Spaceflight alters the gene expression profile of cervical cancer cells / 癌症

Our previous study revealed that spaceflight induced biological changes in human cervical carcinoma Caski cells. Here, we report that 48A9 cells, which were subcloned from Caski cells, experienced significant growth suppression and exhibited low tumorigenic ability after spaceflight. To further understand the potential mechanism at the transcriptional level, we compared gene expression between 48A9 cells and ground control Caski cells with suppression subtractive hybridization (SSH) and reverse Northern blotting methods, and analyzed the relative gene network and molecular functions with the Ingenuity Pathways Analysis (IPA) program. We found 5 genes, SUB1, SGEF, MALAT-1, MYL6, and MT-CO2, to be up-regulated and identified 3 new cDNAs, termed B4, B5, and C4, in 48A9 cells. In addition, we also identified the two most significant gene networks to indicate the function of these genes using the IPA program. To our knowledge, our results show for the first time that spaceflight can reduce the growth of tumor cells, and we also provide a new model for oncogenesis study.

Construction of eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment gene G22 and its expression / 中南大学学报(医学版)

OBJECTIVE@#To construct a eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment (ScFv) gene G22, and to identify its expression in rectal cancer cells (CMT-93).@*METHODS@#The G22 gene was ligated into the sites of EcoRI and NotI of eukaryotic expression vector pcDNA3.1(+). After the identification and DNA sequencing, the recombinant plasmid pc DNA3.1(+)-G22 was stably transfected into CMT-93 cells, and the expression of G22 was detected by Western blot, flow cytometry and immunofluorescence staining.@*RESULTS@#Restriction enzyme digestion and DNA sequencing showed that the recombinant plasmid contained the human nasopharyngeal carcinoma anti-idiotype antibody ScFv gene G22. Transfection experiment verified that G22 gene could be expressed in CMT-93 cells in the right way.@*CONCLUSION@#The eukaryotic expression vector containing the human nasopharyngeal carcinoma anti-idiotype antibody ScFv gene G22 is successfully constructed and expressed, which is the basis for further study of its DNA vaccine.

Detection of autoantibodies in the sera of nasopharyngeal carcinoma patients / 中南大学学报(医学版)

OBJECTIVE@#To investigate whether there are autoantibodies to nasopharyngeal carcinoma (NPC) in the sera of patients and to find new NPC biomarkers.@*METHODS@#Cell plate of Epstein Barr virus negative NPC cell line CNE1 was prepared, and difference between 32 NPC patient sera and 54 normal sera was analyzed by ELISA. We extracted the total protein of CNE1, and analyzed whether there were specific proteins to react with NPC sera by Western blot.@*RESULTS@#The average absorbance values of serum antibody in NPC patients (0.904+/-0.032) were significantly higher than those of normal serum antibody absorbance values (0.736+/-0.028) (P< 0.01). The analysis with Western blot showed there were positive bands,and some of these were unanimous bands, but the intensity increased, and some of these were new bands compared with the normal sera. These positive bands may be NPC tumor-associated antigens or NPC tumor-specific antigens.@*CONCLUSION@#Autoantibodies that react with NPC exist in the sera of NPC patients, but they do not react with Epstein-Barr virus. It provides the basis to seek the tumor biomarkers in NPC sera.

Biological properties of Caski cell lines induced by exposing to the space environment / 中南大学学报(医学版)

OBJECTIVE@#To investigate the biological properties of Caski cell lines induced by exposing to the space environment.@*METHODS@#Caski cells were carried in "Shen Zhou IV" airship. After 7 days of spaceflight, cells survived and were monocoloned, and the experimental methods such as cell morphological observation, the cell proliferation assay, flow cytometry cell cycle analysis, the soft agar assay, and tumorigenesis assay were used to analyze cell growth characteristics and malignant phenotypes.@*RESULTS@#Altogether 1440 strains subclonal cell lines were established and 4 strains were screened. Compared with the control group, mutated cells appeared to have multiple cell morphological changes. Strains numbered 44F10 and 17E3 were screened due to their increased cell proliferation and tumorigenesis, and their cell cycles were induced to progress from G(1) to S phase, while strains 48A9 and 31F2 were opposite to 44F10 and 17E3 in cytological events. The average population double time of ground nomal control group, ground simulant control group, strains numbered 44F10, 17E3, 48A9 and 31F2 groups were 56.54, 58.44, 52.96, 51.46, 101.76 and 88.47h, respectively; compared with the control group, the average double time of strains numbered 44F10 and 17E3 was decline, but with no statistical significance. However, compared with the control groups, the average double time of 48A9 and 31F2 was significant increased (P<0.05). The colony formation rates were 9.7%, 9.3%, 14.7%, 12.1%, 0 and 0.1%, respectively, and the difference between the ground control groups and the other groups was significant (P<0.01); 6 groups of above-mentioned Caski cells were inoculated subcutaneously in Babl/c nude mice respectively. Forty-seven days later, the formed tumors in the nude mice were statistically analyzed and tested. The average weight of tumors of the above-mentioned groups was 0.066, 0.066, 0.175, 0.249, 0.011 and 0.018g. The difference between the ground control groups and other groups was significant (P<0.05).@*CONCLUSION@#Spaceflight may affect the physiological characteristics of tumor cells and the variation is complicated.

Construction of cDNA library from NPC tissue and screening of antigenic genes / 生物工程学报

To obtain the NPC-associated antigens, a powerful new method, SEREX (serological identification of antigen by recombinant cDNA expression library), was used for identifying the antigens eliciting humoral immune response. Before performing serological analysis, a high quality cDNA library derived from human nasopharyngeal carcinoma (NPC) tissue was constructed. The primary library consisted of 3.64 x 10(6) recombinants and the recombinant rate was 94%. For better preserving the cDNA library, it was amplified. As a result, the titer of the amplified cDNA library was 3.8 x 10(9) pfu/mL. With SEREX method, immunoscreening for the detection of reactive clones in the human NPC tissue cDNA library was performed with autologous serum. As a result, 23 positive clones encoding antigenic genes were obtained after immunoscreening, and the nucleotide sequences of cDNA inserts were determined and analyzed with BLAST software in GenBank. Results showed that the 23 reactive clones were derived from 16 different genes. 10 of 16 genes had high homologous to the genes known in GenBank, such as RPL31, S100 A2, MT2A, etc. However, there were also 6 genes with low homology to the genes known in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be revealed by further research.