摘要目的 探讨可移植性人癌组织重构过程中,肿瘤干细胞植入方与宿主受体间的关系.方法 将红色荧光蛋白(RFP)基因稳定转染于已建株的胶质瘤干祖细胞SU3,再将建立的SU3-RFP细胞接种于表达绿色荧光蛋白(GFP)的裸小鼠脑内.取移植瘤组织,筛选高增殖力GFP细胞,单克隆化后,鉴定细胞类型,测定其DNA含量,行染色体显带分析.并将建系的GFP细胞H1和H9分别接种于裸小鼠腋下和颅内,观察致瘤率和癌性表型.结果 在可移植性肿瘤组织中,RFP、GFP和RFP/GFP细胞的比例分别为(88.99±1.46)％、(5.59±1.00)％和(4.11±1.02)％.经2次单克隆化的宿主GFP细胞株H1和H9,在体外表现出无限增殖、接触抑制消失、超四倍体等癌细胞特征,并表达少突胶质细胞特异性标志蛋白2’,3’-环腺苷酸-3’-磷酸二酯酶(CNP).H1和H2细胞的体内致瘤率均为100％,肿瘤均向接种周围组织呈广泛侵袭性生长.结论 在可移植性胶质瘤模型中的肿瘤细胞,并非像传统观点那样均由移植的肿瘤干祖细胞产生,被接种的肿瘤干细胞侵袭的宿主脑细胞也可以转化为肿瘤细胞.

abstractsObjective During the process of tissue remodeling in human tumor transplantation models,the roles of the inoculated tumor cells and host tissue in tumor progression is still largely unknown.The aim of this study was to investigate the relationships and interactions between these two sides using GFP-RFP double fluorescence tracing technique.Methods Red fluorescence protein (RFP) gene was stably transfected into glioma stem cell line SU3,then SU3-RFP cells were transplanted into the brain of athymic nude mice with green fluorescence protein (GFP) expression.After the intracerebral tumors were formed,the relationship and interaction between GFP cells and RFP cells were analyzed.Highly proliferative GFP cells were screened out,and monocloned with micro-pipetting.DNA content assay,chromosome banding and carcinogenicity test of the GFP cells were performed to observe the GFP cells'cancerous phenotype in nude mice.Results In the transplantable tumor tissue,besides a great quantity of RFP cells,there were still a proportion of GFP cells and GFP/RFP fusion cells.The proportion of RFP cells,GFP cells and GFP/RFP cells were (88.99 ± 1.46) ％,(5.59 ± 1.00) ％,and (4.11 ± 1.020) ％,respectively.Two monoclonal host GFP cells (H1 and H9) were cloned,which demonstrated the properties of immortality,loss of contact inhibition,and ultra-tetraploid when cultured in vitro.Both H1 and H9 cells expressed CNP,a specific marker of oligodendrocytes.The GFP cells also demonstrated 100％ tumorigenic rate and high invasive properties in vivo.Conclusions In this glioma transplantation model,the transplanted tumor tissues contained not only transplanted glioma stem cells but also cancerous host GFP cells.Our findings offer important clues to further research on the relationships among different members in the tumor microenvironment.