The effect of TET1 on the proliferation of renal cancer 786-O cells and its related mechanism
10.3781/j.issn.1000-7431.2012.12.003
- Author:
Su-Hong XIE
1
Author Information
1. Department of Clinical Laboratory
- Publication Type:Journal Article
- Keywords:
Cell proliferation;
Kidney neoplasms;
RNA interference;
Suppressor of zeste 12;
Ten-eleven translocation
- From:
Tumor
2012;32(12):962-968
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To silence the expression of TET 1 (ten-eleven translocation 1) gene by siRNA (small interfering RNA) and to investigate its impact on the proliferation of human renal cancer 786-O cells and explore its possible mechanism. Methods: The TET1-siRNA targeting human TET 1 gene was transfected into 786-O cells by LipofectAMINE 2000.Three groups were designed as blank control, negative control and siRNA interference (TET1-siRNA) in this study. The expression levels of TET1, SUZ12 (suppressor of zeste 12), EZH2 (enhancer of zeste homolog 2) and EED (embryonic ectoderm development) mRNAs and proteins after transfection with TET1-siRNA were determined by real-time fluorescence quantitative-PCR and Western blotting, respectively. The MTT method, colony formation assay and flow cytometry were performed to detect the proliferation ability, cell colony formation rate and cell cycle distribution of 786-O cells, respectively. Results: As compared with the blank control, the expression levels of TET1 mRNA and protein in 786-O cells were reduced by (85.13±0.03)% and (56.41±0.09)% respectively after transfection with TET1-siRNA (P < 0.05). The proliferation of 786-O cells was inhibited and the ability of colony-formation was also weakened. The cell cycle of 786-O cells was arrested at G0/G1 phase (P < 0.05). The expression levels of SUZ12 mRNA and protein were reduced significantly (P < 0.05), meanwhile the expression levels of EED and SUZ12 proteins had no change (P > 0.05). Conclusion: TET1-siRNA can down-regulate the expression levels of TET1 mRNA and protein in human renal cancer 786-O cells, suppress the proliferation of 786-O cells, and block the cells at G1 phase. This mechanism may be associated with the recruitment of PRC2 (polycomb repressive complex 2) influenced by TET1 which regulates the expression of SUZ12. Copyright © 2012 by TUMOR.
