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Archive > Volume 25 Issue 12 > 2018,25(12):1270-1275. DOI:10.3872/j.issn.1007-385X.2018.12.010 Prev Next
Basic Research
Over-expression of miR-488-5p decreases proliferation and migration ability of cervical cancer C33A cells through TEM8
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Abstract:
Objective: To observe the expression of miR-488-5p in cervical cancer tissues and to explore its effect on the proliferation and migration of cervical cancer C33A cells. Methods: 12 pairs of cervical cancer tissues and corresponding paracancer tissues from patients,who underwent total hysterectomy at the Luoyang Central Hospital of Zhengzhou University from March 2017 to September 2017, were collected for this study; and the expression of miR-488-5p was detected by fluorescence quantitative and real-time polymerase chain reaction (qRT-PCR). Lipofectamine 3000 was used to transfect miR-488-5p (experiment group) and miR-NC (control group) into cervical cancer C33A cells. Cell cycle distribution was detected by Flow cytometry. Cell proliferation was assessed by CCK-8 assay and Transwell assay was used to detect cell migration. Bioinformatics software was used to predict the possible target genes of miR-488-5p, and luciferase activity assay was used to verify the binding of miR-488-5p to target genes. The expressions of tumor endothelial marker 8 (TEM8) and downstream EGFR signaling pathway related proteins in two groups were detected by qRT-PCR and Western blotting. Results: The relative expression level of miR-488-5p in cervical cancer tissues (1.33±0.20) was significantly lower than that in paracancer tissues (3.68±0.45) (P<0.01). The relative expression level of miR-488-5p in the experimental group (25.23±3.11)was significantly higher than that in the control group (1.02±0.10) (P<0.01). The percentage of C33A cells at G0/G1 phase in experimental group (53.39±2.48)% was significantly higher than that in control group (39.57±1.21)% (P<0.01). When the culture time extended to 96 h and 120 h, the proliferation ability of C33A cells in experimental group was significantly lower than that in control group (P<0.05), and the number of migrated cells in the experimental group (117.90±18.86) was significantly less than that in the control group (295.10±19.33) (P <0.01). Luciferase activity assay confirmed that miR-488-5p could directly bind with TEM8 and inhibit its expression (P<0.01). The relative expression of TEM8 mRNA in experimental group (0.42±0.06) was significantly lower than that in control group (1.00 ± 0.06) (P<0.01). After transfection with miR-488-5p for 48h, the protein expressions of TEM8, p-EGFR, p-ERK and p-AKT were significantly lower than those in control group (P <0.01). Conclusion: The expression of miR-488-5p in cervical cancer tissues was decreased. Over-expression of miR-488-5p could inhibit the cell cycle progression of cervical cancer cells and reduce the proliferation and migration of cervical cancer cells. The mechanism may be related to the interference of TEM8 gene expression.
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