Résumé
<p><b>OBJECTIVE</b>Construct the gene library of apoptosis related genes in acute promyelocytic leukemia (APL) cell line NB4 cells treated by arsenic trioxide to clarify the apoptotic mechanism of NB4 cells.</p><p><b>METHOD</b>APL cell line NB4 cells treated with or without arsenic trioxide for 24 hours. Total RNA was extracted and suppress subtractive hybridization (SSH) was conducted according to the manual. With the cDNA of the apoptosis cells as the tester and that of control cells as the driver, forward and reverse hybridization was performed. Differentially expressed genes were linked with pGEM-Teasy cloning vector and transformed into E. coli DH5alpha. The positive clones were screened by blue and white spot. PCR were used to amplify these genes.</p><p><b>RESULT</b>The subtractive cDNA libraries related with apoptosis of NB4 cells were successfully constructed.</p><p><b>CONCLUSION</b>The constructed subtractive libraries are suitable for further study on the functional genes associated with apoptosis ofNB4 cells induced by arsenic trioxide.</p>
Sujets)
Humains , Apoptose , Génétique , Composés de l'arsenic , Pharmacologie , Lignée cellulaire tumorale , ADN complémentaire , Analyse de profil d'expression de gènes , Régulation de l'expression des gènes tumoraux , Banque de gènes , Leucémie aiguë promyélocytaire , Anatomopathologie , Hybridation d'acides nucléiques , Méthodes , Séquençage par oligonucléotides en batterie , Oxydes , Pharmacologie , Analyse de séquence d'ADNRésumé
Objective To study the effect of-1 site single nucleotide polymorphism(SNP) on CCNH gene promoter transcription activity.Methods PCR and site-directed mutagenesis technology were used to construct CCNH basic promoter and-1G mutate promoter.Dual-Luciferase Reporter assay system was used to detect the transcription activity of constructed promoter.Results In AD293 cells,the activity of-1G mutate type promoter was significantly lower than that of wild type-1T promoter(P
Résumé
<p><b>OBJECTIVE</b>To explore the distribution of monoamine oxidase A (MOA-A) EcoRV polymorphism in Shanghai Han population and its possible role in the risk for Parkinson's disease(PD).</p><p><b>METHODS</b>The MAO-A gene EcoRV polymorphism was detected with PCR-RFLP method in 110 PD patients and 182 healthy controls, furthermore, statistical analysis was performed to investigate association between EcoR V polymorphism and PD onset.</p><p><b>RESULTS</b>(1)Remarkable difference in MAO-A EcoR V polymorphic distribution has been observed between Shanghai Han population and that in North America. (2) Neither allelic frequency nor genotypic frequency in PD cases differs significantly from that in healthy controls regardless of data from male or female subclass.</p><p><b>CONCLUSION</b>There may be racial difference in the distribution of the human MAO-A EcoR V (C/T) polymorphism, but the present research does not support the association between this variant and susceptibility to PD in Chinese Han population of Shanghai area.</p>