转染﹀G基因HHCC细胞在裸鼠体内的转移特性

Metastatic character of β-Glucuronidase-cDNA transfected HHCC in nude mice

目的 利用转染 βG基因的 HHCC细胞构建裸鼠转移模型 .方法 将 HHCC细胞和转染 βG基因 HHCC细胞体外培养 ,观察细胞形态结构 ,流式细胞仪测定细胞周期 ,并采用裸鼠脾脏种植法对转βG基因 HHCC细胞的转移特性进行研究 .接种后 6 0 d,收集转移灶 ,常规制备光镜片 ,HE染色 .结果 转染βG基因 HHCC细胞内多见颗粒性物质 ,微绒毛及突起增多 ,溶酶体增多 ,内质网及核糖体丰富 ,糖元颗粒堆积 ,某些细胞出现变性、坏死 .转 βG基因 HHCC和 HHCC相比较 ,其细胞周期发生明显改变 ,S期细胞增多 (P<0 .0 5 ) ,G2期细胞减少 (P<0 .0 5 ) ;裸鼠脾脏种植后 6 0 d,发现实验组出现广泛转移在肝、肾、肠系膜淋巴结的肉眼可见的转移灶 ,而对照组仅出现肝内播散 .结论 转染βG基因可增强

AIM To establish metastasis model in nude mice by inoculationβ Glucuronidase cDNA transfected HHCC. METHODS The βG cDNA transfected human hepatic cell line(HHCC)was cultured in vitro . Two types of cells were analyzed on its morphology and cell cycling. The cell metastasis properties were studied by spleen inoculation in nude mice. Sixty days later, the metastasis focus was fixed and stained HE by traditional method. RESULTS Granular substances could be observed in βG cDNA transfected HHCC. The morphologic changes of HHCC βG, such as ① microvillus and process increasing,②cell degeneration and necrosis,③increased endoplasmic reticulum and ribosome and ④the marked increasing of lsosomes could be observed under the transmission electron microscope. Compared with HHCC βG, the transfected HHCC cell cycling changed distinctly. The cells in S phase increased( P <0.05) while those in G2 phase decreased ( P <0.05). After spleen inoculation in nude mice, the experiment group displayed widely the metastasis character. The metastatic focus could be seen in liver, renal and mesenteric lymphnodes. The control group could only form inside liver invading and disseminating. CONCLUSION TransfectingβG cDNA can enhance HHCC metastasis potential in nude mice.