Cyclin-dependent kinase 2-associated protein 1(CDK2AP1), a cell growth inhibitory factor, is abnormally expressed in cancer cells, and might be implicated in the development of lung cancer. However, no studies on the function of CDK2AP1 in human lung cancer have been yet reported. In this study, overexpressing lentiviral vectors containing full-length CDK2AP1 cDNA and CDK2AP1 shRNA(short hairpin RNA) were constructed. Our results show that infecting A549 cells with lentivirus containing CDK2AP1 shRNA or full-length CDK2AP1 cDNA results in significantly down- or up-regulated the expression of CDK2AP1, respectively, thereby providing reliable tools for studying the function of CDK2AP1 in pulmonary carcinogenesis. Our data of MTT assay and flowcytometric analysis demonstrate that CDK2AP1 plays an important role in the proliferation/growth and cell cycling of A549 cells in vitro, and further investigation into its underlying mechanism of pulmonary carcinogenesis is needed.
GAO NanZHANG Xing-yiJIANG RuiWANG GuanLIJin-dongJINCheng-yanSUN Mei
Differential proteins expressing in ectopic and eutopic endometria were investigated by means of proteomic analysis. Five patients in secretary phase were diagnosed as endometriosis by laparoscopy. The five ectopic endometria(two at stage Ⅱ, two at stage Ⅲ and one at stage IV) and five eutopic endometria were surgically excised. One-dimensional electrophoresis coupled with liquid chromatography and mass spectrometry was used to screen and identify differential proteins. Three differential bands in one-dimensional electrophoresis were resolved by liquid chromatography and mass spectrometry and 14 up-regulated proteins were identified, including collagen α-1, α-2, α-3(VI), α-1(XIV) chain, actin, annexin A2, EMILIN-1, ferritin light polypeptide variant, fucosyltransferase 10, myosin-9, protein S100-A9, KIAA1783 protein, and two hypothetical proteins. Our data provides a list of potential biomarkers for endometriosis. The identifications may be used to develop new diagnoses for endometriosis.
ZHANG Li-huiLIU Hong-yanLI He-lianZHANG Xing-yiSUN Mei
To construct overexpression lentiviral-based vector carrying rat B7x gene,B7x gene precursor sequences amplified by polymerse chain reaction(PCR) were ligated with pLVTHM to generate pLVTHM-B7x gene expression lentiviral-based vector.The positive clones were selected to be submitted to DNA sequencing.HEK293T cells were co-transfected with pLVTHM-B7x and two packaging plasmids psPAX2 and pMD2G to produce lentivirus which express B7x gene.The mRNA expression levels of B7x gene and virus titer were detected by real-time PCR in HEK293T cells.The B7x protein levels were detected by Western blot analysis in HEK293T cells.The identification of restriction enzyme digestion and DNA sequencing confirmed that pLVTHM-B7x lentiviral-based vector was successfully constructed.Green fluorescence was observed in HEK293T packaging cells by means of an inverted fluorescence microscope and the virus titer measured was 2×108 TU/mL.It will establish the foundation for studing deeply the biological function of B7x to construct successfully B7x expression lentiviral-based vector.
SUN MeiJIANG RuiSUN Li-huaLI Jin-dongWANG BinJIN Cheng-yanZHANG Xing-yi
