miR-520d-5p 通过靶向 PTK2 在宫颈癌中发挥肿瘤抑制基因的作用。

MiR-520d-5p functions as a tumor-suppressor gene in cervical cancer through targeting PTK2.

机构信息

Department of Gynecology, The Maternal and Child Health Hospital of Zibo City, Zibo City, Shandong 255029, China.

Department of Emergency, The Maternal and Child Health Hospital of Zibo City, Zibo City, Shandong 255029, China.

出版信息

Life Sci. 2020 Aug 1;254:117558. doi: 10.1016/j.lfs.2020.117558. Epub 2020 Mar 17.

DOI:10.1016/j.lfs.2020.117558

PMID:32198053

Abstract

OBJECTIVE

PTK2 has been reported to be involved in tumor progression, but its regulating mechanisms in cervical cancer (CC) remain to be elusive. MiRNA-520d-5p was demonstrated to regulate the expression of many genes and inhibit the development of human tumors. However, the functional mechanisms of miRNA-520d-5p in the regulation of cervical cancer are not fully understood.

METHODS

RT-qPCR was employed to detect the expression levels of miR-520d-5p and PTK2. Western blot was performed to detect the expression levels of proteins. Dual-luciferase reporter assay was utilized to investigate the associations between miR-520d-5p and PTK2. CCK-8 assay was carried out to measure cell proliferation. In addition, transwell assay and scratch assay were used for cell invasion and migration analysis. Flow cytometry was used to detect cell apoptosis of cervical cancer.

RESULTS

The expression levels of PTK2 were elevated in CC tissues and cells lines. It was found that PTK2 was a target gene of miR-520d-5p. The expression of miR-520d-5p was down-regulated in CC tissues, which was negatively correlated with the expression of PTK2. MiR-520d-5p inhibited the proliferation, migration, and invasion of CC cells. In addition, overexpression of miR-520d-5p resulted in apoptosis of CC cells. Finally, we demonstrated that miR-520d-5p inhibited the activation of PI3K/AKT signaling.

CONCLUSION

MiR-520d-5p suppressed the proliferation, invasion, and migration of CC cells via targeting PTK2.

摘要

目的

已有报道称 PTK2 参与肿瘤进展，但其在宫颈癌（CC）中的调节机制仍不清楚。miRNA-520d-5p 被证明可以调节许多基因的表达并抑制人类肿瘤的发展。然而，miRNA-520d-5p 调节宫颈癌的功能机制尚不完全清楚。

方法

采用 RT-qPCR 检测 miR-520d-5p 和 PTK2 的表达水平。采用 Western blot 检测蛋白表达水平。采用双荧光素酶报告基因实验研究 miR-520d-5p 和 PTK2 之间的关联。采用 CCK-8 assay 检测细胞增殖。此外，还进行了 Transwell assay 和划痕 assay 以分析细胞侵袭和迁移。采用流式细胞术检测宫颈癌细胞凋亡。

结果

PTK2 在 CC 组织和细胞系中的表达水平升高。研究发现，PTK2 是 miR-520d-5p 的靶基因。CC 组织中 miR-520d-5p 的表达下调，与 PTK2 的表达呈负相关。miR-520d-5p 抑制 CC 细胞的增殖、迁移和侵袭。此外，miR-520d-5p 的过表达导致 CC 细胞凋亡。最后，我们证明 miR-520d-5p 通过靶向 PTK2 抑制 PI3K/AKT 信号通路的激活。