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Figure 6 | Journal of Translational Medicine

Figure 6

From: Tumor suppressor miR-1 restrains epithelial-mesenchymal transition and metastasis of colorectal carcinoma via the MAPK and PI3K/AKT pathway

Figure 6

LASP1 was a direct target of miR-1 in CRC cells. (A) Diagram of LASP1 3’UTR containing 2 putative conserved target sites for miR-1, which were identified using the TargetScan database. (B) Results of luciferase reporter assays in 293 T cells, with cotransfection of wt or mt 3’UTR and miR mimic and inhibitor, as indicated. (C) Expression of miR-1 and LASP1 mRNA were detected in CRC tissues by qRT-PCR analysis. A statistically significant inverse correlation between miR-1 and LASP1 mRNA was observed in CRC specimens. (D) LASP1 protein expression in SW480, SW620, and HCT116 cells 48 hours after transfection with miR-1 or anti-miR-1 was detected by Western blot analysis. The immunosignal was quantified using densitometric scanning software, and relative protein abundance was determined by normalization with β-actin. Each bar represents the mean ± SD. The results were reproduced in 3 independent experiments. The asterisk (*) indicates P < 0.05. (E) SW480 cells transfected with miR-1 mimic and/or LASP1 cDNA were used to determine the role of LASP1 in miR-1–mediated biological behaviors. Expression of LASP1 protein was detected by Western blot analysis. (F) Cell proliferation was determined by CCK-8 assay. (G) Results of transwell assay, which was carried out to evaluate the effect of cell migration after transfection are shown.

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