[The effect of shRNA targeting hTERT on telomerase and the expression of PCNA and Caspase-3 in nasopharyngeal carcinoma cells].
OBJECTIVE
Analysis of the correlation between telomerase and expression of its related proteins may provide insight into the molecular mechanism of nasopharyngeal carcinogenesis. Here, we investigate the effect of short hairpin RNA (shRNA) specific for human telomerase reverse transcriptase (hTERT) mRNA on the expression of proliferating cell nuclear antigen (PCNA) and Caspase-3 in nasopharyngeal carcinoma (CNE) cells.


METHODS
shRNA expression vectors targeting the mRNA of hTERT were constructed. Cells were treated with the constructed expression vectors and telomerase activity was measured by telomeric repeat amplification ELISA (TRAP-ELISA). Cell viability was examined using the MTT assay. Cell apoptosis was detected by TUNEL method. The expression of PCNA and Caspase-3 proteins, was determined by Western blotting.


RESULTS
shRNA specific for hTERT mRNA significantly inhibited telomerase activity, suppressed cell viability and induced apoptosis in CNE cells. In addition, the expression of PCNA was inhibited, while the expression of Caspase-3 was up-regulated.


CONCLUSIONS
Our results suggest that shRNA directed against hTERT inhibits cell viability by regulating telomerase activity and its related protein expression in NPC cells. Therefore, RNA-interfering technology may be a promising strategy for the treatment of nasopharyngeal cancer.
Shen
Yongzhong
Y
Department of Otolaryngology, Remin Hospital of Qichun County, Qichun, 435000, China.
Wang
Yan
Y
Chen
Shiming
S
Xiao
Bokui
B
Su
Jun
J
Tao
Zezhang
Z
D053148:Caspase 3 / Q000378:metabolism*; D045744:Cell Line, Tumor; D002470:Cell Survival; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000235:genetics* / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D034741:RNA, Small Interfering; D019098:Telomerase / Q000235:genetics*; D014162:Transfection
D018809:Proliferating Cell Nuclear Antigen; D034741:RNA, Small Interfering; C509186:TERT protein, human; D019098:Telomerase; C505407:CASP3 protein, human; D053148:Caspase 3