CAJ | 학술논문

目的克隆具有肺和淋巴结转移亲嗜性的人肝癌细胞并建立相应的裸鼠移植模型.方法将荧光人肝癌细胞HCCLM3-R接种于4周龄的裸鼠肝脏,6周后在荧光解剖下观察并获取肺及腹腔淋巴结转移灶组织,经体外克隆培养,所获细胞分别标记为HCCLM3-R-LM1和HCCLM3-R-LnM1.接种上述两种细胞于4周龄裸鼠肝脏,观察各自肺和腹腔淋巴结转移灶荧光数量,并与肺组织连续切片中的转移灶数目进行比较.计量资料采用Wilcoxon秩和检验和Kruskal-Wallis秩和检验进行统计学分析.结果 HCCLM3-R-LM1、HCCLM3-R和HCCLM3-R-LnM1细胞接种于裸鼠肝脏后第6周,在肺和腹腔淋巴结中均能发现肿瘤转移.3株细胞接种后,裸鼠肺和腹腔淋巴结转移灶荧光面积分别为8687.00±1844.63和2570.00±318.20(P＜0.001),6457.67±832.62和10 994.33±2212.31(P＜0.001),2968.67±2571.00和24 416.00±7186.13(P＜0.001),每只裸鼠光镜下肺转移灶中位数分别为755、430、310个(P＜0.001),与荧光定量结果相吻合.结论成功建成人肝癌肺和淋巴结亲嗜性转移细胞和裸鼠移植模型,其中HCCLM3-R-LM1细胞具有明显的肺转移特性,而HCCLM3-R-LnM1细胞具有明显的淋巴结转移特性,为肝癌器官靶向转移研究提供了理想的体内外模型.
목적 극륭구유폐화림파결전이친기성적인간암세포병건립상응적라서이식모형.방법 장형광인간암세포HCCLM3-R접충우4주령적라서간장,6주후재형광해부하관찰병획취폐급복강림파결전이조조직,경체외극륭배양,소획세포분별표기위HCCLM3-R-LM1화HCCLM3-R-LnM1.접충상술량충세포우4주령라서간장,관찰각자폐화복강림파결전이조형광수량,병여폐조직련속절편중적전이조수목진행비교.계량자료채용Wilcoxon질화검험화Kruskal-Wallis질화검험진행통계학분석.결과 HCCLM3-R-LM1、HCCLM3-R화HCCLM3-R-LnM1세포접충우라서간장후제6주,재폐화복강림파결중균능발현종류전이.3주세포접충후,라서폐화복강림파결전이조형광면적분별위8687.00±1844.63화2570.00±318.20(P＜0.001),6457.67±832.62화10 994.33±2212.31(P＜0.001),2968.67±2571.00화24 416.00±7186.13(P＜0.001),매지라서광경하폐전이조중위수분별위755、430、310개(P＜0.001),여형광정량결과상문합.결론 성공건성인간암폐화림파결친기성전이세포화라서이식모형,기중HCCLM3-R-LM1세포구유명현적폐전이특성,이HCCLM3-R-LnM1세포구유명현적림파결전이특성,위간암기관파향전이연구제공료이상적체내외모형.
Objective To establish a systematic site-specific metastatsis model of human hepatocellular carcinoma (HCC) in nude mouse.Methods HCCLM3-R cells were seeded into mice liver to establish xenograft mouse models.With the help of RFP,metastasis foci in lungs and lymph nodes in mice were detected using fluorescent stereomicroscopy and were removed.Cells derived from the metastasis foci were named HCCLM3-R-LM1 and HCCLM3-R-LnM1 respectively.HCCLM3-R-LM1 and HCCLM3-R-LnM1 cells were seeded into mice livers to analyze the lung and lymph node metastasis.Lungs of all tested mice were collected,examined by pathological evaluation and counted lung metastasis.Results Both lung and lymph node metastasis were found in HCCLM3-R-LM1,HCCLM3-R and HCCLM3-R-LnM1 cells and a significant difference was found between the lung and the lymph node metastasis levels in the three cells.The fluorescent areas (pixels) of lung and lymph node metastasis were 8687.00 ± 1844.63 versus 2570.00 ±318.20 (P = 0.0031) in HCCLM3-R-LM1 cells,6457.67±832.62 versus 10 994.33±2 212.31(P=0.0036) in HCCLM3-R cells,and 2968.67 ± 2571.00 versus 24 416.00 ± 7 186.13 (P = 0.0094) in HCCLM3-R-LnM 1 cells,respectively.The middle numbers of microscopic lung metastatic foci were 775,430 and 310in HCCLM3-R-LM1,HCCLM3-R and HCCLM3-R-LnM1 cells (P＜0.001),respectively,consist with the results quantified by RFP.Conclusion We established the systematic site-specific metastasis models which demonstrates lung- and lymph node-specific metastasis potential in nude mice and can be used as a model for researches on site-specific metastasis of HCC.