Downregulation of miR-575 Inhibits the Tumorigenesis of Gallbladder Cancer via Targeting p27 Kip1
Downregulation of miR-575 Inhibits the Tumorigenesis of Gallbladder Cancer via Targeting p27 Kip1
复制标题
下调 miR-575 通过靶向 p27 Kip1 抑制胆囊癌的肿瘤发生
DOI：
10.2147/ott.s229614
复制
发表时间：
2020-01-01
影响因子：
4
通讯作者：
Fu, Yang
中科院分区：
文献类型：
--
作者：
Qin, Yiyu;Mi, Wunan;Fu, Yang
背景：胆囊癌是世界上最常见的胆道恶性肿瘤。据报道，microRNA-575 （miR-575）参与了许多癌症的肿瘤发生。然而，miR-575在GBC进展中的作用在很大程度上仍然未知。方法：采用实时定量聚合酶链反应检测miR-575在GBC细胞中的表达。CCK-8法和Ki-67染色法检测GBC细胞的增殖情况。流式细胞术检测GBC细胞凋亡，transwell法检测细胞侵袭。此外，采用Western blot方法检测GBC细胞的蛋白表达。使用Targetscan和miRDB预测miR-575的靶基因。最后，建立异种移植肿瘤模型，验证miR-575在GBC体内的功能。结果：我们的研究结果表明，miR-575拮抗剂可以降低GBC细胞的增殖和侵袭。此外，miR-575拮抗剂通过诱导G1阻滞显著诱导GBC细胞凋亡。同时，通过荧光素酶报告基因实验发现p27 Kip1是miR-575的直接靶点。此外，miR-575拮抗剂显著降低了GBC细胞中CDK1和cyclin E1的表达，上调了cleaved caspase3和p27 Kip1的水平。最后，miR-575拮抗剂在体内显著抑制GBC肿瘤生长。结论：下调miR-575通过靶向p27 Kip1显著抑制GBC的肿瘤发生。因此，miR-575可能是治疗GBC的潜在新靶点。
Background: Gallbladder cancer (GBC) is the most common biliary tract malignant cancer worldwide. It has been reported that microRNA-575 (miR-575) was involved in the tumorigenesis of many cancers. However, the role of miR-575 during the progression of GBC remains largely unknown.Methods: The expression of miR-575 in GBC cells was detected by quantitative real-time polymerase chain reaction. The proliferation of GBC cells was examined by CCK-8 assay and Ki-67 staining. Apoptosis of GBC cells was measured by flow cytometry, and cell invasion was tested by transwell assay. Moreover, protein expressions in GBC cells were evaluated using Western blot. The target gene of miR-575 was predicted using Targetscan and miRDB. Finally, xenograft tumor model was established to verify the function of miR-575 in GBC in vivo.Results: Our findings indicated that miR-575 antagonist decreased the proliferation and invasion of GBC cells. In addition, miR-575 antagonist significantly induced apoptosis of GBC cells via inducing G1 arrest. Meanwhile, p27 Kip1 was found to be a direct target of miR-575 with luciferase reporter assay. Moreover, miR-575 antagonist significantly decreased the expressions of CDK1 and cyclin E1 and upregulated the levels of cleaved caspase3 and p27 Kip1 in GBC cells. Finally, miR-575 antagonist notably suppressed GBC tumor growth in vivo.Conclusion: Downregulation of miR-575 significantly inhibited the tumorigenesis of GBC via targeting p27 Kip1. Thus, miR-575 might be a potential novel target for the treatment of GBC.