RNAi沉默ADM表达对人骨肉瘤细胞中β-catenin的作用及对成骨分化的影响.

Objective To observe the effect of silencing the expression of ADM using RNA interfering technique on the expression of β-catenin and terminal differentiation of osteosarcoma cells. Methods After the intervention of 0h, 48h and 72h by ADM siRNA, we observed the change of distribution of β-catenin in F5M2 using immunocytochemistry staining. The expression levels of total and phosphorylated β-catenin and GSK3β were detected by Western blot after the intervention by ADM siRNA. The F5M2 cells treated with ADM siRNA were subjected to HE staining, alkaline phosphatase (ALP) assay and immunocytochemistry staining to investigate the biological effects of ADM siRNA on the morphology and terminal differentiation of F5M2 cells. Results After the intervention of 0h, 48h and 72h by ADM siRNA, the distribution of β-catenin was transferred from the cytoplasm to the nucleus. ADM siRNA downregulated the expression level of P-β-catenin and upregulated P-GSK3β detected by Western blot. HE staining revealed that the configuration of F5M2 had undergone restorational changes similar to those of normal cells after ADM siRNA treatment. ALP assay and immunocytochemistry staining showed that the expression of the earlier and later molecular biomarkers of terminal differentiation, including ALP and osteocalcin were strongly positive. Conclusion Silencing the expression of ADM can activate β-catenin to transfer to the nucleus from the cytoplasm, and induce osteosarcoma cells to make terminal differentiation through activating β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]