Serial Analysis of Gene Expression in Immortalized BEP2D Cells and Malignant Transformed BEP2D Cells Induced by α-particle
10.3321/j.issn:1000-467X.2001.01.003
- VernacularTitle:SAGE方法分析永生化BEP2D细胞及α粒子诱发恶性转化BEP2D细胞的基因表达
- Author:
Shi-Li GE
1
;
Gang LI
;
Wei CHEN
;
Tie-Zhu LOU
;
De-Chang WU
Author Information
1. Beijing Institute of Radiation Medicine
- From:Chinese Journal of Cancer
2001;20(1):12-17
- CountryChina
- Language:Chinese
-
Abstract:
Objective： The authors used the serial analysis of gene expression (SAGE) method to analyze transcripts present in the immortalized BEP2D cells and the malignant transformed BEP2D cells. Methods： As a step toward understanding the complex gene expression differences between the immortalized BEP2D cells and the malignant transformed BEP2D cells induced by α particle,SAGE method was introduced into this experiment. Technological methods included total RNA extraction, mRNA isolation, full length of dscDNA synthesis, PCR, transformation and sequencing. SAGE SoftWare was used to analyze tag sequences and compare the abundance of tags between two SAGE libraries. Results： Two independent SAGE libraries were constructed from the immortalized BEP2D cells and the malignant transformed BEP2D cells induced by 1.5 Gy α particles. A total of 2 331 SAGE tags were identified for the sequences, representing 252 unique transcripts. Though up to now the obtained information is limited, comparison of the two SAGE libraries indicated a remarkable similarity in the expression profiles. Of the 252 transcripts detected, 12 transcripts (4.8％ ) matched no reliable known genes in UniGene library. Combination of Northern blot hybridization, the expression level of TGF β induced Smad7 gene in the malignant transformed cells was higher than that in the immortalized cells. Conversely, Chemokine receptor CCR11 gene was expressed at lower levels in the malignant transformation cells. Conclusion： (1) Out of results given by SAGE,the tendency of the abundance of gene expression and the gene expression differentiation between two cell lines were described. (2) The expression level of TGF β induced Smad7 gene in the malignant transformed BEP2D cells was higher than that in the immortalized cells and chemokine receptor CCR11 gene was expressed at lower levels in the malignant transformed BEP2D cells. (3)SAGE was a powerful method in a quantitative and simultaneous analysis of a large number of transcripts in any particular cell system, especially in defining functions of known genes.
