Third Department of Gynecology, Northwest Women's and Children's Hospital, No. 1616, Yanxiang Road, Yanta District, Xi'an, 710061, People's Republic of China.
Second Department of Gynecology, Northwest Women's and Children's Hospital, Xi'an , 710061, China.
J Appl Genet. 2023 Sep;64(3):531-543. doi: 10.1007/s13353-023-00772-w. Epub 2023 Aug 4.
The purpose of this study was to investigate the role of circ_0000119 on CC progression and its molecular mechanism. The expression levels of circ_0000119, miR-433-3p, and p21-activated kinase 2 (PAK2) in CC tissues and cell lines were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was assessed using 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, 5-Ethynyl-2'-deoxyuridine (EdU) assay and colony formation assay. Cell cycle and apoptosis were assessed by flow cytometry. Cell migration and invasive ability were examined by Transwell assays. Downstream binding targets of circ_0000119 were predicted by online bioinformatics tools and confirmed by dual luciferase reporter gene assay, RNA immunoprecipitation (RIP) assay, and RNA pull-down assay. The role of circ_0000119/miR-433-3p/PAK2 axis in regulating the CC process was explored by rescue experiments. A xenograft model was constructed to further determine the effect of circ_0000119 on CC tumor growth in vivo. Immunohistochemistry (IHC) assay was conducted for Ki67 expression. Circ_0000119 was aberrantly upregulated in CC tissues and cell lines. Knockdown of circ_0000119 inhibited CC cell proliferation, cell cycle progress, migration, invasion, and promoted apoptosis of CC cells. MiR-433-3p was a binding target of circ_0000119, and PAK2 was a downstream gene of miR-433-3p. MiR-433-3p inhibition reversed the inhibitory effect of silencing circ_0000119 on CC progression. In addition, PAK2 overexpression reversed the effect of miR-433-3p on CC progression. PAK2 expression was regulated by circ_0000119 and miR-433-3p. Moreover, circ_0000119 knockdown reduced tumor growth of CC in vivo. Circ_0000119 was upregulated in CC, and circ_0000119 knockdown suppressed CC malignant development through the miR-433-3p/PAK2 axis.
本研究旨在探讨环状 RNA（circRNA）0000119 在宫颈癌（CC）进展中的作用及其分子机制。通过实时定量聚合酶链反应（qRT-PCR）检测 CC 组织和细胞系中 circ_0000119、miR-433-3p 和 P21 激活激酶 2（PAK2）的表达水平。使用 3-(4,5)-二甲基噻唑 (-z-y1)-3,5-二苯基四唑溴盐（MTT）法、5-乙炔基-2'-脱氧尿苷（EdU）法和集落形成实验评估细胞增殖。通过流式细胞术评估细胞周期和细胞凋亡。通过 Transwell 实验检测细胞迁移和侵袭能力。通过在线生物信息学工具预测 circ_0000119 的下游结合靶点，并通过双荧光素酶报告基因实验、RNA 免疫沉淀（RIP）实验和 RNA 下拉实验进行验证。通过 rescue 实验探讨 circ_0000119/miR-433-3p/PAK2 轴在调节 CC 进程中的作用。构建异种移植模型进一步确定 circ_0000119 对体内 CC 肿瘤生长的影响。通过免疫组化（IHC）检测 Ki67 表达。circ_0000119 在 CC 组织和细胞系中异常上调。circ_0000119 敲低抑制 CC 细胞增殖、细胞周期进展、迁移、侵袭，并促进 CC 细胞凋亡。miR-433-3p 是 circ_0000119 的结合靶点，PAK2 是 miR-433-3p 的下游基因。miR-433-3p 抑制逆转了沉默 circ_0000119 对 CC 进展的抑制作用。此外，PAK2 过表达逆转了 miR-433-3p 对 CC 进展的影响。PAK2 的表达受 circ_0000119 和 miR-433-3p 的调节。此外，circ_0000119 敲低减少了体内 CC 的肿瘤生长。circ_0000119 在 CC 中上调，circ_0000119 敲低通过 miR-433-3p/PAK2 轴抑制 CC 的恶性发展。
