Effect of miR-19b-3p targeting coiled-coil domain containing 6 on melanoma cell proliferation, invasion and tumor volume in mice

Abstract

Objective To investigate the mechanism of microRNA (miRNA, miR) -19b-3p targeted regulation of coiled-coil domain containing 6 (CCDC6) in melanoma cell proliferation, invasion and mouse tumor volume. Methods Fifty-six cases of melanoma tissue and fifty-six healthy people were collected. The relative expression of miR-19b-3p and CCDC6 was detected by real-time quantitative polymerase chain reaction (Real-time PCR). The proliferation of WM35 cells was detected by cell counting kit-8 (CCK-8). The invasion ability of WM35 cells was detected by cell invasion assay. The levels of CCDC6 and p-protein kinase B (Akt) protein were detected by Western blotting. The relationship between miR-19b-3p and CCDC6 was verified by luciferase report analysis. The nude mouse tumor model was established to explore the effect of miR-19b-3p on tumor volume. Results Compared with benign sputum tissue (1.43±0.38), miR-19b-3p levels in melanoma tissues (2.89±0.46) were significantly increased (t=6.823, P<0.05), and miR-19b-3p was significantly associated with stage Ⅲ/Ⅳ (χ2=12.087, P<0.05) and distant metastasis (χ2=12.600, P<0.05). Compared with the blank control group [miR-19b-3p: 1.16±0.28; Cell invasion: (26.35±4.73)/HF] and the miR control group [miR-19b-3p: 1.22±0.30; Cell invasion: (28.16±4.80)/HF], the miR-19b-3p expression level and cell invasion ability of miR-19b-3p mimetic group [miR-19b-3p: 4.06±0.28, t=3.977, t=4.003, P<0.05; Cell invasion: (49.64±5.11)/HF, t=3.981, t=4.500, P<0.05] were significantly increased (P<0.05), while the miR-19b-3p expression level and cell invasion ability were significantly decreased in the miR-19b-3p inhibitor group [miR-19b-3p: 0.41±0.08, t=3.771, t=3.579, P<0.05; Cell invasion: (14.47±0.96)/HF, t=3.669, t=4.004, P<0.05], P<0.05. The results of CCK-8 assay showed that the proliferation of miR-19b-3p mimetic cells was significantly increased, while the proliferation was significantly decreased in miR-19b-3p inhibitor group at 24, 48 and 72 h (F=4.026, 4.609, 7.446, P<0.05). Compared with the miR control group [volume: (467.34±36.75) mm3; CCDC6: 1.00±0.12; miR-19b-3p: 2.17±0.42], the volume of melanoma tissue in the miR-19b-3p mimic group [(1 058.66±66.17) mm3] was significantly increased, the expression of CCDC6 was significantly decreased (0.42±0.08), and the level of miR-19b-3p was significantly increased (4.01±0.66), t=22.836, 8.129, 10.663, P<0.05. Conclusion MiR-19b-3p can promote the invasion and proliferation of melanoma cells, and its mechanism is closely related to the regulation of downstream CCDC6 and p-Akt. Key words: Melanoma; MicroRNA; Coiled-coil domain containing 6; Invasion; Proliferation