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[Molecular cloning of genes differentially expressed in human lung squamous cell carcinoma using suppression
Chenyang Shen1, Jun Liu, Danlei Wang
1Department of Thoracic Surgery, The People's Hospital, Peking University, Beijing 100044, P.R.China.
Background:
To clone and identify genes differentially expressed in human lung squamous cell carcinoma (LSCC).
Methods:
A subtracted cDNA library of human LSCC was constructed by suppression subtracted hybridization (SSH) method. After screening, the subtracted library clones representing mRNAs that were truly differentially expressed in LSCC but not in its adjacent non cancerous tissues were selected to identify by RT-PCR and DNA sequencing were performed. Nucleic acid homology searches were performed using the BLAST program.
Results:
By this technique, 10 differentially expressed gene cDNA fragments of LSSC were obtained. Two were novel and eight were already known genes.
Conclusions:
SSH is a useful technique with high sensitivity for the detection of differential genes expression in LSCC and an effective method to clone novel genes.
Insights
This study identified 10 differentially expressed genes in human lung squamous cell carcinoma (LSCC) using suppression subtracted hybridization (SSH). SSH is a sensitive method for discovering novel genes in LSCC.
Area of Science:
- Molecular Biology
- Genomics
- Cancer Research
Context:
- Human lung squamous cell carcinoma (LSCC) is a major subtype of lung cancer.
- Identifying genes that are differentially expressed is crucial for understanding LSCC pathogenesis.
- Existing methods may have limitations in sensitivity for novel gene discovery.
Purpose:
- To clone and identify genes with altered expression levels in LSCC compared to adjacent non-cancerous tissues.
- To utilize suppression subtracted hybridization (SSH) for efficient gene discovery.
- To validate differentially expressed genes using RT-PCR and DNA sequencing.
Summary:
- Suppression subtracted hybridization (SSH) was employed to construct a subtracted cDNA library from LSCC tissues.
- Screening identified 10 differentially expressed gene cDNA fragments, including 2 novel genes and 8 known genes.
- RT-PCR and DNA sequencing confirmed the differential expression and identity of selected clones.
Impact:
- SSH proves to be a highly sensitive technique for detecting differential gene expression in LSCC.
- This method facilitates the effective cloning of novel genes relevant to LSCC development.
- The identified genes may serve as potential biomarkers or therapeutic targets for LSCC.
