miR-494 Inhibits Proliferation and Invasion of Osteosarcoma Cells MG-63 Through Downregulating Sox9 Expression
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摘要:目的
探讨miR-494对人骨肉瘤细胞MG-63增殖与侵袭的影响，并验证Sox9是否为miR-494的靶基因。
方法过表达miR-494后，利用CCK-8、克隆形成实验检测MG-63细胞的增殖，利用Transwell实验检测细胞的侵袭能力。Western blot与荧光定量PCR检测Sox9蛋白与mRNA水平的表达。
结果MG-63细胞转染腺病毒miR-494后，miR-494表达明显升高（t=36.78, P=0.000），Ad-miR-494转染组MG-63细胞增殖（F=1.711, P=0.012）、克隆形成（F=2.742, P=0.019）和侵袭能力（F=1.653, P=0.006）较Ad-GFP组下降。过表达miR-494后，Sox9蛋白（F=5.827, P=0.021）和mRNA表达水平（F=5.827, P=0.021）下降。而Sox9的下调使MG-63细胞增殖（t=27.54, P=0.042）、克隆形成（t=29.64, P= 0.026）和侵袭能力（t=32.48, P=0.016）下降。
结论miR-494通过Sox9抑制骨肉瘤细胞MG-63的增殖与侵袭。
Abstract:ObjectiveTo investigate the impact of miR-494 on the proliferation and invasion of human osteosarcoma cells MG-63 and to confirm whether Sox9 is the target gene of miR-494.
MethodsMG-63 cells were transfected with recombinant adenovirus miR-494(Ad-miR-494), CCK-8 and clone formation experiments were employed to determine the proliferation ability of transfected MG-63 cells. Cells invasion abilities were determined by Transwell assay. Real-time PCR was used to analyze the expression of Sox9 mRNA and protein level to confirmthe adenovirus miR-494 expression in MG-63 cells. The protein expression of Sox9 was analyzed by Western blot.
ResultsThe expression of miR-494 was increased obviously (t=36.78, P=0.000) in MG-63 cells which were transfected with Ad-miR-494. CCK-8, clone formation and Transwell experiment results revealed that the proliferation (F=1.711, P=0.012), clone formation (F=2.742, P=0.019) and invasion (F=1.653, P=0.006) abilities of MG-63 cells were markedly inhibited by the overexpression of miR-494. In addition, we observed the decreased Sox9 protein (F=5.827, P=0.021) and mRNA expression (F=5.827, P=0.021) in MG-63 cells which were transfected with Ad-miR-494. And the proliferation (t=27.54, P=0.042), lone formation (t=29.64, P=0.026) and invasion (t=32.48, P=0.016) abilities of MG-63 cells were markedly inhibited by the decreases of Sox9 expression.
ConclusionMG-63 cells proliferation and invasion abilities are suppressed by miR-494, and this process is potentially achieved via suppressing Sox9 expression.
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Key words:
- miR-494 /
- Osteosarcoma /
- Proliferation /
- Invasion
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图 2 过表达miR-494对骨肉瘤细胞MG-63体外增殖的影响
Figure 2 Effects of miR-494 overexpression on proliferation of osteosarcoma cancer cell line MG-63 in vitro
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图 3 Transwell检测miR-494对MG-63细胞侵袭的影响
Figure 3 Effects of miR-494 on MG-63 cells invasion by Transwell assay
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图 4 miR-494对Sox9蛋白和mRNA水平的影响
Figure 4 Effect of miR-494 on Sox9 protein and mRNA levels
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