CD44、CD133和ABCG2潜在癌症生物标记蛋白质的表达(英文)

Expression of CD44,CD133,and ABCG2 as Potential Cancer Biomarker Proteins

癌症生物标记物是一种新的早期癌症治疗工具,它既可用于癌细胞抑制,也可用于将其杀灭.癌干细胞已被证明是癌症的一个关键标志细胞.最近,随着单克隆抗体技术的发展生物标记物的研究发展迅速,一些重要的癌干细胞标记已被确认.单克隆抗体是癌标记物应用的重要技术,也是临床诊断和治疗重要手段.本研究的癌干细胞标记物CD44,CD133和ABCG2,分别通过Genestar等软件分析,并选定目标抗原特异性的基板的基因,设计出目标基因的模板和引物.经过免疫分析,设计出编码基因,利用PCR克隆技术,将其并插入预先选定的pGEX-4T-1或pET32a质粒上的克隆位点上.载有预定基因的标记物,通过载体转入感受态的E.coli BL21中,在加入IPTG的条件下,诱导其蛋白质的表达,得到目标蛋白质产物。利用SDS-PAGE和Western Blot analysis,验证其表达的正确性.研究的目标蛋白质片段CD44、CD133和ABCG2,可以用于癌干细胞进一步的研究和其单克隆抗体的制作.

Cancer biomarkers can provide the chance to develop therapies for cancer during its the natural progression,to either inhibit or eliminate the disease.Cancer stem cells have been proved to be critical in the progression of cancer.Recently,some important cancer stem cells markers have been characterized.The rapid development of cancer biomarker research is attributed to a large extent to the advance of the monoclonal antibody technology.Monoclonal antibodies specific for cancer biomarkers have important clinical use in diagnosis and therapeutics.In the present studies,the cancer stem cell markers,CD44,CD133 and ABCG2,were first analyzed for their respective antigenic epitopes by Genestar.Then the gene encoding protein fragment with sound immunogenicity was cloned by PCR and inserted to the multiple cloning site of the pGEX-4T-1 or pET32a plasmid.The resulting expression vector was transformed into the E.coli BL21 strain and the protein expression was induced by the addition of IPTG.Finally,the target protein was characterized by the SDS-PAGE and Western Blot analysis.In further study,these CD44,CD133 and ABCG2 target protein fragments will be utilized to develop monoclonal antibodies,which can be used in cancer stem cell research.