Yang Xiancong, Wang Yaoyao, Rong Simin, An Jiayue, Lan Xiaoxu, Yin Baohui, Sun Yunxiao, Wang Pingyu, Tan Boyu, Xuan Ye, Xie Shuyang, Su Zhenguo, Li Youjie
Department of Clinical Laboratory, The Second Medical College of Binzhou Medical University, Yantai, China.
Department of Biochemistry and Molecular Biology, Binzhou Medical University, Yantai, China.
Front Cell Dev Biol. 2023 Jun 12;11:1173491. doi: 10.3389/fcell.2023.1173491. eCollection 2023.
Acute myeloid leukemia (AML) is a malignant proliferative disease affecting the bone marrow hematopoietic system and has a poor long-term outcome. Exploring genes that affect the malignant proliferation of AML cells can facilitate the accurate diagnosis and treatment of AML. Studies have confirmed that circular RNA (circRNA) is positively correlated with its linear gene expression. Therefore, by exploring the effect of SH3BGRL3 on the malignant proliferation of leukemia, we further studied the role of circRNA produced by its exon cyclization in the occurrence and development of tumors. Genes with protein-coding function obtained from the TCGA database. we detected the expression of SH3BGRL3 and circRNA_0010984 by real-time quantitative polymerase chain reaction (qRT-PCR). We synthesized plasmid vectors and carried out cell experiments, including cell proliferation, cell cycle and cell differentiation by cell transfection. We also studied the transfection plasmid vector (PLVX-SHRNA2-PURO) combined with a drug (daunorubicin) to observe the therapeutic effect. The miR-375 binding site of circRNA_0010984 was queried using the circinteractome databases, and the relationship was validated by RNA immunoprecipitation and Dual-luciferase reporter assay. Finally, a protein-protein interaction network was constructed with a STRING database. GO and KEGG functional enrichment identified mRNA-related functions and signaling pathways regulated by miR-375. We identified the related gene SH3BGRL3 in AML and explored the circRNA_0010984 produced by its cyclization. It has a certain effect on the disease progression. In addition, we verified the function of circRNA_0010984. We found that circSH3BGRL3 knockdown specifically inhibited the proliferation of AML cell lines and blocked the cell cycle. We then discussed the related molecular biological mechanisms. CircSH3BGRL3 acts as an endogenous sponge for miR-375 to isolate miR-375 and inhibits its activity, increases the expression of its target YAP1, and ultimately activates the Hippo signaling pathway involved in malignant tumor proliferation. We found that SH3BGRL3 and circRNA_0010984 are important to AML. circRNA_0010984 was significantly up-regulated in AML and promoted cell proliferation by regulating miR-375 through molecular sponge action.
急性髓系白血病（AML）是一种影响骨髓造血系统的恶性增殖性疾病，长期预后较差。探索影响AML细胞恶性增殖的基因有助于AML的准确诊断和治疗。研究证实，环状RNA（circRNA）与其线性基因表达呈正相关。因此，通过探索SH3BGRL3对白血病恶性增殖的影响，我们进一步研究了其外显子环化产生的circRNA在肿瘤发生发展中的作用。从TCGA数据库中获取具有蛋白质编码功能的基因。我们通过实时定量聚合酶链反应（qRT-PCR）检测SH3BGRL3和circRNA_0010984的表达。我们合成了质粒载体并进行了细胞实验，包括通过细胞转染进行细胞增殖、细胞周期和细胞分化实验。我们还研究了转染质粒载体（PLVX-SHRNA2-PURO）与药物（柔红霉素）联合使用以观察治疗效果。使用circinteractome数据库查询circRNA_0010984的miR-375结合位点，并通过RNA免疫沉淀和双荧光素酶报告基因检测验证这种关系。最后，用STRING数据库构建蛋白质-蛋白质相互作用网络。GO和KEGG功能富集分析确定了受miR-375调控的mRNA相关功能和信号通路。我们在AML中鉴定了相关基因SH3BGRL3，并探索了其环化产生的circRNA_0010984。它对疾病进展有一定影响。此外，我们验证了circRNA_0010984的功能。我们发现circSH3BGRL3敲低特异性抑制AML细胞系的增殖并阻断细胞周期。然后我们讨论了相关的分子生物学机制。CircSH3BGRL3作为miR-375的内源性海绵，隔离miR-375并抑制其活性，增加其靶标YAP1的表达，最终激活参与恶性肿瘤增殖的Hippo信号通路。我们发现SH3BGRL3和circRNA_0010984对AML很重要。circRNA_0010984在AML中显著上调，并通过分子海绵作用调节miR-375促进细胞增殖。
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