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检测 microRNA-145-5p 和磷酸丝氨酸转氨酶 1 在结肠癌中的作用。

Examination of the effects of microRNA-145-5p and phosphoserine aminotransferase 1 in colon cancer.

机构信息

Department of Anorectal Surgery, Taizhou First People's Hospital, Taizhou, Zhejiang Province, China.

出版信息

Bioengineered. 2022 May;13(5):12794-12806. doi: 10.1080/21655979.2022.2071010.

Abstract

Previous studies manifested that microRNA-145-5p is pivotal in the development of various cancers. Nevertheless, the potential function of microRNA-145-5p in colorectal cancer remains unclear. This study attempted to investigate the potential role and possible mechanism of microRNA-145-5p in colon cancer. MicroRNA-145-5p and phosphoserine aminotransferase 1 (PSAT1) levels in colon cancer cells were assayed via quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell proliferation and cell cycle status were assessed using Cell Counting Kit-8, colony formation, and flow cytometry. The target binding relationship of microRNA-145-5p and PSAT1 was identified using bioinformatics analysis and dual-luciferase reporter gene assay. The result of qRT-PCR disclosed that microRNA-145-5p was markedly down-regulated and PSAT1 level was up-regulated in colon cancer cell lines. Besides, enforced microRNA-145-5p level repressed proliferation of colon cancer cells, and cells were arrested in G0-G1 phase. Bioinformatics analysis and dual-luciferase reporter genes confirmed that PSAT1 was a downstream target of microRNA-145-5p. Enforced PSAT1 level remarkably modulated cell cycle and fostered cell proliferation. Furthermore, rescue experiments displayed that microRNA-145-5p restrained cell cycle progression and cell proliferation and forced PSAT1 level could partially reverse this process. Taken together, our findings demonstrated that microRNA-145-5p repressed colon cancer cell cycle progression and cell proliferation via targeting PSAT1. Our findings identified microRNA-145-5p as an essential tumor repressor gene in colon cancer and may provide a novel biomarker for colon cancer.

摘要

先前的研究表明 microRNA-145-5p 在多种癌症的发展中起着关键作用。然而,microRNA-145-5p 在结直肠癌中的潜在功能尚不清楚。本研究试图探讨 microRNA-145-5p 在结肠癌中的潜在作用和可能的机制。通过定量逆转录聚合酶链反应(qRT-PCR)检测结肠癌细胞中 microRNA-145-5p 和磷酸丝氨酸氨基转移酶 1(PSAT1)的水平。使用细胞计数试剂盒-8、集落形成和流式细胞术评估细胞增殖和细胞周期状态。通过生物信息学分析和双荧光素酶报告基因检测鉴定 microRNA-145-5p 和 PSAT1 的靶标结合关系。qRT-PCR 的结果表明,结肠癌细胞系中 microRNA-145-5p 明显下调,PSAT1 水平上调。此外,强制表达 microRNA-145-5p 水平抑制结肠癌细胞的增殖,并使细胞停滞在 G0-G1 期。生物信息学分析和双荧光素酶报告基因证实 PSAT1 是 microRNA-145-5p 的下游靶标。强制表达 PSAT1 水平显著调节细胞周期并促进细胞增殖。此外,挽救实验显示,microRNA-145-5p 抑制细胞周期进程和细胞增殖,而强制表达 PSAT1 水平可部分逆转这一过程。总之,我们的研究结果表明,microRNA-145-5p 通过靶向 PSAT1 抑制结肠癌细胞周期进程和细胞增殖。我们的研究结果确定 microRNA-145-5p 是结直肠癌中重要的肿瘤抑制基因,可为结直肠癌提供新的生物标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e55/9275947/daba4c614b14/KBIE_A_2071010_UF0001_OC.jpg

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