Journal: Journal of translational medicine

Article Title: MicroRNA-143 acts as a tumor suppressor through Musashi-2/DLL1/Notch1 and Musashi-2/Snail1/MMPs axes in acute myeloid leukemia.

doi: 10.1186/s12967-023-04106-6

Figure Lengend Snippet: Fig. 5 MSI2 activity is negatively modulated by miR-143 in vitro (A) Key factors of Notch1 signaling pathway were determined in HEL and HL-60 cells transfected with miR-143 or miR-NC mimic. (B, C) Key factors of Notch1 signaling pathway (B) and tumor metastasis-associated proteins (C) were measured in Lenti- MiR-143 vector- or Lenti-Control vector-transfecting HEL cells. (D) Cell growth of MSI2-OE. HEL and NC. HEL cells transfected with miR-143 or miR-NC mimic was analyzed by CCK-8 assay. (E) Clonogenic capacity of Lenti-MSI2 vector- or Lenti-Control vector-transfecting HEL cells after the transfection with miR-143 or miR-NC mimic. Representative pictures (above) and statistical analysis diagram (below) were illustrated, scale bar = 200 μm. (F) Apoptosis in Lenti-MSI2 vector- or Lenti-Control vector-transfecting HEL cells transfected with miR-143 or miR-NC mimic. (G, H) Expression of MSI2, key factors of Notch1 signaling pathway and cancer stemness-related proteins (G) and tumor metastasis-associated proteins (H) were shown in Lenti-MSI2 vector- or Lenti-Control vector-transfecting HEL cells transfected with miR-143 or miR-NC mimic. Data are expressed as mean ± SD (error bars). * P < 0.05, ** P < 0.01 and ***P < 0.001, t-test

Article Snippet: Then the protein was determined using antibodies against MSI2 (Clone: EP1305Y, Abcam, Cambridge, UK), DLL1 (Cat#202301-AP, Proteintech, Wuhan, CN), HES1 (Clone: D6P2U, Cell Signaling Technology (CST), Beverly, MA, USA), cleaved NOTCH1 (Clone: (D1E11) XP®, CST), NANOG (Cat#A3232, ABclonal, Wuhan, CN), OCT4 (Cat#A7920, ABclonal), SOX2 (Cat#A0561, ABclonal), SNAIL1 (Clone: C15D3, CST), MMP2 (Clone: D4M2N, CST), MMP9 (Clone: (D6O3H) XP®, CST), β-actin (Cat#AF7018, Effinity) and GAPDH (Cat#60004-1-1 g, Proteintech), at 1/1,000 dilution for incubation overnight at 4 °C, respectively.

Techniques: Activity Assay, In Vitro, Transfection, Plasmid Preparation, Control, CCK-8 Assay, Expressing

Journal: Journal of translational medicine

Article Title: MicroRNA-143 acts as a tumor suppressor through Musashi-2/DLL1/Notch1 and Musashi-2/Snail1/MMPs axes in acute myeloid leukemia.

doi: 10.1186/s12967-023-04106-6

Figure Lengend Snippet: Fig. 7 Schematic illustrates the mechanism by which miR-143 directly binds to MSI2 and inhibits its efficacy in AML progression. MSI2/DLL1/ Notch1 axis: MSI2 binds directly to DLL1 mRNA and stabilize it at the post-transcriptional level. Then DLL1 agonistic ligands trigger proteolytic cleavage of Notch1 receptors to generate the NICD. NICD/CSL-dependent transcriptional activation of target gene HES1 is upregulated by canonical Notch signaling cascades to maintain stemness of AML cells. MSI2/Snail1/MMPs axis: MSI2 stabilizes Snail1 mRNA and induced transcriptional upregulation of Snail1, which in turn activated both MMP2 and MMP9 to promote AML cell migration. AML, Acute myeloid leukemia; MiR-143, MicroRNA-143; MSI2, Musashi-2; DLL, delta-like canonical Notch ligand; NICD, Notch intracellular domain; MAML, mastermind like protein; CSL, cBF1-suppressor of hairless-LAG1; HES1, hes family bHLH transcription factor 1; Snail1: snail family transcriptional repressor 1; MMP matrix metalloproteinase

Article Snippet: Then the protein was determined using antibodies against MSI2 (Clone: EP1305Y, Abcam, Cambridge, UK), DLL1 (Cat#202301-AP, Proteintech, Wuhan, CN), HES1 (Clone: D6P2U, Cell Signaling Technology (CST), Beverly, MA, USA), cleaved NOTCH1 (Clone: (D1E11) XP®, CST), NANOG (Cat#A3232, ABclonal, Wuhan, CN), OCT4 (Cat#A7920, ABclonal), SOX2 (Cat#A0561, ABclonal), SNAIL1 (Clone: C15D3, CST), MMP2 (Clone: D4M2N, CST), MMP9 (Clone: (D6O3H) XP®, CST), β-actin (Cat#AF7018, Effinity) and GAPDH (Cat#60004-1-1 g, Proteintech), at 1/1,000 dilution for incubation overnight at 4 °C, respectively.

Techniques: Activation Assay, Migration