Table 1.
miR-193a and WT1 expression levels and the clinical parameters of 25 breast cancer specimens.
Fig 1.
miR-193a expression levels in breast cancer.
(A) qRT-PCR analysis of relative miR-193a expression levels in 25 pairs of breast carcinoma tissues (Tumor) and adjacent non-cancerous tissues (Normal). (B) miR-193a levels in advanced breast cancer (stage III) tissues were lower than that in early breast cancer (stage I–II) tissues. (C) qRT-PCR analysis of relative miR-193a expression levels in the MCF-10A cell line and seven breast cancer cell lines. *p < 0.05.
Fig 2.
The role of miR-193a in regulating breast cancer cell migration, invasion, and proliferation.
(A) Migration assay. (B) Invasion assay. (C) Wound healing assay. (D) Proliferation assay. (E) Colony formation assay. The mean was derived from the cell counts of five fields; each experiment was repeated three times. Representative images of migrated, invaded, or proliferative cells are shown. *p < 0.05.
Fig 3.
Verification of miR-193a targeting of WT1.
(A) Base complementarity between miR-193a and WT1 mRNA. (B) Luciferase reporter assay of HEK293T cells cotransfected with wild-type (WT1 CDS) or mutant (WT1 CDS Mut) pMIR-WT1 CDS and miR-193a mimics or negative control. (C) Western blot analysis of WT1 protein expression in empty vector-negative control (pLVX-NC) cells, or cells overexpressing miR-193a (pLVX-miR-193a). Results are representative of three independent experiments. (D) Immunofluorescence staining for WT1 protein in cells with empty vector-negative control (pLVX-NC) or pLVX-miR-193a. Scale bar = 100 μm. (E) qRT-PCR analysis of relative expression levels of WT1 in 25 pairs of breast carcinoma tissues (Tumor) and adjacent non-cancerous tissues (Normal). (F) WT1 was negatively correlated with miR-193a at mRNA level (n = 25). *p < 0.05.
Fig 4.
(A) WT1 protein expression in MDA-MB-231 and BT549 cells transfected with MSCV-NC, pLVX-miR-193a, MSCV-WT1, or MSCV-WT1 and pLVX-miR-193a. (B) Migration assay, (C) invasion assay, (D) wound healing assay, and (E) colony formation assay of miR-193a–WT1 interaction in regulating breast cancer cell migration, invasion, and proliferation. The mean was derived from the cell counts of five fields; each experiment was repeated three times. Representative images of proliferative cells are shown. *p < 0.05.