Li Fusheng, Xu Jia, Zhu Yue
Department of Orthopaedics, The First Affiliated Hospital of China Medical University, 155 Nan Jing Bei Street, Shenyang, 110001, People's Republic of China.
Department of Orthopaedics Oncology, The People's Hospital of Liaoning Province, Shenyang, 110016, People's Republic of China.
Discov Oncol. 2024 May 18;15(1):175. doi: 10.1007/s12672-024-01038-5.
MicroRNAs play an important role in the proliferation, invasion, and metastasis of malignancy. In previous studies (detailed in our previous paper), the expression of miR-6839-5p was significantly increased in SW1353 cells after I seed 6 Gy irradiation, which indicated miR-6839-5p may play a tumor suppression function in chondrosarcoma cells. This study aimed to identify the effects of miR-6839-5p on the human chondrosarcoma cells, and investigate the potential target genes of miR-6839-5p. Firstly, chondrosarcoma cells (SW1353 and CAL78) were transfected with hsa-miR-6839-5p specific mimic. Secondly, Cell viability assay (MTT assay), Colony formation assay, Wound healing assay, Transwell assay, TUNEL staining and Western blotting experiments were performed, and the results proved miR-6839-5p can inhibit chondrosarcoma cells proliferation, migration and invasion. Meanwhile, miR-6839-5p significantly down-regulated apoptosis facilitator Bcl-2 expression, and promoted apoptosis of chondrosarcoma cells. It is reasonable to speculate miR-6839-5p might downregulate Bcl-2 expression to induce apoptosis in SW1353 human chondrosarcoma cells. Lastly, RNA extraction and bioinformatic analysis was performed on SW1353 cells transfected with hsa-miR-6839-5p specific mimic to investigate the potential target genes of miR-6839-5p. A total of 253 differentially expressed mRNA genes (105 up-regulated genes and 148 down-regulated genes) were found, and 23 differentially expressed downregulated genes were identified. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to validate the results, which demonstrated the expression of BST2, VEGFA, FPR3 and PPARA was significantly downregulated by miR-6839-5p mimic. Furthermore, miR-6839-5p inhibitor can restore or partially restore the expression value of the above four genes. The analysis results of miRNA target gene prediction database indicated VEGFA was the most likely direct target gene of miR-6839-5p.
微小RNA在恶性肿瘤的增殖、侵袭和转移中发挥着重要作用。在先前的研究中（详见我们之前的论文），6 Gy照射后SW1353细胞中miR - 6839 - 5p的表达显著增加，这表明miR - 6839 - 5p可能在软骨肉瘤细胞中发挥肿瘤抑制功能。本研究旨在确定miR - 6839 - 5p对人软骨肉瘤细胞的影响，并研究miR - 6839 - 5p的潜在靶基因。首先，用hsa - miR - 6839 - 5p特异性模拟物转染软骨肉瘤细胞（SW1353和CAL78）。其次，进行细胞活力测定（MTT法）、集落形成测定、伤口愈合测定、Transwell测定、TUNEL染色和蛋白质印迹实验，结果证明miR - 6839 - 5p可抑制软骨肉瘤细胞的增殖、迁移和侵袭。同时，miR - 6839 - 5p显著下调凋亡促进因子Bcl - 2的表达，并促进软骨肉瘤细胞凋亡。有理由推测miR - 6839 - 5p可能下调Bcl - 2表达以诱导SW1353人软骨肉瘤细胞凋亡。最后，对用hsa - miR - 6839 - 5p特异性模拟物转染的SW1353细胞进行RNA提取和生物信息学分析，以研究miR - 6839 - 5p的潜在靶基因。共发现253个差异表达的mRNA基因（105个上调基因和148个下调基因），并鉴定出23个差异表达的下调基因。进行定量实时聚合酶链反应（qRT - PCR）以验证结果，结果表明miR - 6839 - 5p模拟物显著下调了BST2、VEGFA、FPR3和PPARA的表达。此外，miR - 6839 - 5p抑制剂可恢复或部分恢复上述四个基因的表达值。miRNA靶基因预测数据库的分析结果表明VEGFA是miR - 6839 - 5p最可能的直接靶基因。
