To screen the over differentially expressed genes in carcinoma induced by BPDE-transformed 16HBE cells （16HBE-C cells）. Methods The suppression subtractive hybridization （SSH） method was performed to profile diffe...To screen the over differentially expressed genes in carcinoma induced by BPDE-transformed 16HBE cells （16HBE-C cells）. Methods The suppression subtractive hybridization （SSH） method was performed to profile differentially expressed genes between 16HBE-C cells and 16HBE cells. The cDNA fragments of differentially expressed genes were inserted into TA cloning vector and transformed competent E. coli strain. Positive clones were randomly picked up and identified by the colony PCR method. Dot blot was used to test the same source with the tester. The differentially expressed cDNA fragments were sequenced and compared with known genes and EST database in Genbank. Results Eight known genes were over-expressed in 16HBE-C cells including eukaryotic translation elongation factor 1 alpha 1, HIF-1 responsive RTP801, ribosomal protein L10 （RPL10）, ribosomal protein S29 （RPS29）, mitochondrion related genes, and laminin receptor 1. Three differentially expressed cDNA fragments could not be matched to the known genes but to the EST database. Conclusion The SSH method can detect differentially expressed genes between 16HBE-C and 16HBE cells. BPDE-induced carcinogenesis may be related to alteration of at least eight known genes and three unknown genes. These expression data provide a clue to further cloning novel genes and studying functions in BPDE-induced carcinoma.展开更多
基金This work was supported by the grants from the National Natural Science Foundation of China (No. 30271111), the Provincial Natural Science Foundation of Guangdong (No. 020734) and National Key Basic Research and Development Program (2002CB512905).
文摘To screen the over differentially expressed genes in carcinoma induced by BPDE-transformed 16HBE cells （16HBE-C cells）. Methods The suppression subtractive hybridization （SSH） method was performed to profile differentially expressed genes between 16HBE-C cells and 16HBE cells. The cDNA fragments of differentially expressed genes were inserted into TA cloning vector and transformed competent E. coli strain. Positive clones were randomly picked up and identified by the colony PCR method. Dot blot was used to test the same source with the tester. The differentially expressed cDNA fragments were sequenced and compared with known genes and EST database in Genbank. Results Eight known genes were over-expressed in 16HBE-C cells including eukaryotic translation elongation factor 1 alpha 1, HIF-1 responsive RTP801, ribosomal protein L10 （RPL10）, ribosomal protein S29 （RPS29）, mitochondrion related genes, and laminin receptor 1. Three differentially expressed cDNA fragments could not be matched to the known genes but to the EST database. Conclusion The SSH method can detect differentially expressed genes between 16HBE-C and 16HBE cells. BPDE-induced carcinogenesis may be related to alteration of at least eight known genes and three unknown genes. These expression data provide a clue to further cloning novel genes and studying functions in BPDE-induced carcinoma.
