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[Study on interference therapy induced by epidermal growth factor receptor-antisense cDNA in signal transduction of
Jun-Ming Xian1, Guang-Yao Zhou, Chuan-Yu Liang
1Dept of Otolaryngology, West China Hospital, Sichuan University, Chengdu 610041, China.
Objective:
To investigate effect of interference therapy induced by epidermal growth factor receptor (EGFR)-antisense cDNA in signal transduction of Hep-2 laryngeal squamous cell carcinoma in vitro.
Methods:
AdEasy Vector System was used to construct the recombinant adenovirus vector sense/antisense-pAdEasy-EGFR. The recombinant adenovirus vector introduced EGFR-sense/antisense cDNA fragment into HEK293 cell. The purified recombinant adenovirus sense/antisense-pAdEasy-EGFR transfected Hep-2 cells in vitro. The inhibition of EGFR protein expression and proliferation of Hep-2 cells, the changes of cell cycle and DNA content in Hep-2 cells were examined by MTT, Western blot analysis, flow cytometry essay, and immunocytochemistry respectively.
Results:
The higher titre sense and antisense mRNA expression recombinant adenovirus containing 1,032 bp EGFR-cDNA was constructed and prepared successfully. When antisense-pAdEasy-EGFR was transferred into Hep-2 cells the inhibition of cell proliferation and EGFR protein expression in Hep-2 cells were investigated effectively.
Conclusion:
The antisense-pAdEasy-EGFR effectively interfere the Hep-2 signal transduction pathway and induce apoptosis which results in inhibiting proliferation of laryngeal squamous cell carcinoma.
Insights
Interference therapy using epidermal growth factor receptor (EGFR)-antisense cDNA effectively inhibits Hep-2 laryngeal squamous cell carcinoma proliferation. This approach targets the EGFR signal transduction pathway, offering a potential therapeutic strategy for this cancer.
Area of Science:
- Molecular Biology
- Oncology
- Gene Therapy
Background:
- Laryngeal squamous cell carcinoma (LSCC) is a significant global health concern.
- Epidermal growth factor receptor (EGFR) plays a crucial role in LSCC proliferation and survival.
- Targeting EGFR signaling presents a potential therapeutic avenue for LSCC.
Purpose of the Study:
- To investigate the efficacy of interference therapy using epidermal growth factor receptor (EGFR)-antisense cDNA in Hep-2 LSCC cells.
- To elucidate the impact of EGFR-antisense cDNA on LSCC signal transduction pathways in vitro.
Main Methods:
- Construction of recombinant adenovirus vectors carrying EGFR-sense/antisense cDNA using the AdEasy Vector System.
- Transfection of Hep-2 cells with recombinant adenovirus vectors.
- Assessment of EGFR protein expression, cell proliferation, cell cycle, and DNA content using MTT assays, Western blot, flow cytometry, and immunocytochemistry.
Main Results:
- Successful construction and preparation of high-titer recombinant adenovirus vectors encoding EGFR-cDNA.
- Demonstrated effective inhibition of Hep-2 cell proliferation and EGFR protein expression upon transfer of antisense-pAdEasy-EGFR.
- Observed alterations in cell cycle and DNA content indicative of growth inhibition.
Conclusions:
- Antisense-pAdEasy-EGFR effectively interferes with the Hep-2 cell signal transduction pathway.
- This interference induces apoptosis, leading to the inhibition of laryngeal squamous cell carcinoma proliferation.
- EGFR-antisense cDNA holds promise as a therapeutic agent for LSCC.
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