Fig. 4
MSI2 exerts the metastatic properties in AML. A Cell migration in EGFP-labeled HEL cells transfected with siR-MSI2 or siR-NC was detected by transwell assay. Representative pictures (left panel) from three independent experiments were observed by fluorescence microscope and statistical analysis diagram (right panel) was illustrated by counting cells from lower chambers, scale bar = 200 μm. B Cell migration and invasion increased in MSI2-OE. HEL cells compared to those in NC. HEL cells. Cells from lower chambers of transwell were counted and stained with crystal violet, scale bar = 200 μm. Representative pictures (left panel) and statistical analysis diagram (right panel) were illustrated. C Agarose gel electrophoresis (left panel) and RIP-PCR (right panel) were used to test the interaction between the MSI2 protein and Snail1 mRNA. *P < 0.05 vs. IgG. D Snail1 mRNA stability curves (left panel) were plotted as qRT-PCR expression with time in NC. HEL and MSI2-OE. HEL cells. Half-life (t1/2) was calculated from the stability curves in statistical analysis diagram (right panel). E Relative levels of MSI2 and MMP2 mRNA were determined in BM aspirates from healthy donors (N = 14) and newly diagnosed AML patients (N = 58). Correlation between MSI2 mRNA expression and MMP2 mRNA expression was assessed (P = 0.01, R = 0.30, N = 72). F AML patients (excluding AML-M3 subtype) were divided into the MMP2 high expression group (≥ 1.563, N = 11) and low expression group (< 1.563, N = 29) according to the optimal cutoff value (1.563) for risk score and the OS of the patients was observed (P = 0.0074). (G) Expression of MSI2 and tumor metastasis-associated proteins in Lenti- MSI2 vector- or Lenti- Control vector-transfecting HEL cells. H NOG mice were intravenously transplanted with 5.0 × 106 NC.HEL or MSI2-OE. HEL cells. Kaplan–Meier survival curve was shown, P = 0.0204, N = 6, log-rank test. I Nodules (red arrow) appeared on the livers from two groups of mice. (J) Liver weight (left panel) and spleen weight (right panel) from two groups of mice were measured. N = 4, t-test. K Relative mean fluorescence intensity (MFI) of CD235a and CD45 in liver (left panel) and bone marrow (right panel) from two groups of mice. L, M Expression of Ki67 in liver (L) and spleen (M) was detected by IHC. Representative pictures (left panel) and statistical analysis diagram (right panel) were illustrated. Data are expressed as mean ± SD (error bars). * P < 0.05, ** P < 0.01 and ***P < 0.001, t-test