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Updated: Jan 23, 2026
Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein
Published on: June 12, 2018
[Influence of RNA interference targeting against human telomerase reverse transcriptase on expression of C-myc
Huaming Chi1, Zezhang Tao, Shiming Chen
1Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, China.
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.
Method:
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.
Result:
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).
Conclusion:
The inhibition of hTERT expression could increase the expression of C-myc protein in Hep-2 cells.
Insights
Inhibiting human telomerase reverse transcriptase (hTERT) using RNA interference in Hep-2 larynx cancer cells led to increased C-myc protein expression. This finding suggests a potential link between hTERT and C-myc in laryngeal cancer progression.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Context:
- Human telomerase reverse transcriptase (hTERT) plays a crucial role in cell proliferation and immortality.
- C-myc protein is an oncogene frequently dysregulated in various cancers, including laryngeal cancer.
- Understanding the relationship between hTERT and C-myc is vital for developing targeted cancer therapies.
Purpose:
- To investigate the effect of inhibiting hTERT on C-myc protein expression in the Hep-2 larynx cancer cell line.
- To utilize RNA interference (RNAi) technology for targeted gene silencing of hTERT.
- To analyze changes in hTERT and C-myc protein levels following hTERT inhibition.
Summary:
- hTERT gene silencing was achieved using a synthesized shRNA plasmid (pshRNA1) in Hep-2 cells.
- RT-PCR and Western Blot analysis confirmed significant downregulation of hTERT mRNA and protein expression.
- A significant upregulation of C-myc protein expression was observed post-hTERT inhibition (P < 0.01).
Impact:
- The study demonstrates that inhibiting hTERT expression can lead to increased C-myc protein levels in Hep-2 cells.
- This finding may reveal a novel regulatory pathway involving hTERT and C-myc in laryngeal tumorigenesis.
- Further research could explore targeting the hTERT-C-myc axis for therapeutic interventions in larynx cancer.
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