电穿孔法转染GFP标记H22和S180的实验研究

采用电转染法将绿色荧光蛋白(GFP)基因导入小鼠肝癌H22和肉瘤S180细胞,通过G418筛选,建立了稳定表达该蛋白的小鼠肝癌H22细胞株和肉瘤S180细胞株;光镜和电镜观察其细胞形态、超微结构及生长状况没有发生显著改变;建立相应的皮下和腹腔荷瘤动物模型,观察到其皮下和腹腔成瘤时间与腹腔荷瘤生存时间无显著改变(P＞0.05),在活体荧光成像系统能观察到其荷瘤部位荧光.期望利用gfpH22和gfpS180阳性细胞株,运用免疫荧光技术、活体荧光成像系统和激光共聚焦系统对肿瘤进行体内外直观、可视和半定量的深入研究.

Research of GFP-expressing H22 Cells and S180 Cells by Electroblot in Vitro and in Vivo

Mouse hepatoma H22 cells and sarcoma cells(H22 and S180) were infected with EGFP-N1 by electroblot,and the acquired gfp-H22 and gfp-S180 cells expressing strong green fluorescence protein(GFP) fluorescence were supplemented with medium G418 Sigma(800mg/ml).Meanwhile,the models bearing cancer(gfp H22 and gfp S180) subcutaneously and with abdominal cavity were established.There were no statistically significant differences by comparison on the cell phenotype,ultramicrostructure,growth curve and bearing cancer time between the H22 cells and S180 cells(P>0.05).The GFP fluorescence was detected with whole body GFP imaging system in vivo and with fluorescence microscope.According to the results of in vitro and in vivo assay,it was shown that,by application of fluorescence technology,the GFP-expressing H22 cells and S180 cells could be used in further studies on the tumor biological behavior.