Abstract:
Background MicroRNA (miR)-324-3P plays an important role in a variety of malignant tumors, regulation of Wnt/β- catenin signaling pathway may be an important pathway involved in tumor progression, while the specific role of miR-324-3P in cervical cancer remains unclear.
Objective To study the effect of microRNA (miR)-324-3P on the proliferation, invasion and metastasis of cervical cancer cells and its mechanism.
Methods Forty cases of cervical cancer tissues and normal cervical tissues were collected from patients with cervical cancer admitted to our hospital. TargetScan 7.1 prediction software and double luciferase reporter gene test were used to detect the targeting regulation of miR-324-3P on WNT3a gene. Real time fluorescent quantitative PCR (QRT PCR) was used to detect expression of miR-324-3P and WNT3a mRNA in cervical cancer tissues and normal cervical tissues. Pearson was used to analyze the correlation between miR-324-3P and WNT3a expression in cervical cancer tissues. HeLa cells were routinely cultured and divided into control group (NC group), miR-324-3P mimic group and miR-324-3P mimic+oe-WNT3a group. Lip2000 was used to transfect the plasmids of each group, Tetrazolium salt colorimetry (MTT) test was used to detect the cell proliferation ability, Boyden test was used to detect the invasion ability, Transwell test was used to detect the metastasis ability, and Western blot (Western-blot) was used to detect the expression of WNT3a and β-catenin in each group.
Results The relative expression levels of miR-324-3P and WNT3a were (0.79±0.39) and (1.46±0.53), respectively, compared with normal cervical tissues, the expression of miR-324-3P decreased, and the expression of WNT3a increased. Targetscan7.1 prediction software, dual luciferase reporter assay and qRT-PCR showed that miR-324-3P targeted WNT3a, and Pearson analysis showed that there was a negative correlation between them. Compared with the NC group, the proliferation, invasion and metastasis ability of cervical cancer cells in the miR-324-3P mimic group and the miR-324-3P mimic+oe-WNT3a group were reduced. Compared with the miR-324-3P mimic group, the proliferation, invasion and metastasis ability of cervical cancer cells in the miR-324-3P mimic+oe-WNT3a group were increased, and WNT3a attenuated the effect of miR-324-3P on the inhibition proliferation, invasion and metastasis ability of cervical cancer cells. Compared with NC group, the expressions of WNT3a and β-catenin decreased in miR-324-3P mimic group and miR-324-3P mimic+oe-WNT3a group. Compared with miR-324-3P mimic group, the expression of WNT3a and β-catenin protein increased in miR-324-3P mimic+oe-WNT3a group. WNT3a attenuated the inhibitory effect of miR-324-3P on β-catenin.
Conclusion miR-324-3P targets WNT3a expression and inhibits the proliferation, invasion and metastasis of cervical cancer cells.