Abstract
Purpose
To investigate the effects of miR-1246 on proliferation, invasion, and migration in the human (CSCC) cell line SiHa.
Methods
SiHa cells were assigned into three groups: miR-1246 analog; miR-1246 antagonist; and control. The MTT, transwell, and wound healing assays were performed to evaluate the proliferation, invasion, and migration abilities of SiHa cells, respectively. Western blot was carried out to detect protein expression of thrombospondin-2 (THBS2) before and after transfection with miR-1246 analog, antagonist, or control. In addition, a THBS2 3′-UTR-containing dual luciferase plasmid was generated and co-transfected with miR-1246, the inhibitor, or non-specific miRNA, into SiHa cells to observe its effects on THBS2-driven luciferase enzyme activity.
Results
MTT, transwell, and wound healing assays revealed that proliferation, migration, and invasion were all significantly enhanced (P < 0.01) in SiHa cells transfected with miR-1246 analog, but were suppressed in those transfected with the miR-1246 antagonist. Western blot data showed that miR-1246 analog-transfected SiHa cells had significantly decreased THBS2 expression when compared with control-transfected cells (gray value = 6.28 ± 10.22 vs. 9.58 ± 17.58; P = 0.013) while those transfected with the miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90 ± 19.81; P = 0.037). Moreover, SiHa cells co-transfected with miR-1246 and the THBS2 3′-UTR-containing plasmid exhibited decreased luciferase enzyme activity compared with the control.
Conclusion
MiR-1246 induced CSCC SiHa cell proliferation, invasion and migration. Preliminary evidence suggests that miR-1246 might promote CSCC tumorigenesis and progression by the suppression of its target gene THBS2.
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Acknowledgments
The present study was supported by grants from the Natural Science Foundation of Guangxi Zhuang Autonomous Region, China (2011GXNSFA018184, 2013GXNSFBA019130, 2013GXNSFBA019132. www.gxsti.net), and the Public Health self-financing project research project of Guangxi Zhuang Autonomous Region (Z2012071).
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The authors declare that they have no conflict of interest.
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Chen, J., Yao, D., Zhao, S. et al. MiR-1246 promotes SiHa cervical cancer cell proliferation, invasion, and migration through suppression of its target gene thrombospondin 2. Arch Gynecol Obstet 290, 725–732 (2014). https://doi.org/10.1007/s00404-014-3260-2
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DOI: https://doi.org/10.1007/s00404-014-3260-2