Identification Of The Expression Of Stat2 Gene And C3orf1 Gene In Esophageal Cancer And Lung Cancer

Human esophageal cancer is one of most common cancers worldwide and China has the highest mortality in the world. In our country esophageal cancer occurs at very high frequencies in certain areas, such as Lin-xian county in Henan province, northern parts of Jangsu province and Yangzhong in Zhenjiang. In order to explore the genes expressed differently in esophageal cancer, clinical specimens of esophageal cancer and lung cancer were collected from patients who received operations in Jangbin hospital, with each patient's tumor tissue and the corresponding normal tissue adjacent to tumor selected. In this study mRNA differential display (DD-PCR) was employed to search differently expressed fragments in esophageal cancer tissues, some of which were cloned and sequenced. The corresponding genes of those differently expressed fragments were validated by Real-time quantitative PCR. The following are what we have achieved:Ten cDNA fragments were excised by performing FDD-PCR reactions with esophageal cancer tissues and their respective normal tissues adjacent to the tumor, and two differential bands were obtained by carrying out the second PCR amplification. These two fragments were cloned and sequenced, one EST has highest homology (100%) with Homo sapiens signal transducer and activator of transcription 2 (stat2) gene, The ORF of stat2 gene was 2556 base pair long and encodes 851 amino acids. The molecular weight was about 97.9KD. The results of real-time quantitative PCR showed that the expression level of stat2 in esophageal cancer tissues (P<0.01, n=16) and lung cancer tissues (P<0.01, n=16) were both significantly higher than that in normal tissues.C3orf1 gene is a lung cancer related gene, which collected by mRNA differential display in our lab. This gene is an ORF, located in the third chromosome of human beings. The ORF of C3orf1 gene was 858 base pair long and encodes 285 amino acids. The molecular weight was about 31.4KD. The results of real-time quantitative PCR showed that the expression level of C3orf1 gene in lung cancer tissues (P<0.01, n=16) were higher than that in normal tissues. But the expression level of C3orf1 gene in esophageal cancer tissues (P<0.01, n=16) were mostly equal to that in normal tissues.C3orf1 gene was inserted into the baculovirus transfer plasmid pFastBac1, the recombinant plasmid pFastBac1+ C3orf1+gfp was obtained and transformed into the competent cells of DH10Bac, then recombinant virus DNAs were transfected into Bm cells. Western-Blot was used to test the expression products of the recombinant viruses. The results showed there was a special band at about 57kD, indicated that C3orf1+gfp successfully expressed in Bm cell.