SOX5对骨肉瘤细胞迁移和侵袭能力的影响

目的 观察Y染色体上的性别决定区相关高迁移率族盒蛋白-5(sex determining region Y-box protein 5,SOX5)对骨肉瘤细胞MG63迁移和侵袭能力的影响,探讨其在骨肉瘤发病和转移中的作用.方法 本研究为前瞻性研究,通过脂质体介导的基因转染技术将SOX5的三个不同的siRNA(siSOX5-1、siSOX5-2和siSOX5-3)瞬时转染骨肉瘤细胞MG63,转染后48 h检测SOX5的mRNA和蛋白表达,从中挑选出干扰效果最明显的siRNA.进而采用博伊登室实验方法分别对干扰前后的骨肉瘤细胞MG63进行体外细胞迁移和侵袭测定,采用免疫印迹(Western blot)检测迁移和侵袭相关蛋白基质金属蛋白酶(matrix metallproteinase,MMP)-2、MMP-9、Twist1和Snail1)]的表达水平.结果 3条针对人的siRNA(siSOX5-1、siSOX5-2、siSOX5-3)均能够降低SOX5 mRNA及蛋白水平表达,其中siSOX5-3干扰效果最明显.siSOX5-3干扰组的迁移细胞数为(52±5)较对照组的(107±9)显著减少(P<0.05),siSOX5-3干扰组的侵袭细胞数为(27±4)较对照组的(71±2)显著减少(P<0.05).Western blot结果显示siSOX5-3干扰组的侵袭相关蛋白(MMP-2、MMP-9、Twist1和Snail1)的表达均较对照组显著降低(P<0.05).结论SOX5 RNA干扰能显著降低骨肉瘤细胞的迁移和侵袭能力,SOX5能促进骨肉瘤细胞的迁移和侵袭.

Effect of SOX5 on migaration and invasion of osteosarcoma MG63 cells

Objective To observe the effect of RNA interference of sex determining region Y-box protein 5 (SOX5) on migration and invasion of MG63 cells, and reveal its role in the pathogenesis and metastasis of osteosarcoma. Methods This study was a prospective study. MG63 cells were transiently transfected with the siRNAs by using Liposome-mediated gene transfection. The mRNA and protein expression of SOX5 was detected 48 h after transfection. Then the most obvious interference efficacy of siRNA was selected. The migration and invasion of MG63 cells were detected by Boyden chambers. The expression level of MMP-2, MMP-9, Twist1 and Snail1 was detected by Western blotting. Results All the three siRNAs targeting SOX5 (siSOX5-1, siSOX5-2, siSOX5-3) could decrease the SOX5 mRNA and protein levels, but siSOX5-3 was the most effective one among the tested siRNAs. SOX5 knockdown significantly reduced the number of migratory MG63 cells (52 ± 5) as compared with control group (107 ± 9) (P<0.05), and that of invasive MG63 cells (27±4) as compared with control group (71±2) (P<0.05). Low expression of SOX5 reduced the expression level of MMP-2, MMP-9, Twist1 and Snail1 (P<0.05). Conclusion SOX5 RNA interference can significantly reduce the migration and invasion of osteosarcoma cells, and SOX5 can promote the migration and invasion of osteosarcoma cells.