Roquin1 inhibits the proliferation of breast cancer cells by inducing G1/S cell cycle arrest via selectively destabilizing the mRNAs of cell cycle–promoting genes

Abstract

Background Dysregulation of cell cycle progression is a common feature of human cancer cells; however, its mechanism remains unclear. This study aims to clarify the role and the underlying mechanisms of Roquin1 in cell cycle arrest in breast cancer. Methods Public cancer databases were analyzed to identify the expression pattern of Roquin1 in human breast cancers and its association with patient survival. Quantitative real-time PCR and Western blots were performed to detect the expression of Roquin1 in breast cancer samples and cell lines. Cell counting, MTT assays, flow cytometry, and in vivo analyses were conducted to investigate the effects of Roquin1 on cell proliferation, cell cycle progression and tumor progression. RNA sequencing was applied to identify the differentially expressed genes regulated by Roquin1. RNA immunoprecipitation assay, luciferase reporter assay, mRNA half-life detection, RNA affinity binding assay, and RIP-ChIP were used to explore the molecular mechanisms of Roquin1. Results We showed that Roquin1 expression in breast cancer tissues and cell lines was inhibited, and the reduction in Roquin1 expression was associated with poor overall survival and relapse-free survival of patients with breast cancer. Roquin1 overexpression inhibited cell proliferation and induced G1/S cell cycle arrest without causing significant apoptosis. In contrast, knockdown of Roquin1 promoted cell growth and cycle progression. Moreover, in vivo induction of Roquin1 by adenovirus significantly suppressed breast tumor growth and metastasis. Mechanistically, Roquin1 selectively destabilizes cell cycle–promoting genes, including Cyclin D1, Cyclin E1, cyclin dependent kinase 6 (CDK6) and minichromosome maintenance 2 (MCM2), by targeting the stem–loop structure in the 3′ untranslated region (3’UTR) of mRNAs via its ROQ domain, leading to the downregulation of cell cycle–promoting mRNAs. Conclusions Our findings demonstrated that Roquin1 is a novel breast tumor suppressor and could induce G1/S cell cycle arrest by selectively downregulating the expression of cell cycle–promoting genes, which might be a potential molecular target for breast cancer treatment.

Roquin2 suppresses breast cancer progression by inhibiting tumor angiogenesis via selectively destabilizing proangiogenic factors mRNA

“…Roquin1/2 has been shown to play an important role in suppressing autoinflammation by enhancing the mRNA decay of proinflammatory cytokines 31 . In our previous study, we found that Roquin1 was able to suppress breast tumor progression by inducing cell cycle arrest of breast cancer cells 32 . However, there are very few reports on the role of Roquin2 in tumor progression 26 , 27 .…”

RNA‐binding protein SAMD4A inhibits breast tumor angiogenesis by modulating the balance of angiogenesis program

“…RIP experiment was conducted as previously described 33 . The protein‐RNA complexes were immunoprecipitated by protein A/G beads, and total RNA extracted with TRIzol, followed by detection with RT‐PCR.…”

Arbidol inhibits human esophageal squamous cell carcinoma growth in vitro and in vivo through suppressing ataxia telangiectasia and Rad3-related protein kinase

“…ATR can phosphorylate proteins involved in DNA replication, DNA recombination and repair, and cell cycle regulation in response to DNA damage ( Snedeker et al, 2017 ). Due to its extensive role in DDR, ATR inhibition has great potential in cancer treatment ( Cheng et al, 2017 ; Jin et al, 2021 ; Lu et al, 2020 ). Therefore, suppressing ATR signal transduction by inhibitors may exert antitumor effects against ESCC ( Gorecki et al, 2020 ).…”