[微小RNA-300通过负向调控垂体瘤转化基因1抑制骨肉瘤细胞MG63的侵袭和转移]
[MiR-300 inhibits invasion and metastasis of osteosarcoma cell MG63 by negatively regulating PTTG1].
作者信息
Liang D, Wu X, Bai J, Zhang L, Yin C, Zhong W
机构信息
First Department of Joint Surgery, Affiliated Hospital of Weifang Medical College, Weifang Medical University, Weifang 26105, China.
College of Basic Medical Sciences, Weifang Medical University, Weifang 26105, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2021 Feb 25;41(2):285-291. doi: 10.12122/j.issn.1673-4254.2021.02.18.
OBJECTIVE
To investigate the effects of miR-300 and PTTG1 on osteosarcoma invasion and metastasis and explore the molecular mechanism of osteosarcoma invasion and metastasis.
OBJECTIVE
Western blot was used to detect the expression of PTTG1 in human osteoblasts hFOB1.19 and osteosarcoma cell MG63 and to detect the transfection efficiency of cells transfected with PTTG1-knockdown plasmid; Transwell invasion assay and CCK8 assay detected the effects of knockdown of PTTG1 and overexpression of miR-300 on the invasion and proliferation of osteosarcoma cell MG63. On-line prediction and screening of microRNAs (miRNAs) with complementary PTTG1 binding was conducted. qRT-PCR was performed to examine the expression of miR-300 in hFOB1.19 and MG63 cells, and Western blotting was used to detect the expression of PTTG1 in MG63 cells after transfection with a miR- 300 plasmid. Double luciferase assay was used to detect the targeted binding of miR-300 and PTTG, Transwell invasion assay and CCK8 assay were used to detect the effects of overexpression of miR-300 and overexpression of PTTG1 plasmid on invasion and proliferation of osteosarcoma cell line MG63.
OBJECTIVE
PTTG1 was highly expressed in MG63 cells (=0.0002). PTTG1 knockdown significantly inhibited the invasion (=0.0002) and proliferation (=0.0039) of MG63 cells. Based on the results of online prediction of complementary miRNAs to PTTG1 and analysis of the data from NCBI database, miR-300 was determined as the target miRNA in this study. qRT-PCR results showed a significantly decreased expression of miR-300 in MG63 cells (=0.0004). Overexpression of MiR-300 in MG63 cells significantly decreased the expression of PTTG1 (=0.0007), and the expressions of miR-300 and PTTG1 were negatively correlated. Dual luciferase assay showed that miR-300 could specifically bind to PTTG1 (=0.001). Overexpression of PTTG1 could significantly reverse the effect of miR-300 overexpression on invasion (=0.0003) and proliferation (=0.0077) of MG63 cells.
OBJECTIVE
Overexpression of miR-300 can inhibit the invasion and metastasis of osteosarcoma cell MG63 by targeting PTTG1.
目的
研究miR - 300和垂体瘤转化基因1（PTTG1）对骨肉瘤侵袭和转移的影响，探讨骨肉瘤侵袭和转移的分子机制。
目的
采用蛋白质免疫印迹法检测人成骨细胞hFOB1.19和骨肉瘤细胞MG63中PTTG1的表达，并检测转染PTTG1基因敲低质粒的细胞的转染效率；采用Transwell侵袭实验和CCK8实验检测PTTG1基因敲低和miR - 300过表达对骨肉瘤细胞MG63侵袭和增殖的影响。对与PTTG1具有互补结合的微小RNA（miRNA）进行在线预测和筛选。采用qRT - PCR检测hFOB1.19和MG63细胞中miR - 300的表达，并用蛋白质免疫印迹法检测转染miR - 300质粒后MG63细胞中PTTG1的表达。采用双荧光素酶报告基因实验检测miR - 300与PTTG1的靶向结合，采用Transwell侵袭实验和CCK8实验检测miR - 300过表达和PTTG1质粒过表达对骨肉瘤细胞系MG63侵袭和增殖的影响。
目的
PTTG1在MG63细胞中高表达（P = 0.0002）。PTTG1基因敲低显著抑制MG63细胞的侵袭（P = 0.0002）和增殖（P = 0.0039）。基于对与PTTG1互补的miRNA的在线预测结果及美国国立生物技术信息中心（NCBI）数据库数据分析，本研究确定miR - 300为靶标miRNA。qRT - PCR结果显示MG63细胞中miR - 300表达显著降低（P = 0.0004）。MG63细胞中miR - 300过表达显著降低PTTG1的表达（P = 0.0007），且miR - 300与PTTG1的表达呈负相关。双荧光素酶报告基因实验表明miR - 300可特异性结合PTTG1（P = 0.001）。PTTG1过表达可显著逆转miR - 300过表达对MG63细胞侵袭（P = 0.0003）和增殖（P = 0.0077）的影响。
目的
miR - 300过表达可通过靶向PTTG1抑制骨肉瘤细胞MG63的侵袭和转移。
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