Sun Rongxin, Muheremu Aikeremujiang, Hu Yihe
Department of Orthopedics, Xiangya Hospital, CentralSouth University, Kaifu District, Changsha, Hunan 86410008, China,
Department of Orthopedics, Sixth Affiliated Hospital of Xinjiang Medical University, Tianshan District, Urumqi, Xinjiang 86830001, China.
Onco Targets Ther. 2018 Dec 14;11:9091-9099. doi: 10.2147/OTT.S181177. eCollection 2018.
Osteosarcoma is a highly malignant osseous sarcoma with poor prognosis. Previous studies indicated that miRNA-30c may play an important role in the development of osteosarcoma, but its mechanism is not yet clear. The current research was carried out to explore the potential applications of miR-30c in the diagnosis and treatment of osteosarcoma.
Real-time PCR and in situ hybridization were used to test the correlation between miR-30c and the onset of osteosarcoma. In vitro transfection of miR-30 mimic was used to test the effect of miR-30c on the development of osteosarcoma. Cell Counting Kit-8, formation of Petri dish clones, in vivo formation of tumor, flow cytometry tests and Transwell analysis were used to assess the effect of miR-330c on the metastatic potential and invasiveness of osteosarcoma.
Reverse transcriptase-PCR analysis and in situ hybridization tests revealed that the expression of miR-30c was lower in the osteosarcoma tissue than in normal bone tissue (<0.05). Low expression of miR-30c was associated with advanced osteosarcoma staging and low cellular differentiation. Multivariate analysis revealed that lower expression of miR-30c was associated with shorter survival of patients (<0.01). U2OS cell growth was significantly inhibited when transfected with miR-30c mimic. Flow cytometry analysis revealed that overexpression of miR-30c could induce apoptosis of osteosarcoma cells. In vitro Petri dish cloning experiment showed that overexpression of miR-30c reduced the cloning ratio of U2OS cells from 21% to 7%. At the same time, overexpression of miR-30c inhibited the formation of sarcoma in nude mice. Transwell experiments indicated that overexpression of miR-30c could reduce the invasiveness of U2OS cells.
Low expression of miR-30c was associated with high probability of onset and aggressiveness of osteosarcoma and shorter patient survival. Upregulation of miR-30c could downregulate the invasiveness of osteosarcoma. Therefore, miR-30 can be used in the development of future diagnostic and therapeutic techniques.
骨肉瘤是一种预后较差的高度恶性骨肿瘤。既往研究表明,miRNA - 30c可能在骨肉瘤的发生发展中起重要作用,但其机制尚不清楚。本研究旨在探讨miR - 30c在骨肉瘤诊断和治疗中的潜在应用。
采用实时荧光定量PCR和原位杂交技术检测miR - 30c与骨肉瘤发病的相关性。通过体外转染miR - 30模拟物来检测miR - 30c对骨肉瘤发展的影响。使用细胞计数试剂盒 - 8、培养皿克隆形成、体内肿瘤形成、流式细胞术检测和Transwell分析来评估miR - 330c对骨肉瘤转移潜能和侵袭性的影响。
逆转录 - PCR分析和原位杂交检测显示,骨肉瘤组织中miR - 30c的表达低于正常骨组织(<0.05)。miR - 30c低表达与骨肉瘤分期较高和细胞分化程度低相关。多因素分析显示,miR - 30c表达较低与患者生存期较短相关(<0.01)。用miR - 30c模拟物转染U2OS细胞后,其生长受到显著抑制。流式细胞术分析显示,miR - 30c过表达可诱导骨肉瘤细胞凋亡。体外培养皿克隆实验表明,miR - 30c过表达使U2OS细胞的克隆率从21%降至7%。同时,miR - 30c过表达抑制裸鼠肉瘤形成。Transwell实验表明,miR - 30c过表达可降低U2OS细胞的侵袭性。
miR - 30c低表达与骨肉瘤发病概率高、侵袭性强及患者生存期短相关。上调miR - 30c可下调骨肉瘤的侵袭性。因此,miR - 30可用于未来诊断和治疗技术的开发。