论文部分内容阅读
目的建立稳定自发绿色荧光的高、低转移人肝癌细胞株并明确鉴定。方法脂质体转染法将绿色荧光蛋白(GFP)基因分别转染至高、低转移人肝癌细胞株MHCC97-H／L内,经 G418反复筛选并单克隆化,得到两株新细胞株MHCC97-HG／LG,对新细胞株GFP表达稳定性、生物学特性、体内外侵袭转移能力及肿瘤血管生成等进行鉴定。结果 MHCC97-HG／LG在体外培养或裸鼠体内接种36 d后均能稳定表达绿色荧光。MHCC97-HG肝、肺转移率均为100％,较 MHCC97-LG及其父系体内外侵袭转移能力显著增强(P<0．01),其原位肿瘤微血管密度较 MHCC97-LG显著增加[(121．0±15．9)／HP vs(87．0±14．2)／HP,P<0．01]。体视荧光显微镜可实时观察裸鼠体内MHCC97-HG／LG种植瘤生长、转移及血管形成情况。结论 MHCC97-HG／LG及其动物模型在肝癌转移机制、肿瘤血管生成等诸多研究领域可能有较广阔的应用前景。
Objective To establish a stable and spontaneous green fluorescence of high and low metastatic human hepatoma cell lines and clear identification. Methods The green fluorescent protein (GFP) gene was transfected into human hepatocellular carcinoma cell line MHCC97-H / L by lipofection method. The recombinant plasmid was repeatedly screened and cloned by G418 to obtain two new cell lines, MHCC97- HG / LG, the new cell line GFP expression stability, biological characteristics, in vitro invasion and metastasis and tumor angiogenesis were identified. Results MHCC97-HG / LG stably expressed green fluorescence after 36 days of in vitro culture or nude mice inoculation. The MHCC97-HG liver and lung metastasis rates were 100%, significantly higher than that of MHCC97-LG and its paternal offspring (P <0.01), and the in situ tumor microvessel density was significantly higher than that of MHCC97-LG [(121 .0 ± 15.9) / HP vs (87.0 ± 14.2) / HP, P <0.01]. The fluorescence microscope can observe the growth, metastasis and angiogenesis of MHCC97-HG / LG implanted tumor in nude mice in real time. Conclusion MHCC97-HG / LG and its animal model may have broad application prospects in many fields such as metastasis of liver cancer and tumor angiogenesis.