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[Cloning and primary expression analyses of down-regulated cDNA fragment in human gastric cancer]
1Department of Gastroenterology, General Hospital of Chinese PLA, Beijing, 100853 P.R. China. wanggangshi@hotmail.com
Objective:
To identify human gastric cancer related genes.
Methods:
Specimens of paired tumor, paratumor and normal gastric mucosa tissues were collected from five patients (male 3, female 2, with average age 48.8 +/- 18.1 years) who suffered from stomach antrum adenocarcinoma. Total RNA samples were extracted from these specimens, then studied by fluorescent differential display reverse transcription polymerase chain reaction (DDRT- PCR) analysis. The differentially expressed bands of interest were recovered, purified and cloned, then they were analyzed by sequencing, Northern blot and RT-PCR. Through BLAST, the sequencing results were compared with GenBank database for homology analysis.
Results:
One of the interesting cDNA bands expressed much lower in all five tested tumor samples than in their normal and paratumor counterparts. This band was named W4. Northern blot analysis showed a consistent result with that of DDRT-PCR. W4 was cloned into pGEM-T easy vector. Sequence analysis showed that W4 consists of 712bp, this sequence was named W44. BLAST analysis revealed that W44 has extremely low sequence identity with any genes from GenBank and any sequences from EST database. This sequence data was submitted to GenBank with accession No. AF150631. RT-PCR analysis showed that W4 was expressed much lower in 11/15 gastric cancer tissue than in paratumor and normal samples.
Conclusion:
A novel cDNA sequence related to human stomach adenocarcinoma was identified.
Insights
Researchers identified a novel gene, W44, significantly downregulated in human gastric cancer tissues. This discovery offers new insights into stomach adenocarcinoma development and potential diagnostic markers.
Area of Science:
- Molecular biology
- Genetics
- Oncology
Context:
- Gastric cancer is a significant global health concern.
- Identifying novel genes involved in tumorigenesis is crucial for understanding disease mechanisms.
Purpose:
- To identify novel genes associated with human gastric cancer.
- To characterize the expression profile of differentially expressed genes in gastric adenocarcinoma.
Summary:
- Fluorescent differential display reverse transcription polymerase chain reaction (DDRT-PCR) was used to analyze gene expression in paired tumor, paratumor, and normal gastric mucosa tissues.
- A novel cDNA sequence, W44 (GenBank accession No. AF150631), was identified and found to be significantly downregulated in most gastric cancer samples.
- Northern blot and RT-PCR confirmed the reduced expression of W44 in gastric cancer tissues.
Impact:
- Discovery of a novel gene potentially involved in gastric cancer development.
- Provides a new candidate for diagnostic or therapeutic strategies in stomach adenocarcinoma.
- Contributes to the understanding of molecular alterations in gastric tumorigenesis.