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目的探讨分化抑制因子1(inhibitor of differentiation factor 1,Id1)对人骨肉瘤细胞MG63恶性逆转向成骨分化的影响。方法用重组腺病毒调控MG63细胞中Id1过表达后,RT-PCR及Western blot检测验证Id1的表达情况,CCK-8检测细胞增殖能力,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,流式细胞术检测细胞周期,ALP读数法检测成骨分化早期指标ALP的活力。结果 Ad Id1重组腺病毒可以使MG63细胞中的Id1表达量增高,Adsi Id1重组腺病毒可以使MG63细胞中的Id1表达量降低(P<0.05);Id1过表达后,MG63细胞的增殖能力、迁移能力、侵袭能力等恶性生物学行为均增强,G1期细胞比例降低,Id1表达量下调后则逆转MG63细胞的恶性生物学行为,G1期细胞比例增高(P<0.05);Id1表达量下调后,正常成骨分化的早期指标碱性磷酸酶(ALP)表达量增高(P<0.05)。结论抑制Id1基因表达可以促使骨肉瘤的恶性生物学行为逆转并诱导骨肉瘤细胞发生成骨分化。
Objective To investigate the effect of Id1 on malignant reverse osteogenic differentiation of human osteosarcoma MG63 cells. Methods Id1 was detected by RT-PCR and Western blot after the Id1 was overexpressed in MG63 cells by recombinant adenovirus. Cell proliferation was assessed by CCK-8. Migration assay was used to detect cell migration. Transwell assay was used to detect cell invasion, Flow cytometry was used to detect the cell cycle. ALP assay was used to detect the activity of ALP, an early indicator of osteogenic differentiation. Results Ad Id1 recombinant adenovirus could increase the expression of Id1 in MG63 cells. The adenovirus Adsi Id1 could reduce the expression of Id1 in MG63 cells (P <0.05). After Id1 overexpression, the proliferation and migration of MG63 cells (P <0.05). After the expression of Id1 was down-regulated, the malignant biological behavior of MG63 cells was reversed and the percentage of cells in G1 phase was increased (P <0.05) Alkaline phosphatase (ALP), an early indicator of normal osteogenic differentiation, increased (P <0.05). Conclusion Inhibition of Id1 gene expression can reverse the malignant biological behavior of osteosarcoma and induce osteosarcoma cell differentiation.