[Isolation and identification of cDNA sequence differentially expressed in human lung carcinoma].
Hailong Xie, You-jun Li, Zhuchu Chen, Yongjun Guan, Chun-Mei He
Abstract
Hailong Xie, You-jun Li, Zhuchu Chen, Yongjun Guan, Chun-Mei He
Abstract
BACKGROUND & OBJECTIVE: The molecular mechanism of the pathogenesis of lung carcinoma is unclear; because key genes related to lung carcinoma have not been identified. This study was designed to isolate and identify differentially expressed cDNA sequences in human lung carcinoma for cloning lung carcinoma-related genes. METHOD: Using mRNA differential display method combined with cloning and reserve Northern dot blot and sequencing and RT-PCR, differentially expressed cDNA sequences were isolated between two lung cancer cell lines, two adult lung carcinoma tissue and paired normal tissue of trachea epithelia. RESULT: Sixteen differentially expressed cDNA fragments were isolated, subsequent cloning of sixteen differentially expressed cDNA fragments were confirmed by reverse Northern dot blot and sequencing and BLAST analysis. Two of these were shown to be novel gene sequences that had not been reported. Eight of the remaining cDNA sequences homologize to known genes; additionally two differentially expressed cDNA sequences were confirmed by RT-PCR. CONCLUSION: Using mRNA differentially display, The authors had successfully isolated ten differentially expressed cDNA sequences from human lung carcinoma, these cDNA sequences might be involved in the pathogenesis of lung carcinoma.
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BACKGROUND & OBJECTIVE: The molecular mechanism of the pathogenesis of lung carcinoma is unclear; because key genes related to lung carcinoma have not been identified. This study was designed to isolate and identify differentially expressed cDNA sequences in human lung carcinoma for cloning lung carcinoma-related genes. METHOD: Using mRNA differential display method combined with cloning and reserve Northern dot blot and sequencing and RT-PCR, differentially expressed cDNA sequences were isolated between two lung cancer cell lines, two adult lung carcinoma tissue and paired normal tissue of trachea epithelia. RESULT: Sixteen differentially expressed cDNA fragments were isolated, subsequent cloning of sixteen differentially expressed cDNA fragments were confirmed by reverse Northern dot blot and sequencing and BLAST analysis. Two of these were shown to be novel gene sequences that had not been reported. Eight of the remaining cDNA sequences homologize to known genes; additionally two differentially expressed cDNA sequences were confirmed by RT-PCR. CONCLUSION: Using mRNA differentially display, The authors had successfully isolated ten differentially expressed cDNA sequences from human lung carcinoma, these cDNA sequences might be involved in the pathogenesis of lung carcinoma.
Key concepts: Complementary DNA, Biology, Molecular biology, Cloning (programming), Northern blot, cDNA library, Gene, Molecular cloning