Skip to main content
Fig. 6 | Cancer Cell International

Fig. 6

From: The plasma exosomal miR-1180-3p serves as a novel potential diagnostic marker for cutaneous melanoma

Fig. 6

MiR-1180-3p binds to ST3GAL4 and regulate the malignant phenotype of melanoma cells. A Schematic diagram of predicted binding sites of ST3GAL4 with miR-1180-3p. The binding sites was predicted by RNAhybrid 2.2 and miRWalk as described in Methods. The predicted binding sites were deleted in the sequence (ST3GAL4-3′UTR-Del) based on ST3GAL4-3′UTR-WT luciferase reporter plasmids. B The luciferase activity of ST3GAL4 promoter was inhibited by miR-1180-3p. The luciferase reporter assay was performed in 293T cells transfected with ST3GAL4 3′UTR-WT or ST3GAL4 3′UTR-Del, and treated with miR-1180-3p mimic or control as described in Methods. Data from multiple experiments are expressed as the mean ± SD (n = 4). Significant differences were evaluated using two-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001. C Knockdown of ST3GAL4 in melanoma cells. Stable knockdown of ST3GAL4 in SK-MEL-28 cells was generated by lentiviral infection and subjected to RT-Q-PCR analysis as described in the Methods. Data from multiple experiments are expressed as the mean ± SD (n = 3). Significant differences were evaluated using Student’s t-test, *p < 0.05, **p < 0.01, ***p < 0.001. D Knockdown of ST3GAL4 suppress the proliferation of SK-Mel-28. ST3GAL4-deficient SK-Mel-28 cells were seeded into 96-well plates, and cell viability was examined by CCK-8 kit as described in Methods. Data from multiple experiments are expressed as the means ± SD. Significant differences were evaluated using Two-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001. E, F Inhibition of ST3GAL4 attenuates the invasion ability of SK-Mel-28 cells. Transwell assays were performed as described in Methods. Representative images were taken at indicated hours (E). The number of invasive cells per field was calculated, and the data was presented as the means ± SD (n = 4) of each group (F). The significant difference between cells was evaluated by Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001. G, H The regulatory role of miR-1180-3p on melanoma cells were diminished after knocking down ST3GAL4. ST3GAL4-deficient melanoma cells were treated with miR-1180-3p mimics or inhibitors. Transwell assays were performed in SK-Mel-5 (G) and SK-Mel-28 (H) as described in Methods. The number of invasive cells per field was calculated, and the data was presented as the means ± SD (n = 4) of each group. The significant difference between cells was evaluated by Two-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001

Back to article page