ABSTRACT
Humanized monoclonal antibodies(mAbs) are increasingly widely used in targeted therapy for cancer and some other major diseases.Complementarity-determining region(CDR) grafting makes quantities of humanized mAbs available.Herein,we provide an overview on the strategy and progress of CDR grafting.
ABSTRACT
Objective To study the activity of the survivin gene promoter in several tumor cell lines and evaluate the possible application of this promoter in tumor gene therapy. Methods ①The expressions of survivin gene in A549,MDA-MB231 and HepG2 cell lines were detected by RT-PCR and Western blotting.②Tumor cells(A549,MDA-MB231,HepG2) were transiently transfected by reporter plasmids containing different length of survivin promoter using lipofectamine.And 48 h later,the level of reporter gene expression was analyzed.Results There were different levels of survivin expression in A549,MDA-MB231 and HepG2 cell lines.Transient transfection assay approved that pLuc-surP-987,pLuc-surP-596,pLuc-surP-269 and pLuc-surP-158 showed high activity and 269 bp survivin promoter demonstrated the highest activity. Conclusion In transcriptional level,survivin promoter can activate the reporter gene in several tumor cell lines.It is a potential candidate promoter in tumor gene therapy.
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Objective To upregulate Notch signaling in cancer cells by overexpression of active part of Notch1 and to examine the proliferation of the cells. Methods Four cancer cell lines were infected with retrovirus recombined with sequence encoding active part of Notch1.CBF-1 reporter plasmid was used to detect Notch signaling and proliferation assay was carried out by MTS method.Cell cycle analysis was synchronously conducted. Results The overexpression of the active part of Notch1 induced upregulation of Notch signaling,led to growth inhibition in Hela and HepG2 cell lines and growth boost in BGC-823 cell lines,while had no effect on Chang cell lines. Conclusion The upregulation of Notch signaling can exert various effects on different cancer cell lines which is critical to the gene therapy for cancers.
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Objective To construct luciferase reporter plasmid containing synthetic CArG elements and to investigate their radiation-inducible property in tumor cells. Methods Insert chimeric regulation elements consisting of nine tandem-repeat copies of CArG sequence (CCATATAAGG) and CMV IE basal gene promoters into pGL3-Basic vectors to construct luciferase reporter plasmids. Tumor cells(HeLa, A549 and HepG2) were transiently transfected by reporter plasmids using lipofectamine, and transfected cells were irradiated by ?-ray with different doses. After 36 h, we assayed the level of reporter gene expression. Results The CArG elements could successfully induce the expression of a downstream reporter gene following irradiation, with maximal expression seen after 3 Gy irradiation. Conclusion The synthetic CArG elements are responsive to low dose of radiation and are able to enhance down-stream gene expression. It is expected to be the essential potential for future application of radiogenetic cancer therapy.