Yao Zhilu, Liu Ning, Lin Hui, Zhou Yingqun
Department of Gastroenterology, Jingan District Zhabei Central Hospital, Shanghai, 200072, People's Republic of China.
Clinical Medical College of Shanghai Tenth People's Hospital, Nanjing Medical University, Nanjing, Jiangsu, 211166, People's Republic of China.
J Hepatocell Carcinoma. 2023 Jul 21;10:1169-1180. doi: 10.2147/JHC.S408542. eCollection 2023.
Hepatocellular carcinoma (HCC) is a severe global health problem, causing many deaths of patients all over the world. Serine and arginine-rich splicing factor 1 (SRSF1) functions as an important oncogenic role in tumorigenesis and progression in HCC. Therefore, therapies targeting SRSF1 may provide promising therapeutic approaches. MiRNAs are virtually involved at the post-transcriptional level and bind to 3' untranslated region (3'-UTR) of their target messenger RNA (mRNA) to suppress expression.
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the expression of SRSF1 and miR-1183 in HCC cell lines. CCK8 assay, colony formation assay and wound healing assay were used to detect the function of miR-1183 in HCC cell lines in vitro. Luciferase reporter assay and Western blot were applied to detect the regulation of particular molecules. Xenograft tumor assay was used to detect the function of miR-1183 in HCC cell lines in vivo. Immunohistochemistry (IHC) was used to detect the expression of SRSF1 in HCC tissues and Xenograft tumors.
In this study, we identified that miR-1183 was downregulated in HCC cell lines. Functional assays indicated that miR-1183-upregulation cells show weakened proliferation ability and migration ability in vitro and inhibit subcutaneous tumor formation in vivo. With respect to the underlying mechanism, we found that miR-1183 function as a tumor suppressor by specifically binding to SRSF1.
This study is the first to demonstrate that miR-1183 function as an important tumor-suppressing role by binding to the 3'-UTR of SRSF1 mRNA and suppressing its protein level in HCC cells in vitro and in vivo. Further, miR-1183 may be a potential target in the prognosis and treatment of HCC.
肝细胞癌（HCC）是一个严重的全球健康问题，导致世界各地许多患者死亡。富含丝氨酸和精氨酸的剪接因子1（SRSF1）在HCC的肿瘤发生和进展中发挥重要的致癌作用。因此，针对SRSF1的治疗可能提供有前景的治疗方法。微小RNA（miRNA）实际上参与转录后水平，并与其靶信使核糖核酸（mRNA）的3'非翻译区（3'-UTR）结合以抑制表达。
采用定量逆转录-聚合酶链反应（qRT-PCR）检测HCC细胞系中SRSF1和miR-1183的表达。采用CCK8法、集落形成试验和伤口愈合试验检测miR-1183在体外HCC细胞系中的功能。应用荧光素酶报告基因试验和蛋白质免疫印迹法检测特定分子的调控。采用异种移植瘤试验检测miR-1183在体内HCC细胞系中的功能。免疫组织化学（IHC）用于检测HCC组织和异种移植瘤中SRSF1的表达。
在本研究中，我们发现miR-1183在HCC细胞系中表达下调。功能试验表明，上调miR-1183的细胞在体外显示出减弱的增殖能力和迁移能力，并在体内抑制皮下肿瘤形成。关于潜在机制，我们发现miR-1183通过特异性结合SRSF1发挥肿瘤抑制作用。
本研究首次证明miR-1183通过在体外和体内与SRSF1 mRNA的3'-UTR结合并抑制其蛋白水平，在HCC细胞中发挥重要的肿瘤抑制作用。此外，miR-1183可能是HCC预后和治疗的潜在靶点。
