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. 2019 Feb 19;12:1365–1377. doi: 10.2147/OTT.S192069

Figure 4.

Figure 4

miR-140-5p downregulated PAK1 expression by targeting its 3′UTR region.

Notes: (A) Predictive binding sites between PAK1 3′UTR and miR-140-5p and mutant sites in PAK1-MUT reporter. (B) Dual-luciferase reporter assay was carried out to detect the effect of miR-140-5p overexpression on the luciferase activity of PAK1-WT or PAK1-MUT reporter in Tca8113 cells. (C) Tca8113 and SCC-9 cells were transfected with miR-140-5p mimic or miR-NC, followed by the measurement of miR-140-5p expression via RT-qPCR assay at 48 hours after transfection. (D) Western blot assay of PAK1 protein expression in Tca8113 and SCC-9 cells transfected with miR-140-5p or miR-NC at 48 hours upon transfection. (E) PAK1 protein analysis in six randomly selected tumor tissues and corresponding non-carcinomatous tissues. (F) RT-qPCR analysis of PAK1 mRNA expression in 40 pairs of TSCC tissues and corresponding non-carcinomatous tissues. (G) PAK1 mRNA level detection in NHOK and TSCC cell lines (Tca8113, SCC-2, SCC-4, SCC-9, and Cal27). (H) Correlation analysis between PAK1 mRNA level and miR-140-5p level in 40 cases of TSCC tissues. (I) Western blot assay of PAK1 protein expression in Tca8113 and SCC-9 cells transfected with si-NC, si-MALAT1, si-MALAT1 + anti-miR-NC, or si-MALAT1 + anti-miR-140-5p. *P<0.05; **P<0.01.

Abbreviations: NHOK, normal human oral keratinocyte; RT-qPCR, quantitative reverse transcriptase PCR; si-MALAT1, siRNA targeting MALAT1; si-NC, scrambled control siRNA; TSCC, tongue squamous cell carcinoma.