CAJ | 학술논문

目的建立具有器官转移亲嗜性的单克隆人肝癌细胞株和相应的裸鼠移植模型. 方法取肺和淋巴结转移亲嗜性人肝癌荧光细胞株HCCLM3-R-LM1及HCCLM3-R-LnM1,通过极限稀释法进行单细胞培养,获得8个HCCLM3-R-LM1来源的单克隆细胞株(LM1-S2,-S3,-S4,-S5,-S11,-S15,-S21,-S23)和5个HCCLM3-R-LnM1来源的单克隆细胞株(LnM1-S7,-S11,-S13,-S17,-S20);将上述人肝癌单克隆细胞分别接种于4周龄的裸鼠皮下,3周后皮下瘤组织移植至裸鼠的肝脏,6周后观察裸鼠肺和腹腔淋巴结转移灶荧光面积,并与肺组织连续切片中的转移灶数目进行比较.同一株细胞的肺和淋巴结转移情况比较用Wilcoxon秩和检验,单克隆细胞株之间的肺、淋巴结转移情况比较用Kruskal-Wallis检验. 结果在13株人肝癌单克隆细胞中,有6株细胞形成皮下瘤;移植至裸鼠肝脏后表现出不同的转移潜能和器官靶向特性.其中LM1-S3,LM1-S4,LM1-S5和LM1-S11单克隆细胞的肺转移积分吸光度值分别为80923±10162、1506000±297064、36 140±8210和508448±1342729(P＜0.01),但不发生淋巴结转移.LnM1-S11单克隆细胞的肺与淋巴结转移灶积分吸光度值分别为435 062±206 620和1254000±225171. 结论成功建成了不同转移潜能和器官亲嗜性的人肝癌单克隆细胞株和裸鼠移植模型,其中LM1-S3,LM1-S4,LM1-S5和LM1-S11为肺特异亲嗜性的人肝癌单克隆细胞株,而LnM1-S11细胞为肺和淋巴结双重亲嗜性的人肝癌单克隆细胞株,为肝癌转移器官靶向性研究提供了理想的体内外模型.
목적 건립구유기관전이친기성적단극륭인간암세포주화상응적라서이식모형. 방법 취폐화림파결전이친기성인간암형광세포주HCCLM3-R-LM1급HCCLM3-R-LnM1,통과겁한희석법진행단세포배양,획득8개HCCLM3-R-LM1래원적단극륭세포주(LM1-S2,-S3,-S4,-S5,-S11,-S15,-S21,-S23)화5개HCCLM3-R-LnM1래원적단극륭세포주(LnM1-S7,-S11,-S13,-S17,-S20);장상술인간암단극륭세포분별접충우4주령적라서피하,3주후피하류조직이식지라서적간장,6주후관찰라서폐화복강림파결전이조형광면적,병여폐조직련속절편중적전이조수목진행비교.동일주세포적폐화림파결전이정황비교용Wilcoxon질화검험,단극륭세포주지간적폐、림파결전이정황비교용Kruskal-Wallis검험. 결과 재13주인간암단극륭세포중,유6주세포형성피하류;이식지라서간장후표현출불동적전이잠능화기관파향특성.기중LM1-S3,LM1-S4,LM1-S5화LM1-S11단극륭세포적폐전이적분흡광도치분별위80923±10162、1506000±297064、36 140±8210화508448±1342729(P＜0.01),단불발생림파결전이.LnM1-S11단극륭세포적폐여림파결전이조적분흡광도치분별위435 062±206 620화1254000±225171. 결론 성공건성료불동전이잠능화기관친기성적인간암단극륭세포주화라서이식모형,기중LM1-S3,LM1-S4,LM1-S5화LM1-S11위폐특이친기성적인간암단극륭세포주,이LnM1-S11세포위폐화림파결쌍중친기성적인간암단극륭세포주,위간암전이기관파향성연구제공료이상적체내외모형.
Objective To establish a single cell-derived organ site-specific metastatic model of human hepatocellular carcinoma (HCC) in the nude mouse.Methods Using the limited dilution method,HCCLM3-R-LM1 and HCCLM3-R-LnM1 cell lines were used to genrerate eight (LM1-S2,-S3,-S4,-S5,-S11,-S15,-S21,and -S23) and five (LnM1-S7,-S11,-S13,-S17,and -S20) single cell-derived monoclonal cell lines,respectively.The monoclonal cell lines were seeded into 4-week-old nude mice,and three weeks later the resultant subcutaneous tumor tissues were orthotopically transplanted into the livers of nude mice.At six weeks after implantation,lung and lymph node were extracted for analysis of the metastatic foci fluorescence area and pathology to assess the number of metastatic foci.Results Among the 13 mice implanted with the established monoclonal cell lines,six grew subcutaneous tumors.When orthotopically transplanted,the six tumors showed remarkably different metastatic potential and organ site-specific tropism.The fluorescence areas of lung metastatic foci were:LM1-S3,80923± 10162; LM1-S4,1506000±297064;LM1-S5,36140±8210; and LM1-S11,508448± 134272 (P＜0.01); no lymph node metastases were found for these lines.For LnM1-S11,the fluorescence areas of lung and lymph node metastatic foci were 435 062±206620 and 1 254000±225 171,respectively.Conclusion We successfully established several monoclonal cell lines and nude mouse models of HCC with different metastatic potential and organ tropism.Among them,LM1-S3,LM1-S4,LM1-SS,and LM1-S11 have metastasis organotropism to lung.The LnM1-S11 line exhibits dual metastasis organotropism to lung and lymph node.These monoclonal cell lines and nude mouse models may represent useful tools for study of HCC metastasis organotropism.