| [Objective] We investigated the effect of the DKK-1 expression in U266 cells by arsenic trioxide and the differenation of hFOB1.19 cells through the Wnt singal pathway. In clinical researches, we retrospectively studied the relationship between various clinical features and the prognosis of 131 patients with multiple myeloma, as well as the DKK-1 levels of 39 patients.[Methods] MTT method was adopted to observe the inhibition effect that arsenic trioxide had on the proliferation of U266 and hFOB1.19 cells at the dose level of 1-125μmol/L for 1-3 days. Both the DKK-1 mRNA levels in U266 cells and theβ-catenin mRNA levels in hFOB1.19 cells were detected by RT-PCR. ELISA was used to detect the expression of DKK-1 protein, while the expression ofβ-catenin was measured by Western blotting. The alkaline phosphatase expression was assessed by alkaline phosphatase staining and the evaluation of relative activity of alkaline phosphatase in hFOB1.19 cells. In clinical researches, we retrospectively analyzed 131 patients with multiple myeloma,of whom 39 patients'DKK-1 levels were detected.[Results]1. MTT assay showed that U266 cells proliferation was inhibited in different degrees by As2O3 with the concentration from 1 to 125umol/L. The inhitory rate increased along with the arising concentration and duration of As2O3. hFOB 1.19 cells were cultured with different concentrations of As2O3. The proliferation of cells was not significantly inhibited within 24 hours when the concentration was from 0.1 to lumol/L, but was promoted after 24 hours or 48h. However, when the concentration of As2O3 raised to lOumol/L, the proliferation of hFOB 1.19 cells was inhibited in different degrees.2. The DKK-1 mRNA could expressed in U266 cells. DKK-1 mRNA level was negatively correlated with the concentration of As2O3. The expression of DKK-1 protein in the cell culture could not be inhibited by the treatment of 1-2umol/L As2O3. 3. The expression of ALP in hFOB1.19 cells increased after the treatment of 1-2umol/L As2O3. Moreover, the expression ofβ-catenin mRNA was positively correlated with the concentration of As2O3.4. After analyzing 131 clinical cases, we found that some factors, including age, the proportion of bone marrow plasma cells, Hb, albumin,β2-MG, ISS stage and DS stage, were significantly correlated with the survival time. Meanwhile, it was confirmed that the serum DKK-1 level, which was much higher than the control group, was closely related to the ratio of plasma cells in bone marrow.[Conclusion]1. In vitro, As2O3 could inhibit the proliferation of multiple myeloma cell line (U266) and could promote the proliferation of osteoblast hFOB 1.19 within the clinical serum concentration range (0.1-1umol/L).2. DKK-1 protein could express in U266 cells. As2O3 can inhibited the expression of DKK-1 mRNA, but not the expression of protein.3. Since As2O3 could promote the expression ofβ-catenin and ALP protein in osteoblast hFOB 1.19 cells, it can be concluded that the Wnt signaling pathway might, at least in part, play a role in the treatment of As2O3.4. In studies performed on the clinical casese, we showed that the serum DKK-1 level, which was significantly higher than the control group, was closely related to the level of DKK-1.
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