CAJ | 학술논문

目的：探讨绿色荧光蛋白（ GFP）标记的大鼠C6胶质瘤模型的构建方法并观察其作用。方法用带有增强型GFP（ EGFP）基因的pEGFP-C1质粒转染C6细胞，通过G418筛选获得稳定表达EGFP的C6细胞克隆EGFP-C1-C6细胞。采用立体定向法，将浓度为1×106/L的EGFP-C1-C6细胞悬液10μL注入16只SD大鼠的颅内，建立EGFP标记的大鼠C6胶质瘤模型。观察大鼠接种肿瘤细胞后的反应及存活时间。分别于造模后第7、14、21天对大鼠行头颅MRI检查，计算肿瘤体积。常规HE染色观察病理学形态，激光共聚焦显微镜下观察胶质瘤的浸润及侵袭情况。结果成功培养了具有EGFP标记的EGFP-C1-C6细胞，MRI及病理学检查发现所有大鼠颅内均有肿瘤生长，肿瘤体积为（244．60±36．06） mm3，采用激光共聚焦显微镜观察易于发现经EGFP标记的肿瘤组织，肿瘤区域清晰可见，并较易发现单个肿瘤细胞的远处浸润生长。结论采用EGFP-C1-C6细胞成功构建了大鼠C6胶质瘤模型，该模型有助于显示胶质瘤细胞的侵袭、转移及微小侵袭灶。
목적：탐토록색형광단백（ GFP）표기적대서C6효질류모형적구건방법병관찰기작용。방법용대유증강형GFP（ EGFP）기인적pEGFP-C1질립전염C6세포，통과G418사선획득은정표체EGFP적C6세포극륭EGFP-C1-C6세포。채용입체정향법，장농도위1×106/L적EGFP-C1-C6세포현액10μL주입16지SD대서적로내，건립EGFP표기적대서C6효질류모형。관찰대서접충종류세포후적반응급존활시간。분별우조모후제7、14、21천대대서행두로MRI검사，계산종류체적。상규HE염색관찰병이학형태，격광공취초현미경하관찰효질류적침윤급침습정황。결과성공배양료구유EGFP표기적EGFP-C1-C6세포，MRI급병이학검사발현소유대서로내균유종류생장，종류체적위（244．60±36．06） mm3，채용격광공취초현미경관찰역우발현경EGFP표기적종류조직，종류구역청석가견，병교역발현단개종류세포적원처침윤생장。결론채용EGFP-C1-C6세포성공구건료대서C6효질류모형，해모형유조우현시효질류세포적침습、전이급미소침습조。
Objective To establish the rat C6 glioma model engineered by green fluorescent protein and explore its effect.Methods C6 cells were transfected with pEGFP-C1 plasmid expressing enhanced green fluorescent protein ( EG-FP) gene.Then the transfected cells were selected by G418 to obtain the stable C6 cell clones ( pEGFP-C1-C6) .pEGFP-C1-C6 cell suspension was injected into intracal of 16 SD rats to establish rat C6 glioma model with green fluorescent protein by stereotaxic apparatus.After inoculation, cell life cycle was observed and tumor growth status was dynamically documen-ted by magnetic resonance imaging.At 7, 14, 21 day after glioma model established, glioma tumor volumes were detected and measured by MRI.Brain specimens were observed by HE staining.Confocal laser scanning microscopy was used to ob-serve the infiltration and invasion of glioma cell.Results Rat C6 glioma model with green fluorescent protein was estab-lished successfully.Tumor cells invasion can be easily detected by green fluorescent protein imaging and MRI.Tumor vol-ume was (244.60 ±36.06)mm3.It was easier to observe the micro-metastasis and invasive.Conclusions Rat C6 glioma model could be successful established with EGFP-C1-C6 cells, which was more sensitive in showing invade, metastasis and single invasive tumor cell.