Department of Pathology, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University; People's Hospital of Henan University, Zhengzhou, Henan, 450003, People's Republic of China.
Henan International Medical Center, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University; People's Hospital of Henan University, Zhengzhou, Henan, 450003, People's Republic of China.
J Transl Med. 2022 Jan 29;20(1):51. doi: 10.1186/s12967-022-03257-2.
Cervical cancer (CC) is one of the most common gynecological tumors that threatens women's health and lives. Aberrant expression of PIWI-interacting RNA (piRNA) is closely related with a range of cancers and can serve as a tumor promoter or suppressor in proliferation, migration and invasion. In this study, the aim was not only to discover differential expression of piRNA in CC tissue (CC cells) and normal cervical tissue (normal cervical epithelium cells), but also to investigate the biological function and action mechanism of piRNA in CC.
The DESeq2 approach was used to estimate fold change in piRNA between CC tissue and normal cervical tissue. The relative expressions of piRNAs (piRNA-20657, piRNA-20497, piRNA-14633 and piRNA-13350) and RNA m6A methyltransferases/demethylases were detected using RT-qPCR. After intervention with piRNA-14633 and METTL14 expression, the viability of CaSki cells and SiHa cells was detected by CCK8. CC cell proliferation was detected by colony formation assay. Apoptosis rate and cell cycle were detected by flow cytometry. Transwell assay was performed to detect cell migration and invasion. EpiQuik m6A RNA Methylation Quantification Kit was used to evaluate m6A RNA methylation levels. Expression of methyltransferase-like protein 14 (METTL14), PIWIL-proteins and CYP1B1 were detected by RT-qPCR and western blot. The effect of piRNA-14633 on METTL14 was evaluated by a dual-luciferase reporter assay. The in vivo effects of piRNA-14633 on CC was assessed by nude mice experiments.
piRNA-14633 showed high expression in CC tissues and cells, piRNA-14633 mimic (piRNA-14633 overexpression) promoted viability, proliferation, migration and invasion of CaSki cells and SiHa cells. Besides, piRNA-14633 mimic increased m6A RNA methylation levels and METTL14 mRNA stability. Results of dual luciferase reporter assays indicated that METTL14 was a directed target gene of piRNA-14633. Knockdown of METTL14 with siRNA attenuated proliferation, migration and invasion of CC cells. piRNA-14633 increased CYP1B1 expression, while silencing of METTL14 impaired its expression. The effect of piRNA overexpression on METTL14 expression has concentration-dependent characteristics. Results from in vivo experiment indicated that piRNA-14633 promoted cervical tumor growth.
piRNA-14633 promotes proliferation, migration and invasion of CC cells by METTL14/CYP1B1 signaling axis, highlighting the important role of piRNA-14633 in CC.
宫颈癌（CC）是威胁女性健康和生命的最常见妇科肿瘤之一。PIWI 相互作用 RNA（piRNA）的异常表达与多种癌症密切相关，可作为增殖、迁移和侵袭的肿瘤促进因子或抑制因子。本研究不仅旨在发现 CC 组织（CC 细胞）和正常宫颈组织（正常宫颈上皮细胞）中 piRNA 的差异表达，还旨在研究 piRNA 在 CC 中的生物学功能和作用机制。
使用 DESeq2 方法估计 piRNA 在 CC 组织和正常宫颈组织之间的倍数变化。使用 RT-qPCR 检测 piRNA（piRNA-20657、piRNA-20497、piRNA-14633 和 piRNA-13350）和 RNA m6A 甲基转移酶/去甲基酶的相对表达。用 piRNA-14633 和 METTL14 表达干预后，通过 CCK8 检测 CaSki 细胞和 SiHa 细胞的活力。通过集落形成实验检测 CC 细胞的增殖。通过流式细胞术检测细胞凋亡率和细胞周期。通过 Transwell 测定法检测细胞迁移和侵袭。EpiQuik m6A RNA 甲基化定量试剂盒用于评估 m6A RNA 甲基化水平。使用 RT-qPCR 和 Western blot 检测甲基转移酶样蛋白 14（METTL14）、PIWIL 蛋白和 CYP1B1 的表达。通过双荧光素酶报告基因测定评估 piRNA-14633 对 METTL14 的影响。通过裸鼠实验评估 piRNA-14633 对 CC 的体内影响。
piRNA-14633 在 CC 组织和细胞中高表达，piRNA-14633 模拟物（piRNA-14633 过表达）促进了 CaSki 细胞和 SiHa 细胞的活力、增殖、迁移和侵袭。此外，piRNA-14633 模拟物增加了 m6A RNA 甲基化水平和 METTL14 mRNA 的稳定性。双荧光素酶报告基因测定结果表明，METTL14 是 piRNA-14633 的直接靶基因。用 siRNA 敲低 METTL14 可减弱 CC 细胞的增殖、迁移和侵袭。piRNA-14633 增加了 CYP1B1 的表达，而 METTL14 的沉默则削弱了其表达。piRNA 过表达对 METTL14 表达的影响具有浓度依赖性特征。体内实验结果表明，piRNA-14633 促进了宫颈肿瘤的生长。
piRNA-14633 通过 METTL14/CYP1B1 信号轴促进 CC 细胞的增殖、迁移和侵袭，突出了 piRNA-14633 在 CC 中的重要作用。
