Department of Medical Service, Affiliated Hospital of Hebei University of Engineering, Handan, China.
Department of General Surgery, Affiliated Hospital of Hebei University of Engineering, Handan, China.
Cancer Sci. 2022 Jan;113(1):109-119. doi: 10.1111/cas.15113. Epub 2021 Nov 24.
The purpose of this study was to investigate the pathogenesis of colorectal cancer (CRC) and the effects of the long noncoding RNA plasmacytoma variant translocation 1 (PVT1) on CRC progression. Bioinformatics analysis verified PVT1 expression in tumor and normal tissues. Quantitative PCR and western blotting were used to measure mRNA and protein levels, respectively. The MTT, Transwell, colony formation, and in vivo assays were used to assess the effects of PVT1 on proliferation, migration, and invasion by CRC cells. Both PVT1 and microRNA (miR)-152-3p were shown to be colocalized in CRC cells using FISH assay. The target genes of miR-152-3p were predicted and verified by bioinformatics analysis, luciferase assay, and RNA pull-down assay. The ChIP assay revealed that E2F3 binds with the promoter of MAPK8. We found that PVT1 was overexpressed in CRC specimens, and its expression was higher in CRC cells than normal intestinal cells. Overexpression of PVT1 enhanced the proliferation, migration, and invasion of CRC cells, whereas PVT1 knockdown inhibited these processes. MicroRNA-152-3p was a target of PVT1, and E2F3 was a target of miR-152-3p. Rescue experiments confirmed the interaction between miR-152-3p and PVT1 and between miR-152-3p and E2F3. Luciferase and ChIP assay results confirmed that E2F3 modulates the transcriptional activation of MAPK8. Long noncoding RNA PVT1 activated E2F3 signaling by sponging miR-152-3p. The PVT1/miR-152-3p/E2F3/MAPK8 axis promoted CRC progression.
本研究旨在探讨结直肠癌（CRC）的发病机制以及长链非编码 RNA 浆细胞瘤变异易位 1（PVT1）对 CRC 进展的影响。生物信息学分析验证了肿瘤和正常组织中 PVT1 的表达。定量 PCR 和 Western blot 分别用于测量 mRNA 和蛋白水平。MTT、Transwell、集落形成和体内实验用于评估 PVT1 对 CRC 细胞增殖、迁移和侵袭的影响。荧光原位杂交（FISH）检测显示 PVT1 和 microRNA（miR）-152-3p 在 CRC 细胞中存在共定位。通过生物信息学分析、荧光素酶报告基因检测和 RNA 下拉实验预测和验证了 miR-152-3p 的靶基因。染色质免疫沉淀（ChIP）实验显示 E2F3 与 MAPK8 启动子结合。我们发现 PVT1 在 CRC 标本中高表达，且在 CRC 细胞中的表达高于正常肠细胞。过表达 PVT1 增强了 CRC 细胞的增殖、迁移和侵袭能力，而 PVT1 敲低则抑制了这些过程。miR-152-3p 是 PVT1 的靶基因，E2F3 是 miR-152-3p 的靶基因。挽救实验证实了 miR-152-3p 与 PVT1 之间以及 miR-152-3p 与 E2F3 之间的相互作用。荧光素酶和 ChIP 实验结果证实 E2F3 通过海绵吸附 miR-152-3p 调节 MAPK8 的转录激活。长链非编码 RNA PVT1 通过海绵吸附 miR-152-3p 激活 E2F3 信号通路。PVT1/miR-152-3p/E2F3/MAPK8 轴促进 CRC 进展。
