中文摘要：

目的 对来源于同一患者原发灶和跳跃转移灶的两株新建骨肉瘤细胞系-Zos和Zos-M的特性和生物学行为进行鉴定和比较研究.方法 用组织培养块法从1例骨肉瘤患者原发灶和跳跃转移灶骨肉瘤组织,分离并分别建立两株新的骨肉瘤细胞系-Zos和Zos-M;利用形态学观察、核型和细胞周期分析、生长曲线和倍增时间检测、成骨性标志检测和基质胶侵袭试验对两株细胞系进行体外鉴定.通过皮下、原位移植和实验性转移检测细胞系的体内成瘤和转移能力.应用MTT法检测Zos和Zos-M对传统化疗药物的敏感性;RT-PCR方法检测并比较二者转移相关基因的表达.结果 Zos和Zos-M体外连续培养100代后仍保持稳定,形态学检测、RT-PCR检测碱性磷酸酶、骨钙蛋白和骨桥蛋白等成骨系标志提示均符合骨肉瘤特征.Zos和Zos-M倍增时间分别为33.65 h和31.58 h.核型分析表现为非整倍体和多种染色体结构异常.Zos、Zos-M皮下和原位成瘤率均为100%,Zos-M实验性转移率为37.5%（3/8）,Zos无实验性转移（0/8）.转移相关基因表达检测表明cadherin-11在Zos-M的低表达可能与其高转移能力相关.并且Zos和Zos-M与传统骨肉瘤细胞系相比,对常用化疗药物的敏感性差.结论 人骨肉瘤细胞系Zos和Zos-M及相关动物模型的建立为治疗骨肉瘤的药物筛选提供新的模型,其具有相同的遗传背景和不同的侵袭转移能力,为骨肉瘤转移的研究提供良好的模型.

英文摘要：

Objective To characterize and compare the different biological behaviors of two novel human osteosarcoma cell lines, Zos and Zos-M, established respectively from the primary site and the skip metastasis of an osteosarcoma patient. Methods Two novel human osteosarcoma cell lines, Zos and Zos-M,were developed using tissue plant culture method. The vitro examinations included observations of morphology, analysis of karyotype and cell cycle, calculation of doubling time and growth curve, detection of osteoblastic markers and matrigel invasion assay. Subcutaneous, intratibial and intravenous inoculations into nude mice were performed to study the in vivo tumorigenicity and metastatic potentials of both cell lines.MTT were used to detect sensitivity of the cell lines to chemotherapeutic drugs. RT-PCR was performed to assess the expression of and some metastasis-related genes. Results Both cell lines proliferated actively and remained stable for more than 100 passages in vitro without interruption. The morphology and expression of osteoblastic markers of Zos and Zos-M were conformed to the characteristic of osteosarcoma. The karyotype analysis displayed aneuploidy and various structural abnormalities. The population doubling time of Zos and Zos-M were 33.65 h and 31.58 h respectively. Both cell lines were less sensitive to the current chemotherapy protocols compared to U-2OS. Zos and Zos-M were 100% tumorigenic by subcutaneous and othotopic injection. 37.5% of nude mice injected Zos-M and none of nude mice injected Zos developed lung metastasis after intravenous injection. The comparison of the expression patterns of some metastasis-related genes revealed that the decreased expression of cadherin-11 in Zos-M may correlate with a high potential of metastases. Conclusion The two novel established human osteosarcoma cell lines, Zos and Zos-M and related animal models could serve as models for the study of drug resistance and screening of new therapeutics for osteosarcoma. In addition, the study also provide tools for the s