Inhibitory effect of interference hTERT and TRF2 gene on the growth of breast cancer MCF-7 cells / 中华肿瘤杂志
Chinese Journal of Oncology
;
(12): 93-97, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-295173
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of combined gene therapy with interference hTERT and TRF2 gene on the treatment of breast cancer.</p><p><b>METHODS</b>Recombinant adenovirus rAd-hTERT and rAd-TRF2 expressing siRNA-hTERT and siRNA-TRF2 was constructed, and the vectors were transfected into MCF-7 cells. Than the expressions of hTERT and TRF2 proteins were detected by Western blot, the inhibition of MCF-7 cell proliferation by MTT colorimetry, and the changes of MCF-7 cell cycle by flow cytometry and the colony forming ability of MCF-7 cells by clone form test.</p><p><b>RESULTS</b>At 48 h after transfection, the relative expression amounts of hTERT protein of the PBS control group, rAd-blank group, rAd-HK control group, rAd-hTERT group, rAd-TRF2 group and rAd-hTERT and rAd-TRF2 group were 1.00, 0.94 +/- 0.02, 0.95 +/- 0.04, 0.18 +/- 0.04, 0.95 +/- 0.01 and 0.18 +/- 0.04, respectively. The relative expression amounts of TRF2 protein were 1.00, 1.01 +/- 0.08, 0.96 +/- 0.02, 0.95 +/- 0.08, 0.22 +/- 0.01 and 0.26 +/- 0.02, respectively. After transfection of rAd-hTERT or rAd-TRF2 into MCF-7 cells separately, the inhibition rate of cell proliferation was only 54.6% and 48.4%, there was 8.9% +/- 1.2% or 9.2% +/- 2.3% of MCF-7 cells into M phase, 66.4% +/- 1.5% or 64.6% +/- 1.9% of MCF-7 cells was arrested at G(0)/G(1) phase, and the cell colony forming ability was decreased significantly (cell colony number from 100 in PBS control group down to 41.3 +/- 5.1 and 43.7 +/- 6.4). But after transfection by rAd-hTERT and rAd-TRF2 simultaneously, the inhibition rate of cell proliferation was about 82.1%, and M phase cells was significantly reduced to 4.4% +/- 1.2%. Large numbers of cells were arrested at G(0)/G(1) phase (81.4% +/- 1.3%), and the cell colony forming ability was more significantly decreased (cell colony number there were only 29.2 +/- 3.9).</p><p><b>CONCLUSION</b>More effective effect of tumor gene therapy can be achieved by combination of interference hTERT and TRF2 genes as compared with interference by either of the single gene alone.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Breast Neoplasms
/
Transfection
/
Tumor Stem Cell Assay
/
Genetic Therapy
/
Gene Expression Regulation, Neoplastic
/
Cell Cycle
/
Adenoviridae
/
Telomerase
/
Telomeric Repeat Binding Protein 2
Limits:
Female
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Oncology
Year:
2010
Type:
Article
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