Suppression Subtractive Hybridization Identified Genes Differentially Expressed in a Multidrug Resistance Cell Line of Human Lung Adenocarcinoma
Jie Chen
Abstract
Jie Chen
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.
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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.
Key concepts: Complementary DNA, Suppression subtractive hybridization, Biology, cDNA library, Molecular biology, Gene, Multiple drug resistance, Genetics