Figures & data
Figure 1 Elevated STAT3 levels were accompanied by increased miR-572 and decreased MOAP-1 levels in human primary colorectal carcinoma (CRC) specimens and cell lines.
Notes: (A) The correlation between STAT3 and MOAP-1 mRNA levels was determined by qRT-PCR in 40 cases of frozen CRC tissues (P=0.0005). (B) Levels of miR-572 were inversely related to MOAP-1 protein in human CRC cell lines LS174T, SW620, HT29, LOVO, HCT116, and SW480. Relative miR-572 levels and MOAP-1 protein levels were determined by qRT-PCR and Western blot, respectively.
Figure 2 STAT3 upregulated miR-572 expression in colorectal carcinoma cell lines.
Notes: (A) The levels of miR-572 expression in LS174T cells transfected with a constitutively activated mutation of STAT3 (STAT3-CA) or empty vector (control). (B) The levels of miR-572 expression in SW480 cells transfected with siSTAT3 and siControl. (C) The levels of miR-572 expression in serum-starved LS174T cells treated with EGF (50 ng/mL) or without EGF (control). (D) The levels of miR-572 expression in SW480 cells treated with WP1066 (10 μM) or without WP1066 (control). The levels of miR-572 were determined by qRT-PCR after transfection with STAT3 or siSTAT3 for 24 h, or after treatment with EGF or WP1066 for 30 min. The protein levels of STAT3 (A and B), or STAT3 and p-STAT3 (C and D) are shown at the top center of corresponding figures. *P<0.05; **P<0.01.
Figure 3 miR-572 promoted colorectal carcinoma cell growth, migration, and invasion.
Notes: The proliferation of LS174T cells (A) transfected with miR-572 mimic (20 nM) or with control and SW480 cells (B) transfected with miR-572 inhibitor (multiplicity of infection =5) or with control was analyzed by MTT assay. LS174T cells (C) transfected with miR-572 mimic (20 nM) or with control and SW480 cells (D) transfected with miR-572 inhibitor (multiplicity of infection =5) or with control were subjected to transwell migration or invasion assays. *P<0.05; **P<0.01.
Figure 4 miR-572 downregulated MOAP-1 protein expression.
Notes: The protein levels of MOAP-1 were determined by Western blot after transfection with miR-572 mimic (20 nM) or control in LS174T (A) and SW620 (B) cells, or after transfection with miR-572 inhibitor (multiplicity of infection =5) or control in HCT116 (C) and SW480 (D) cells. β-actin was used as loading control. Scanning densitometry of immunoblotting for each panel was measured (lower). *P<0.05; **P<0.01.
Figure 5 miR-572 inhibited MOAP-1 expression.
Notes: (A) Sequence of miR-572 and the predicted miR-572-binding site at MOAP-1 3′UTR. Dual-luciferase reporter assay was applied in LS174T (B) and SW620 (C) cells after co-transfection with established luciferase reporter vectors (including wild-type [WT] or mutated [MUT] MOAP-1 3′UTR) and miR-572 or miR-control. *P<0.05.
Figure 6 STAT3 induced colorectal carcinoma progression via an miR-572-MOAP-1 pathway.
Notes: After transfection with STAT3 siRNA for 24 h, SW480 cells were transfected with miR-572 mimic or MOAP-1 siRNA for 24 h. The relative mRNA levels of STAT3, miR-572, and MOAP-1 were determined by qRT-PCR (A). SW480 cells transfected with STAT3 siRNA or siRNA control alone or in combination with miR-572 mimic or MOAP-1 siRNA were analyzed by MTT assay (B), transwell migration (C) and invasion (D) assays. *P<0.05. **P<0.01.
