mLumin转染人肝癌HepG2细胞系的建立  

Establishment of mLumin+ human hepatoma carcinoma cell line HepG2 transfected by pcDNA3.0-mLumin

作　　者：刘霞[1] 闫美娜[1] 王荟[1] 万洋洋[1] 许凤姣[1] 邵青[1] 成静[1] 范兴文[1] 石慧娜[1] 邵启祥[1] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2011年第4期322-325,F0003,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(30671984);江苏省自然科学基金资助项目(BK2008231);江苏省研究生创新计划项目(CX10B_283Z);江苏大学大学生科研立项项目(09A075)

摘　　要：目的:建立mLumin+的HepG2细胞系,为进一步建立肿瘤模型和小动物荧光成像观察奠定基础。方法:采用磷酸钙法,将pcDNA3.0-mLumin质粒转染至人肝癌细胞系HepG2,经G418筛选扩增后通过PCR检测mLumin基因在转染的肝癌细胞HepG2中的表达情况,同时应用荧光显微镜及流式细胞仪检测mLumin+的HepG2细胞比例,最终将其接种于裸鼠的皮下,观察致瘤情况。结果:RT-PCR鉴定证实肝癌细胞HepG2中有mLumin基因的mRNA表达。在倒置荧光显微镜下,采用绿光(540 nm)激发时,可以见到大量的发射红色荧光的细胞,流式细胞术检测有50%的阳性率。小动物活体成像仪显示裸鼠皮下接种能够成瘤,组织冰冻切片也证实HepG2细胞中有大量mLumin蛋白表达。结论:成功建立了mLumin+的HepG2细胞,为进一步进行肿瘤的基础和应用研究奠定了基础。Objective： To establish the mLumin＋ HepG2 cell line for the further development of animal tumor models and scanning the fluorescence image of the animal under the small animal fluorescence imager.Methods： The recombinant vectors containning mLumin and neomycin resistance gene（neo）were transduced into HepG2 cells by the calcium phosphate method.The neo resistant cells were selected by G418 for two weeks.The mRNA expression of mLumin protein in HepG2 cells was confirmed by RT-PCR.Red fluorescent from the mLumin protein was observed under the fluorescence microscope and the red fluorescent mLumin protein positive cells were detected by flow cytometer（FCM）.The tumor genesis abilities of the mLumin＋HepG2 cells were validated by a nude mice tumor model.Results： The mRNA of mLumin was expressed in high level in HepG2 cells.The transfected HepG2 cells showed as the red fluorescence mLumin positive cells under the inverted fluorescence microscope.There were about 50% mLumin＋ HepG2 cells detected by FCM.Furthermore,in-vivo small animal imaging showed that the tumors were growth up with different concentrations mLumin＋HepG2 cells.Conclusion： The mLumin＋HepG2 cells have been establised successfully and can be used for the further research about the tumor.

关 键 词：mLumin基因 HEPG2 活体成像 

分 类 号：R392.9[医药卫生—免疫学]