[Influence of RNA interference targeting against human telomerase reverse transcriptase on expression of C-myc protein].
OBJECTIVE
To investigate the effect of inhibiting human telomerase reverse transcriptase (hTERT) on expression of C-myc protein by RNA interference (RNAi) in the larynx cancer cell line, Hep-2.


METHODS
The primary structures of hTERT cDNA were found in GeneBank. Then the structure analyses were done according to the strategy of RNAi, which determined the specific base sequences to design shRNA plasmid. One type of plasmid, pshRNA1, involved in fluorescein gene was synthesized based on the specific base sequence. Control pshRNA2-a random sequence-were also constructed. METAFECTENE was used as the transfect ion reagent. Cells were treated daily with pshRNA1-2 or normal culture medium respectively. After administration of pshRNA1-2, hTERT mRNA was detected by RT-PCR, hTERT protein and C-myc protein were examined by Western Blot.


RESULTS
The expression of hTERT mRNA and protein were both significantly decreased after treated by pshRNA1 (P < 0.05). The expression of C-myc protein was significantly increased after treated by pshRNA1 (P < 0.01).


CONCLUSIONS
The inhibition of hTERT expression could increase the expression of C-myc protein in Hep-2 cells.
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, China.
Huaming
H
Chi
Zezhang
Z
Tao
Shiming
S
Chen
Bokui
B
Xiao
Hanzhang
H
Zhan
D045744:Cell Line, Tumor; D005822:Genetic Vectors; D006801:Humans; D007822:Laryngeal Neoplasms / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019098:Telomerase / Q000235:genetics*; D014162:Transfection
D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019098:Telomerase