CAJ | 학술논문

目的在已报道的原位移植的胶质瘤间质细胞可发生恶性转化的基础上,进一步探索该转化的发生是否依赖肿瘤部位特异性微环境.方法将转有红色荧光蛋白(RFP)基因的人胶质瘤干/祖细胞SU3-RFP接种于全身表达绿色荧光蛋白(GFP)的裸小鼠肝脏,致瘤后取移植瘤组织做常规病理检测和细胞培养,单克隆具永生化的GFP+,GFP+/RFP+细胞传代,做细胞表型分析和致瘤实验.结果 SU3-RFP的移植成功率约83％,经异位生长的移植瘤组织再培养获得永生化的间质细胞.已建立B4,B9,B10三株单克隆细胞株,其中BB4为共表达GFP/RFP的融合细胞,还同时表达CD1a、CD83、CD86等树突状细胞标志蛋白;B9表达GFP和巨噬细胞标志蛋白CD68、F4/80;B10表达GFP和成纤维细胞标志蛋白FAP-α、α-SMA、S100均为鼠源宿主细胞.3株细胞遗传学检测都是异倍体,在裸小鼠皮下致瘤率均为100％.结论在裸小鼠肝组织内建立的胶质瘤异位移植模型中,胶质瘤间质细胞也能恶性转化,表明这种恶性转化在中枢神经系统之外亦能发生,而且由间质细胞恶性转化可引起肿瘤异质性,这对进一步研究肿瘤发生、演进与肿瘤微环境的关系具有重要意义.
목적 재이보도적원위이식적효질류간질세포가발생악성전화적기출상,진일보탐색해전화적발생시부의뢰종류부위특이성미배경.방법 장전유홍색형광단백(RFP)기인적인효질류간/조세포SU3-RFP접충우전신표체록색형광단백(GFP)적라소서간장,치류후취이식류조직주상규병리검측화세포배양,단극륭구영생화적GFP+,GFP+/RFP+세포전대,주세포표형분석화치류실험.결과 SU3-RFP적이식성공솔약83％,경이위생장적이식류조직재배양획득영생화적간질세포.이건립B4,B9,B10삼주단극륭세포주,기중BB4위공표체GFP/RFP적융합세포,환동시표체CD1a、CD83、CD86등수돌상세포표지단백;B9표체GFP화거서세포표지단백CD68、F4/80;B10표체GFP화성섬유세포표지단백FAP-α、α-SMA、S100균위서원숙주세포.3주세포유전학검측도시이배체,재라소서피하치류솔균위100％.결론 재라소서간조직내건립적효질류이위이식모형중,효질류간질세포야능악성전화,표명저충악성전화재중추신경계통지외역능발생,이차유간질세포악성전화가인기종류이질성,저대진일보연구종류발생、연진여종류미배경적관계구유중요의의.
Objective Tumor stromal cells have the potential of undergoing malignant transformation induced by glioma stem cells (GSCs) in orthotopic glioma model.The purpose of this study was to explore whether malignant transformation of tumor stromal cells induced by GSCs is dependent on specific local microenvironment.Methods Human glioma stem/progenitor cell line SU3 transfected with red fluorescent protein (SU3-RFP) gene were implanted into the liver of nude mice with whole-body expressing green fluorescence protein (GFP).Then hepatic tumors were harvested to prepare single cell suspension and analyzed with routine pathological examinations.GFP cells with high proliferative abilities were obtained from the cultivation of single cell suspension.Immortalized glioma stromal cells only expressing GFP and double expressing GFP/RFP were further monocloned with micro-pipetting techniques and under continuous passages.Cell phenotypic analysis and tumorigenicity tests were also performed.Results SU3-RFP was transplanted into liver with a tumor formation rate of 83％.Immortalized glioma stromal cells were obtained from re-cultured xenograft tumor tissue.Three monoclonal cell lines B4,B9,B10 were established and proved to be host-derived cells.B4 was found to be a fusion cell co-expressing GFP/RFP and dendritic cell markers CD1a,CD83 and CD86.Both B9 and B10 were GFP+ cells.B9 expressed macrophage markers CD68 and F4/80 while B10 produced fibroblast marker proteins FAP-α,α-SMA and S100.Three cells were all aneuploid with a tumorigenicity rate of 100％ in nude mice.Conclusion Tumor stromal cells have the potential of malignant transformation in a heterotopic xenograft glioma model.Malignant transformation may also occur outside the central nervous system and contribute to tumor heterogeneity.Further studies are warranted for elucidating the relationship between tumorigenesis,evolution and tumor microenvironment.