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目的探讨Eag1对骨肉瘤细胞增殖、迁移与侵袭的影响及其机制。方法将骨肉瘤Saos-2细胞随机分为Eag1沉默组和对照组,分别转染Eag1 siRNA及scramble。分别采用荧光定量PCR法和Western blotting法检测两组Eag1 mRNA和蛋白相对表达量;MTT法检测两组培养24、48、72、96及120 h的细胞增殖能力(以OD值表示),细胞划痕试验检测两组细胞迁移能力(以划痕愈合率表示),Transwell侵袭试验检测两组细胞侵袭能力(以进入膜下的细胞数量表示);采用Western blotting法检测两组STAT3蛋白相对表达量。结果 Eag1沉默组与对照组Eag1 mRNA相对表达量分别为0.50±0.04、1.00±0.12,Eag1蛋白相对表达量分别为0.30±0.06、1.00±0.10,两组比较P均<0.05。Eag1沉默组接种24、48、72、96及120 h的OD值均低于对照组(P<0.05或<0.01)。Eag1沉默组和对照组划痕愈合率分别为42.34%±4.22%、95.12%±2.32%,进入膜下的细胞数量分别为(352±32)、(642±34)个,STAT3蛋白相对表达量分别为0.79±0.15、1.64±0.23,两组比较P<0.05或<0.01。结论 Eag1沉默可抑制骨肉瘤细胞的增殖、迁移及侵袭;下调STAT3蛋白表达可能是其作用机制。
Objective To investigate the effect and mechanism of Eag1 on proliferation, migration and invasion of osteosarcoma cells. Methods Osteosarcoma Saos-2 cells were randomly divided into Eag1 silencing group and control group, transfected with Eag1 siRNA and scramble respectively. The relative expression of Eag1 mRNA and protein in the two groups were detected by real-time PCR and Western blotting respectively. The cell proliferation ability (expressed as OD value) at 24, 48, 72, 96 and 120 h in both groups was detected by MTT assay. The migration ability of the two groups (indicated by the healing rate of scratches) was detected by the stain test. The invasion ability of the two groups of cells was measured by Transwell invasion assay (expressed as the number of cells entering the membrane); the relative expression of STAT3 protein was detected by Western blotting. Results The relative expression levels of Eag1 mRNA in Eag1 silencing group and control group were 0.50 ± 0.04 and 1.00 ± 0.12, respectively. The relative expression levels of Eag1 protein were 0.30 ± 0.06 and 1.00 ± 0.10 respectively, P <0.05 in both groups. The OD values of Eag1 silencing group at 24, 48, 72, 96 and 120 h after inoculation were lower than those in control group (P <0.05 or <0.01). The wound healing rates of Eag1 silencing group and control group were 42.34% ± 4.22% and 95.12% ± 2.32%, respectively. The number of cells entering the membrane were (352 ± 32) and (642 ± 34) respectively, and the relative expression of STAT3 protein Respectively, 0.79 ± 0.15 and 1.64 ± 0.23, P <0.05 or <0.01 respectively. Conclusion Silencing of Eag1 can inhibit the proliferation, migration and invasion of osteosarcoma cells. Down-regulation of STAT3 protein may be the mechanism of its effect.