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Objectives The chief objective of this study was to identify the miRNAs targeting Fos , a well-recognized proto-oncogene that is commonly overexpressed in cervical cancer, and its biological significance on the cellular behaviors of HeLa, a cervical cancer cell. Materials and Methods We initially an...
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Published in | International journal of gynecological cancer Vol. 24; no. 7; pp. 1165 - 1172 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
Oxford
BMJ Publishing Group LTD
01.09.2014
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Subjects | |
Online Access | Get full text |
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Summary: | Objectives
The chief objective of this study was to identify the miRNAs targeting
Fos
, a well-recognized proto-oncogene that is commonly overexpressed in cervical cancer, and its biological significance on the cellular behaviors of HeLa, a cervical cancer cell.
Materials and Methods
We initially analyzed the 3′untranslated region (3′UTR) of
Fos
and screened the potential miRNAs targeting
Fos
using 3 bioinformatical Web sites. Luciferase reporter assay, real-time polymerase chain reaction, and Western blotting were used to validate the binding of chosen miRNA (miR-101) on the 3′UTR of
Fos
and the downstream regulation on its mRNA and protein levels. Furthermore, flow cytometry along with the
Fos
rescue strategy was applied to analyze the modulation of cell cycle of HeLa cells by miR-101.
Results
Among these predicted candidate miRNAs, miR-101 was the miRNAs preferred by all the 3 used Web sites. The results of luciferase reporter assay, real-time polymerase chain reaction, and Western blotting demonstrated that miR-101 directly targeted on the 3′UTR of
Fos
and down-regulated the expression of
Fos
at mRNA and protein levels. Furthermore, cell cycle analysis showed that miR-101 arrests G1-to-S phase transition of HeLa cells, at least partially by targeting
Fos
.
Conclusions
We concluded that by targeting the proto-oncogene
Fos
, miR-101 is involved in G1-to-S phase transition in cervical cancer cells in vitro and might provide a new approach for the pharmacological interference node in cervical cancer treatment. |
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ISSN: | 1048-891X 1525-1438 |
DOI: | 10.1097/IGC.0000000000000187 |