positive relationship between the expression of HOXB13 and circITCH. CircITCH acted as a sponge of miR-17-5p to increase HOXB13 gene expression. In addition, miR-17-5p overexpression or HOXB13 silencing can reduce the carcinogenic effects of circICCH in prostate cancer.
CircITCH promoted prostate cancer progression by regulating the HOXB13/miR-17-5p axis, and circITCH have a potential usage as therapeutic target for PC tumors.
环状RNA瘙痒E3泛素蛋白连接酶(Circ-ITCH)在各类肿瘤中显著下调,然而,circITCH基因在前列腺癌(PC)中的作用机制和功能仍在研究中。本研究的主要目的是探讨Circ-ITCH基因在前列腺癌中的功能作用,并通过靶向miR-17-5p/HOXB13阐明circ-ITCH基因在前列腺癌中的功能作用。
应用RT-qPCR检测circ-ITCH和miR-17-5p在PC细胞系和组织中的表达水平。采用CCK-8、集落形成、Brdu掺入标记和流式细胞术检测circ-ITCH和miR-17-5p对增殖和细胞凋亡的影响。利用靶基因预测和筛选、荧光素酶报告基因检测评估miR-17-5p和Circ-ITCH的下游靶基因。通过蛋白质印迹法和RT-qPCR检测HOXB13基因的蛋白和表达情况。
CircITCH在PC细胞系和组织中显著降低。低circITCH表达水平与术前PSA、肿瘤分期和Gleason评分高度相关。circITCH的过表达可抑制前列腺癌的恶性表型。PC组织中微小RNA-17-5p与circITCH的表达水平呈高度负相关,而HOXB13的表达与circITCH呈正相关。CircITCH作为miR-17-5p的海绵,增加HOXB13基因表达。此外,miR-17-5p过表达或HOXB13沉默可降低circICCH在前列腺癌中的致癌作用。
CircITCH通过调节HOXB13/miR-17-5p轴促进前列腺癌进展,circITCH作为PC肿瘤的治疗靶点具有潜在应用价值。