ABSTRACT
OBJECTIVE: To explore the possible molecular mechanism of Ikaros regulation on FUT4 expression by analyzing the correlation of the functional state of Ikaros with level of FUT4 expression, so as to provide the theoretical basis for personalized treatment in children with ALL. METHODS: The subtypes of Ikaros were identified by nested PCR and sequencing. The expression level of FUT4 was detected by quantitative PCR and analyzed by ΔΔCt method in the early stage of treatment, remission and relapse of ALL. RESULTS: Ik1 and Ik2 were the main functional subtypes, and the dominant negative Ikaros was Ik6; the Ik6 was detected in 23 patients with ALL. It was found that 2.73% patients expressing Ik6 alone and 18.18% patients with heterozygous expression were detected. The expression of FUT4 in the newly diagnosed ALL was higher than that in the control group, and the functional Ikaros negatively correlated with the FUT4 expression(r=-0.6329). CONCLUSION: Dominant negative Ikaros closely correlated with the relapse of acute lymphoblastic leukemia in children. The functional Ikaros negatively correlated with FUT4 expression. Ikaros inhibit the transcriptional activity of FUT4, that may be the molecular mechanism of Ikaros regulating the expression of FUT4.
Subject(s)
Fucosyltransferases/metabolism , Ikaros Transcription Factor/metabolism , Lewis X Antigen/metabolism , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Acute Disease , Child , Humans , Protein Isoforms , RecurrenceABSTRACT
The phytochemical progress on Angelica sinensis (Oliv.) Diels over the past decades is summarized. Since 1970s, 165 chemical constituents, including phthalides, phenylpropanoids, terpenoids and essential oils, aromatic compounds, alkaloids, alkynes, sterols, fatty acids, and polysaccharides have been isolated or detected from the various parts of the title plant.
Subject(s)
Angelica sinensis/chemistry , Phytochemicals/isolation & purification , Alkaloids/isolation & purification , Alkynes/isolation & purification , Benzofurans/isolation & purification , Fatty Acids/isolation & purification , Oils, Volatile/isolation & purification , Phytosterols/isolation & purification , Polysaccharides/isolation & purification , Propanols/isolation & purification , Terpenes/isolation & purificationABSTRACT
OBJECTIVE: To prepare anti-CD3 and anti-Igm&mgr; chain bispecific antibody (BsAb) by means of cell fusion and assess the stability and activity of the hybrid hybridoma. METHODS: Mouse hybridoma cell line secreting anti-human CD3 monoclonal antibody (mAb) was trans formed into HAT-sensitive cells by 8-azaguanine, which was subsequently transfected by plasmid pCDaA3 containing neoR marker gene via FuGENETM6. The resulted mutant phenotype, named alphaCD3 HATs G418R, was fused with the hybridoma producing anti-IgM&mgr; chain mAb, and enzyme-linked immunosorbent assay and flow cytometry were employed to identify the fusion cells producing the target BsAb. RESULTS: Six rounds of cell fusion was performed and a total of l 080 wells inoculated, yielding 5 hybrid hybridoma cell lines. Two of the 5 cell lines were subcloned and continuously cultured in vitro for 2 months without losing their capability to secrete BsAb. CONCLUSION: Cell fusion technique can be utilized to prepare BsAb-producing hybrid hybridoma that has similar stability and activity to its parent hybridoma.