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前期经基因芯片筛选发现,同源盒基因1(distal-less homeobox 1,DLX1)低表达是导致骨肉瘤形成的关键基因。该研究在此基础上,拟通过过表达DLX1探讨其对骨肉瘤细胞MG63迁移和侵袭等生物学特征的影响,并初步揭示其作用途径。采用含DLX1基因的重组腺病毒Ad DLX1(adenovirus distal-less homeobox 1)和空载腺病毒Ad RFP(adenovirus red fl uorescence protein)分别感染人骨肉瘤细胞MG63,观察细胞荧光表达情况,并用RT-PCR和Western blot验证DLX1表达水平;Transwell实验检测细胞迁移和侵袭能力;DAPI染色和流式细胞术检测细胞凋亡水平;CCK-8检测细胞增殖能力;RT-PCR和Western blot检测Wnt/β-catenin信号通路中β-catenin的表达。结果显示,与对照组(Ad RFP感染组)相比,Ad DLX1能显著增加MG63细胞中DLX1的表达,并导致细胞迁移和侵袭能力下降,但细胞增殖和凋亡情况无明显改变。DLX1过表达还可以上调Wnt/β-catenin信号通路中β-catenin的表达。该研究表明,DLX1可通过Wnt/β-catenin信号通路抑制骨肉瘤细胞MG63的迁移和侵袭。
Preliminary screening by gene chip found that the low expression of homeobox 1 (DLX1) is a key gene that leads to the formation of osteosarcoma. On the basis of this study, we intend to explore the effect of DLX1 on the biological characteristics of MG63 such as migration and invasion of osteosarcoma cells, and to reveal its mechanism of action. The human osteosarcoma cell line MG63 was infected with adenovirus distal-less homeobox 1 containing DLX1 gene and Ad RFP (adenovirus red fl uorescence protein), respectively. Western blot was used to detect the expression of DLX1; Transwell assay was used to detect cell migration and invasion; DAPI staining and flow cytometry were used to detect apoptosis; CCK-8 was used to detect cell proliferation; RT-PCR and Western blot were used to detect Wnt / β-catenin Pathway β-catenin expression. The results showed that, compared with the control group (Ad RFP infection group), Ad DLX1 can significantly increase the expression of DLX1 in MG63 cells, and lead to a decline in cell migration and invasion, but no significant changes in cell proliferation and apoptosis. The overexpression of DLX1 can up-regulate the expression of β-catenin in Wnt / β-catenin signaling pathway. This study shows that DLX1 can inhibit the migration and invasion of osteosarcoma MG63 through the Wnt / β-catenin signaling pathway.