荧光成像技术活体示踪裸鼠肝细胞癌肺转移  

Tracking pulmonary metastasis of hepatocellular carcinoma of nude mouse using fluorescence imaging

作　　者：叶志强[1] 刘旭辉[1] 吴飞龙[1] 赵坤[2] 潘卫东[2] 

机构地区：[1]中山大学附属第三医院急诊科,广州510630 [2]中山大学附属第三医院肝胆外科,广州510630

出　　处：《中华肝脏外科手术学电子杂志》2014年第5期43-46,共4页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：国家自然科学基金(81172783)

摘　　要：目的探讨荧光素酶标记肝细胞癌（肝癌）细胞的荧光成像技术在活体示踪裸鼠肝癌肺转移中的价值。方法应用低转移潜能 HCC97L、高转移潜能HCC97H 肝癌细胞建立稳定表达荧光素酶的肝癌细胞株 HCC97L-FL、HCC97H-FL。24只裸鼠按随机数字表法随机分为HCC97L-FL 组和 HCC97H-FL 组。分别将5＆#215;106个 HCC97L-FL 和 HCC97H-FL 细胞加入磷酸盐缓冲液（PBS）50μl，制成细胞悬液。将细胞悬液注入裸鼠尾静脉，建立荧光素示踪的裸鼠肝癌肺转移模型。观察荧光成像示踪肺转移瘤大体标本及组织标本情况。两组数据比较采用 t 检验。结果荧光检测发现，HCC97L-FL、HCC97H-FL 细胞均可发出强烈荧光。建模后2～3周，HCC97H-FL 组裸鼠的肺部收集到荧光信号，3～4周荧光强度逐渐增强；HCC97L-FL 组裸鼠无可见的肺部荧光信号。建模后4周 HCC97H-FL 组荧光强度为（1.73±0.16）×10^4 a.u.，明显高于 HCC97L-FL 组的（0.03±0.01）×10^4 a.u.（t=23.652, P〈0.05）。建模后4周 HCC97H-FL 组裸鼠肺脏密布大小不等转移瘤病灶，部分较大病灶突出于肺脏表面；HCC97L-FL 组中未观察到转移瘤。转移瘤癌细胞核异型性明显，具备肝癌特征。结论荧光素酶标记肝癌细胞的荧光成像技术可对裸鼠肝癌肺转移进行活体、动态示踪。该技术为体内肝癌细胞生长、侵袭、转移的研究提供了可靠的量化研究手段。Objective To investigate the value of fluorescence imaging of labeling the hepatocellular carcinoma （HCC） cells with luciferase in tracking the pulmonary metastasis of HCC of nude mouse . Methods Low metastatic potential HCC97L and high metastatic potential HCC97H cells were transfected into HCC cell lines HCC97L-FL, HCC97H-FL with stable expression of luciferase. A total of 24 nude mice were randomly divided into HCC97L-FL group and HCC97H-FL group by the random number table method. Cell suspension was prepared by adding HCC97L-FL, HCC97H-FL cells （5＆#215;106） into the phosphate buffered saline （PBS, 50 μl） respectively. Cell suspension was injected into the caudal vein of nude mice. The model of tracking pulmonary metastasis of HCC with fluorescein was established. The gross and tissue samples of the pulmonary metastasis tracked by fluorescence imaging were observed.The data between two groups were compared using t test. Results Through fluorescence detection, intensive fluorescence could be observed in the HCC97L-FL, HCC97H-FL cells. After 2 to 3 weeks of modeling, fluorescence signal was collected from the nude mice lung in HCC97L-FL group. The fluorescence intensity enhanced gradually in 3-4 week. No fluorescence signal was observed in the nude mice lung in HCC97H-FL group. After 4 weeks of modeling, the fluorescence intensity was （1.73 ± 0.16） ×10^4 a.u. in HCC97H-FL group, which was significantly higher than that in HCC97L-FL group [（0.03 ±0.01） ×10^4 a.u.] （t=23.652, P〈0.05）. The nude mice lung in HCC97H-FL group was densely covered with metastatic tumors of varying sizes after 4 weeks of modeling. Some of the bigger tumors were visible on the lung surface. No metastatic tumor was observed in the HCC97L-FL group. The metastatic tumor was observed with an obvious cell nuclear atypia and characteristic feature of HCC. Conclusions Fluorescence imaging of using luciferase to label HCC cells can dynamically track the pulmonary metastasis of HCC of nude mouse . This technique pr

关 键 词：癌 肝细胞 肝肿瘤 实验性 小鼠 裸 荧光素酶类 细菌 活体成像 活体示踪 肿瘤转移 

分 类 号：R735.7[医药卫生—肿瘤]