人卵巢癌SCID小鼠移植瘤模型、细胞系建立及其生物学特性研究

目的建立人卵巢癌SCID小鼠移植瘤模型和相应体外细胞系.方法将病理证实的人卵巢浆液性乳头状腺癌手术切除标本移植于SCID小鼠皮下,成瘤后行鼠间传代,取移植瘤细胞体外分离培养、传代和建系,并应用细胞、分子生物学手段对移植瘤和建系细胞进行一系列生物学特性检测.结果历时14个月传至5代,皮下移植瘤存活率为90%,持续6个月,体外建系(OVA-319)细胞生长稳定.镜下观察组织形态学和超微结构符合原肿瘤组织基本特征;染色体分布在12～46条之间,多为异倍体,显示人类肿瘤异常染色体;流式细胞术和RT-PCR技术分析原代、体内移植瘤和OVA-319细胞结果一致,表现为瘤细胞生长活跃、细胞周期分布相仿,MAGE-2基因在mRNA水平异常表达.结论人卵巢癌SCID小鼠移植瘤模型和OVA-319细胞系为人类肿瘤的研究提供了良好的实验材料.

Biological Characteristics Studies of an Established Model of Xenografted Human Ovarian Carcinoma in SCID Mice and Its Homologous Cell Lines in Vitro

Objective To establish a model of xenografted human ovarian carcinoma in severe combined immunodificiency (SCID) mice and homologous cell lines in vitro. Method A human ovarian serous papillary carcinoma derived from a surgical specimen was transplanted in the subcutis of SCID mice. After growth they were passaged mice to mice, meanwhile a human ovarian carcinoma cell isolated from xenografted tumor in mice was cultured in vitro and coloned for six months. The biological characteristics of xenografted tumor and homologous cells were determined by cell and molecular biological techniques. Results The xenografted tumor in SCID was successively passaged for 5 generations and a successful inoculation rate (90%) was observed during a period of 14 months. A tumor cell line (OVA 319) was established in 6 months and grown steadily, which isolated from the xenografted tumor. Histology and ultrastructure of microscopy showed that xenograted tumor retained its original growth characteristics and morphological properties. The chromosomal analysis of xenografted tumor cells and OVA 319 cells revealed aneuploid pattern of 12 46 in number, exhibiting features of human carcinoma. Furthermore, the FCM analysis and RT PCR technique showed same features as those of patient with original tumor tissue and xenografted tumor and OVA 319 cells, including distribution of cell cycle and it grew rapidly, and the expression of MAGE 2 gene at the mRNA level was observed. Conclusion Establishment of xenografted human ovarian carcinoma in SCID mice and OVA 319 cell lines could be considered as a model and provided experimental material for further investigation of human ovarian tumor.