BMP9对骨肉瘤细胞侵袭迁移能力的影响及机制

Effects of BMP9 on invasion and migration of osteosarcoma cells and its mechanism

目的探讨骨形态发生蛋白9(BMP9)对骨肉瘤细胞侵袭、迁移能力的影响及机制。方法以骨肉瘤细胞F5M2为研究对象,通过慢病毒感染过表达细胞中BMP9(Ad BMP9组),以空慢病毒载体为Ad GFP组,以不处理的细胞为对照组,通过RT-PCR和Western blot检测感染后细胞中BMP9表达水平,细胞划痕实验检测细胞迁移能力,Transwell小室检测细胞侵袭能力,Western blot检测细胞中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、β-连环蛋白(β-catenin)表达。结果对照组、Ad GFP组、Ad BMP9组细胞中BMP9 mRNA水平依次为(0.38±0.06)、(0.36±0.05)、(0.92±0.04),BMP9蛋白表达水平依次为(0.20±0.02)、(0.21±0.06)、(1.22±0.17),迁移率依次为(99.87±4.36)%、(100.03±8.64)%、(66.58±4.25)%,侵袭细胞数依次为(115.04±4.98)、(114.95±4.85)、(77.68±8.34),MMP-9水平依次为(0.88±0.06)、(0.87±0.08)、(0.35±0.07),MMP-2水平依次为(0.98±0.09)、(0.99±0.06)、(0.32±0.01),β-catenin水平依次为(1.56±0.09)、(1.58±0.05)、(0.22±0.04)。Ad GFP组细胞中BMP9 mRNA和蛋白表达水平、迁移率、侵袭细胞数、MMP-2水平、MMP-9水平、β-catenin水平与对照组比较差异均无统计学意义(P>0.05)。Ad BMP9组细胞中BMP9 mRNA和蛋白表达水平与对照组比较均明显升高,差异均有统计学意义(P<0.05),而迁移率、侵袭细胞数、MMP-2水平、MMP-9水平、β-catenin水平与对照组比较均明显降低,差异均有统计学意义(P<0.05)。结论过表达BMP9能够降低骨肉瘤细胞迁移侵袭能力,其作用机制可能与MMP-2、MMP-9、β-catenin有关。

Objective To explore the effect of BMP9 on the invasion and migration of osteosarcoma cells andits mechanism. Methods Osteosarcoma cell F5M2 was used as the research object, and BMP9 was overexpressed in in-fected cells by lentivirus （AdBMP9 group）. Slow empty virus vector, were taken as the AdGFP group, and untreatedcells were used as the control group. BMP9 expression in infected cells was detected by RT-PCR and Western blot; cellscratch assay was used to detect the ability of cell migration; cell invasion was detected by Transwell chamber; MMP-2,MMP-9 and β-catenin expression in cells were detected by Western blot. Results The expression levels of BMP9mRNA in the control group, AdGFP group and AdBMP9 group were as follows： （0.38±0.06）, （0.36±0.05）, （0.92±0.04）;the expression levels of BMP9 protein were （0.20±0.02）, （0.21±0.06） and （1.22±0.17）, respectively; the migration rateswere as follows： （99.87±4.36）%, （100.03±8.64）% and （66.58±4.25）%; the number of invasive cells were as follows：（115.04±4.98）, （114.95±4.85）, （77.68±8.34）; the expression levels of MMP-9 protein in the control group, AdGFP groupand AdBMP9 group were as follows： （0.88±0.06）, （0.87±0.08）, （0.35±0.07）; the expression levels of MMP-2 proteinwere （0.98±0.09）, （0.99±0.06）, and （0.32±0.01）, respectively; the protein levels of β-catenin were （1.56±0.09）, （1.58±0.05）, and （0.22±0.04）, respectively. There was no significant difference between AdGFP group and the control group inthe expression of BMP9 mRNA and protein, the rate of migration, invasion cell number, MMP-2 protein expression lev-el, MMP-9 protein expression level, β-catenin level （P〉0.05）. The levels of BMP9 mRNA and protein in the AdBMP9group were significantly higher than those in the control group （P〈0.05）, while the rate of migration, invasion cell num-ber, MMP-2 level, MMP-9 level, β-catenin levels were significantly lower than those in the control group （P〈0.05）.Conclusion Overexpression of BMP9 can decrease the migration and invasion of osteosarcoma cells, and the mecha-nism may be related to MMP-2, MMP-9, β-catenin.