Suppression Subtractive Hybridization Identified Genes Differentially Expressed in a Multidrug Resistance Cell Line of Human Lung Adenocarcinoma
10.3321/j.issn:1000-467X.2001.04.002
- VernacularTitle:肺腺癌多药耐药细胞特异表达基因的克隆与鉴定
- Author:
Jie CHEN
1
;
Gui-Sheng QIAN
;
Gui-Jun HUANG
;
Wei XIONG
;
Jing LI
Author Information
1. 第三军医大学附属新桥医院
- From:Chinese Journal of Cancer
2001;20(4):348-353
- CountryChina
- Language:Chinese
-
Abstract:
Objective: The aim of this study was to clone and screen multidrug resistance related gene of human adenocarcinoma cell. Methods: The suppression subtractive hybridization (SSH) was performed on human adenocarcinoma multidrug resistance cell line (SPC-A-1/CDDP, as tester) and human adenocarcinoma cell line (SPC-A-1, as driver). After the subtracted cDNA library being constructed, the dot blots was used to screen the subtracted cDNA library with forward and reverse-subtracted cDNA probes. The differentially expressed cDNA fragments in SPC-A-1/CDDP was sequenced and analyzed through Genbank with Blast search. The novel cDNA sequences were analyzed by Northern blots. Results: A high quality subtracted cDNA library was constructed. Twenty-three differentially expressed cDNA fragments in SPC-A-1/CDDP were identified. Two of them were novel cDNA sequences and the others had 93％－100％ homology with the known genes respectively. Northern blots indicated the novel cDNA sequences only expressed in SPC-A-1/CDDP cell. Conclusion: The novel cDNA sequences might be multidrug resistance related genes in human lung adenocarcinoma. SSH is a powerful technique to identify differentially expressed genes.
