摘要
OBJECTIVE To establish a murine uterine cervical cancer cell line and to define its biological characters. METHODS Transplanted tumor tissue was used for in vitro primary culture of U14 cervical carcinoma cells. After 20 passages, we examined its morphology, chromosomes, tumorigenicity and produced a growth curve. CK was detected by immunohistochemistry, the cell cycle determined by flow cytometry and the metastatic potential assessed in 615 and C57BL/c mice. We also transfected the cells with the pEGFP-N1 plasmid. RESULTS A newly established murine cell line was passaged 50 times over a period of 10 months. The cells grow as a partially suspended culture, and are immunohistochemically CK（＋）. The cell line is characterized by a hypotetraploid karyotype, a chromosomal number of 64-68 and a doubling time of 21.8 h. Exponential growth occurs by the third and forth day of culture. Cell cycle analysis showed G1 34%, G2 26%, and 40% in the S phase. The tumorigenicity was 100% upon implantation. No mycoplasma contamination was detected. A monoclonal continuous U14-GFP cell strain which was 100% GFP （＋） was also produced. CONCLUSION We successfully established a new murine cervical U14 carcinoma cell line and an U14-GFP monoclonal strain. These cell lines are ideal for combined in vivo and in vitro tumor research.
目的:建立小鼠宫颈癌肿瘤细胞系,观察其生物学特性。方法:采用肿瘤原代培养法对小鼠U14肿瘤进行体外传代培养,定期对培养细胞进行形态学观察,进行免疫组织化学鉴定、细胞周期检查、染色体检测、细胞生长测定,进行615、C57BL/c小鼠移植观察体内成瘤情况,并用pEGFP-N1质粒转染U14细胞。结果:细胞呈贴壁和悬浮混合生长,CK阳性特征。体外连续培养10个月,传代50代,细胞倍增时间为21.83小时,细胞周期测定G1期为34%,G2期为26.4%,S期为39.6%。培养U14细胞3-4天处于对数生长期,染色体为亚四倍体核型,众数为64～84条。615小鼠、C57BL/c小鼠移植成瘤率均为100%,无支原体污染。建立了GFP(+)的U14-GFP单克隆细胞株。结论:成功建立了小鼠U14子宫颈癌细胞系及GFP标记的U14-GFP细胞株,体内移植可建立常规及带荧光标记的肿瘤模型,可用于体内体外相结合的肿瘤研究。
