IGF2BP1 通过 mA 修饰增强 SIK2 mRNA 的稳定性，从而促进非小细胞肺癌进展。

IGF2BP1 enhances the stability of SIK2 mRNA through mA modification to promote non-small cell lung cancer progression.

机构信息

The Second Department of Thoracic Oncology, Hunan Cancer Hospital, The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, 410013, Hunan Province, PR China.

出版信息

Biochem Biophys Res Commun. 2023 Dec 3;684:149113. doi: 10.1016/j.bbrc.2023.10.045. Epub 2023 Oct 12.

DOI:10.1016/j.bbrc.2023.10.045

PMID:37866243

Abstract

BACKGROUND

Non-small cell lung cancer (NSCLC) is a significant public health concern globally. Evidence suggests that Salt-inducible kinase 2 (SIK2) is differentially expressed across various cancers and is also implicated in cancer progression. Despite this, the precise function of SIK2 in NSCLC is yet to be elucidated and requires further investigation.

METHODS

SIK2 expression was evaluated in both HBEC and NSCLC cells, utilizing quantitative real-time PCR (qRT-PCR) and Western blot (WB) analyses. Furthermore, to identify the influence of SIK2 on cell proliferation, migration, invasion, and apoptosis, a range of techniques were employed. To evaluate N6-methyladenosine (mA) modification levels of total RNA and SIK2 within cells, RNA mA colorimetry and methylated RNA immunoprecipitation (MeRIP) techniques were employed. Additionally, to confirm the interaction between SIK2 and insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1), bioinformatics analysis was executed, and the results were validated through RIP. The stability of SIK2 mRNA was determined using actinomycin D experiment. Furthermore, to validate the in vivo functionality of SIK2, a subcutaneous transplantation tumor model was established in nude mice.

RESULTS

In this study, upregulation of SIK2 in NSCLC cells was observed. Overexpression of SIK2 was found to lead to promotion of cell proliferation, migration, invasion, and suppression of the Hippo/yes-associated protein (YAP) pathway, while inhibiting apoptosis. RIP analysis showed that IGF2BP1 protein interacted with SIK2 mRNA. Knockdown of IGF2BP1 decreased mRNA stability and mA modification levels of SIK2. Additionally, knockdown of IGF2BP1 resulted in inhibition of cell proliferation, migration, invasion, suppression of the Hippo/YAP pathway, and promoting apoptosis. Overexpression of SIK2 overturned the impact of IGF2BP1 on NSCLC cells, which was then confirmed through in vivo experiments.

CONCLUSION

IGF2BP1 stabilized SIK2 mRNA through mA modification to promote NSCLC progression, potentially offering new diagnostic and therapeutic insights for NSCLC.

摘要

背景

非小细胞肺癌（NSCLC）是一个全球性的重大公共卫生问题。有证据表明，盐诱导激酶 2（SIK2）在各种癌症中表达不同，并且与癌症进展有关。尽管如此，SIK2 在 NSCLC 中的确切功能仍有待阐明，需要进一步研究。

方法

通过定量实时 PCR（qRT-PCR）和 Western blot（WB）分析评估 SIK2 在 HBEC 和 NSCLC 细胞中的表达。此外，为了确定 SIK2 对细胞增殖、迁移、侵袭和凋亡的影响，采用了一系列技术。为了评估细胞内总 RNA 和 SIK2 的 N6-甲基腺苷（mA）修饰水平，采用了 RNA mA 比色法和甲基化 RNA 免疫沉淀（MeRIP）技术。此外，为了确认 SIK2 与胰岛素样生长因子 2 mRNA 结合蛋白 1（IGF2BP1）之间的相互作用，进行了生物信息学分析，并通过 RIP 验证了结果。通过放线菌素 D 实验确定了 SIK2 mRNA 的稳定性。此外，为了验证 SIK2 的体内功能，在裸鼠中建立了皮下移植肿瘤模型。

结果

本研究观察到 NSCLC 细胞中 SIK2 的上调。过表达 SIK2 导致细胞增殖、迁移、侵袭的促进，Hippo/yes 相关蛋白（YAP）通路的抑制，以及凋亡的抑制。RIP 分析表明，IGF2BP1 蛋白与 SIK2 mRNA 相互作用。IGF2BP1 的敲低降低了 SIK2 mRNA 的稳定性和 mA 修饰水平。此外，IGF2BP1 的敲低抑制了细胞增殖、迁移、侵袭、Hippo/YAP 通路的抑制以及凋亡的促进。SIK2 的过表达推翻了 IGF2BP1 对 NSCLC 细胞的影响，这一点随后通过体内实验得到了证实。