CAJ | 학술논문

目的建立稳定、高表达绿色荧光蛋白(GFP)标记的裸鼠结肠癌原位移植模型并观察其生物特性.方法构建GFP pLPCX逆转录病毒质粒,将其转染至人结肠癌HCT116细胞,建立裸鼠的皮下瘤模型,活体荧光影像观察到绿色荧光标志的肿瘤生长至10 mm×10 mm,取肿瘤分别对15只BALB/C裸鼠进行结肠原位移植.利用荧光影像系统在不同时间点观察裸鼠原位移植瘤的生长、转移情况.不同时间点的卡尺与荧光测量结果分析采用t检验,组间比较采用重复测量设计的方差分析.结果 15只裸鼠结肠癌原位荧光移植模型均成功构建,未发生与手术相关的并发症和死亡.术后第3周,所有动物模型在荧光下观察到肿瘤.肿瘤体积随着时间推移逐步增大,术后3、4、5、6、7周不同时间点整体荧光影像测量原位移植肿瘤计算的体积＞体表游标卡尺测量计算的体积,但差异无统计学意义(f=-1.280,-1.115,-0.718,-0.199,-0.386,P＞0.05)；测量方式与时间的交互作用,两种测量方法的结果比较,差异有统计学意义(F =29.546,P＜0.05).实验终点时存活8只动物模型,其中6/8的动物模型发生肿瘤转移.结论结肠癌荧光原位移植模型技术可行,能够进行体内实时及无创地动态观察和分析肿瘤细胞的生长与转移情况.
목적 건립은정、고표체록색형광단백(GFP)표기적라서결장암원위이식모형병관찰기생물특성.방법 구건GFP pLPCX역전록병독질립,장기전염지인결장암HCT116세포,건립라서적피하류모형,활체형광영상관찰도록색형광표지적종류생장지10 mm×10 mm,취종류분별대15지BALB/C라서진행결장원위이식.이용형광영상계통재불동시간점관찰라서원위이식류적생장、전이정황.불동시간점적잡척여형광측량결과분석채용t검험,조간비교채용중복측량설계적방차분석.결과 15지라서결장암원위형광이식모형균성공구건,미발생여수술상관적병발증화사망.술후제3주,소유동물모형재형광하관찰도종류.종류체적수착시간추이축보증대,술후3、4、5、6、7주불동시간점정체형광영상측량원위이식종류계산적체적＞체표유표잡척측량계산적체적,단차이무통계학의의(f=-1.280,-1.115,-0.718,-0.199,-0.386,P＞0.05)；측량방식여시간적교호작용,량충측량방법적결과비교,차이유통계학의의(F =29.546,P＜0.05).실험종점시존활8지동물모형,기중6/8적동물모형발생종류전이.결론 결장암형광원위이식모형기술가행,능구진행체내실시급무창지동태관찰화분석종류세포적생장여전이정황.
Objective To establish a stable orthotopic model with high green fluorescent protein (GFP) expression in nude mice and observe its biological features.Methods Human HCT116 colon cancer cells transfected with GFP pLPCX retroviral plasmid were used to build a subcutaneous tumor model in nude mice.Fifteen BALB/C nude mice were selected to underwent orthotopic transplantation of colon when the GFP-labeled tumor grew to 10 mm × 10 mm as observed by in vivo fluorescent microscopy.The growth and metastasis of orthotopically implanted colon cancer cells were observed with fluorescent imaging system at different time points.The differences of the tumor size measured by peripheral vernier caliper and fluorescent imaging system were analyzed using the t test,and the differences in different groups were analyzed using the analysis of variance.Results GFP-labeled colon cancer models were successfully established in all the 15 nude mice,and there was no surgery-related complications or death.Tumors marked by GFP were observed under fluoroscope in week 3.The size of the tumors progressively increased with time.The volumes of the orthotopically transplanted tumors obtained from global measurement using fluorescent imaging system were greater than those measured by peripheral vernier calipers at postoperative week 3,4,5,6,7,while no statistically significant difference was observed (t =-1.280,-1.115,-0.718,-0.199,-0.386,P ＞0.05).There was a significant difference in the interation of measure method and different time points (F =29.546,P ＜ 0.05).Eight nude mice survived at the end of the experiment,and tumor metastasis was observed in 6 mice.Conclusions It is technically feasible to construct GFP-labeled colon cancer orthotopic transplantation model.The mice model could be used for real-time,in vivo,non-invasive and dynamic observation and analysis of the growth and metastasis of tumor cells.