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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 mLumin + HepG2 cell line, which will lay the foundation for the further establishment of tumor model and small animal fluorescence imaging. Methods: The pcDNA3.0-mLumin plasmid was transfected into HepG2 cells by calcium phosphate method. After screening by G418, the expression of mLumin gene in HepG2 cells was detected by fluorescence microscope. The expression of mLumin gene in HepG2 cells was detected by fluorescence microscopy And flow cytometry detection of mLumin + HepG2 cell ratio, and finally inoculated in nude mice subcutaneously to observe tumorigenicity. Results: The mRNA expression of mLumin gene in HepG2 cells was confirmed by RT-PCR. Under the inverted fluorescence microscope, when using green light (540 nm) excitation, you can see a large number of red fluorescent cells, flow cytometry positive rate of 50%. Small animal live imager showed that the nude mice were subcutaneously inoculated into the tumor, frozen sections of the tissue also confirmed a large number of HepG2 cells in mLumin protein expression. Conclusion: The successful establishment of mLumin + HepG2 cells laid the foundation for the further study of tumor.