长链非编码 RNA TPT1-AS1 通过调控 CREB1 抑制 GPX4 抑制卵巢癌中的铁死亡。

Long non-coding RNA TPT1-AS1 inhibits ferroptosis in ovarian cancer by regulating GPX4 via CREB1 regulation.

机构信息

Department of Gynecology, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi, China.

Department of Gynecology, The Second Affiliated Hospital of Shandong First Medical University, Tai'an, Shandong, China.

出版信息

Am J Reprod Immunol. 2024 Aug;92(2):e13864. doi: 10.1111/aji.13864.

DOI:10.1111/aji.13864

PMID:39141012

Abstract

BACKGROUND

Long non-coding RNAs (lncRNAs) play crucial roles in cellular processes, with dysregulation implicated in various diseases, including cancers. The lncRNA TPT1-AS1 (TPT1 Antisense RNA 1) promotes tumor progression in several cancers, including ovarian cancer (OC), but its influence on ferroptosis and interaction with other proteins remains underexplored.

METHODS

In this study, we employed a multi-faceted approach to investigate the functional significance of TPT1-AS1 in OC. We assessed TPT1-AS1 expression in OC specimens and cell lines using RT-qPCR, in situ hybridization (ISH), and fluorescence in situ hybridization (FISH) assays. Functional assays included evaluating the impact of TPT1-AS1 knockdown on OC cell proliferation, migration, invasiveness, and cell cycle progression. Further, we explored and validated the interaction of TPT1-AS1 with other proteins using bioinformatics. Finally, we investigated TPT1-AS1 involvement in erastin-induced ferroptosis using Iron Assay, Malondialdehyde (MDA) assay, and reactive oxygen species (ROS) detection.

RESULTS

Our findings revealed that TPT1-AS1 overexpression in OC correlated with an unfavorable prognosis. TPT1-AS1 knockdown suppressed cell proliferation, migration, and invasiveness. Additionally, TPT1-AS1 inhibited erastin-induced ferroptosis, and in vivo experiments confirmed its oncogenic impact on tumor development. Mechanistically, TPT1-AS1 was found to regulate Glutathione Peroxidase 4 (GPX4) transcription via CREB1 (cAMP response element-binding protein 1) and interact with RNA-binding protein (RBP) KHDRBS3 (KH RNA Binding Domain Containing, Signal Transduction Associated 3) to regulate CREB1.

CONCLUSION

TPT1-AS1 promotes OC progression by inhibiting ferroptosis and upregulating CREB1, forming a regulatory axis with KHDRBS3. These findings highlight the regulatory network involving lncRNAs, RBPs, and transcription factors in cancer progression.

摘要

背景

长链非编码 RNA（lncRNA）在细胞过程中发挥着关键作用，其失调与包括癌症在内的各种疾病有关。lncRNA TPT1-AS1（TPT1 反义 RNA 1）在几种癌症中促进肿瘤进展，包括卵巢癌（OC），但其对铁死亡的影响及其与其他蛋白质的相互作用仍未得到充分探索。

方法

在这项研究中，我们采用了多种方法来研究 TPT1-AS1 在 OC 中的功能意义。我们使用 RT-qPCR、原位杂交（ISH）和荧光原位杂交（FISH）检测评估 OC 标本和细胞系中 TPT1-AS1 的表达。功能检测包括评估 TPT1-AS1 敲低对 OC 细胞增殖、迁移、侵袭和细胞周期进程的影响。此外，我们使用生物信息学探索和验证了 TPT1-AS1 与其他蛋白质的相互作用。最后，我们使用铁测定法、丙二醛（MDA）测定法和活性氧（ROS）检测法研究了 TPT1-AS1 参与依拉司琼诱导的铁死亡的情况。

结果

我们的研究结果表明，OC 中 TPT1-AS1 的过表达与不良预后相关。TPT1-AS1 敲低抑制细胞增殖、迁移和侵袭。此外，TPT1-AS1 抑制依拉司琼诱导的铁死亡，体内实验证实其对肿瘤发展具有致癌作用。机制上，发现 TPT1-AS1 通过 CREB1（cAMP 反应元件结合蛋白 1）调节谷胱甘肽过氧化物酶 4（GPX4）转录，并与 RNA 结合蛋白（RBP）KHDRBS3（KH RNA 结合域包含，信号转导相关 3）相互作用以调节 CREB1。