维吾尔族妇女宫颈癌细胞株的建立及其生物学特性

Establishment and characterization of a new carcinoma cell line from uterine cervix of Uyghur women

目的建立维吾尔族妇女宫颈癌细胞株并观察其生物学特性，为维吾尔族妇女宫颈癌的病因学及临床治疗途径研究提供实验材料。方法无菌切除维吾尔族妇女低分化宫颈鳞状上皮细胞癌的手术标本，用酶消化法体外培养，连续传代稳定生长后，测定细胞活力，绘制细胞生长曲线。采用光镜、电镜观察细胞形态并进行细胞周期和染色体分析。观察Balb／e裸鼠接种后的成瘤情况。采用免疫荧光检测分子标志物CK17和CD44，兔超敏二步法检测分子标志物CK14、Ki-67和波形蛋白的表达情况。结果酶消化法体外培养获得维吾尔族妇女宫颈癌细胞株，持续培养12个月，传代70代，细胞呈多边形，上皮样细胞排列贴壁生长，无接触抑制，生长稳定，群体倍增时间为51．9h，瓶壁法和双层琼脂法的集落形成率分别为32．5％和15．6％。超微结构显示，细胞表面有较多突起或微绒毛，胞质内有大量杆状或棒状体和较典型的桥粒结构，核异型性明显。染色体数目在32—97条之间，众数在54～86条之间（60．3％），结构为人类染色体。裸鼠接种可形成移植瘤，组织病理形态学和患者原始肿瘤一致。细胞株分子标志物CK17、CD44、Ki-67、波形蛋白呈阳性表达，CK14呈阴性表达。结论通过酶消化法体外培养建立的维吾尔族妇女宫颈癌细胞株具有人宫颈鳞状上皮细胞癌的特点，与原发癌保持相同的生物学特性，体外连续传代培养12个月，细胞形态及生长周期恒定，有望成为一个稳定的细胞株。

Objective To establish a uterine cervical carcinoma cell line of Uyghur ethnical background and to evaluate the related biological characteristics for future biomedical investigations of diseases in the Uyghur population. Methods Poorly-differentiated squamous cell carcinoma specimens of Uyghur patients were obtained and cultured in vitro by enzymatic digestion method, followed by continuous passaging to reach a stable growth determined by cell viability and growth curve. Morphological study, cell cycling and chromosomal analysis were performed. Tumorigenesis study was conducted by inoculation of nude mice. Biomarker（ CK17, CD44, Ki-67, CK14 and vimentin） expression was detected by immunofluorescence and immunocytochemical techniques. Results A cervical carcinoma cell line was successfully established and maintained for 12 months through 70 passages. The cell line had a stable growth with a population doubling time of 51.9 h. Flask method and double agar-agar assay showed that the cell line had colony- forming rates of 32. 5% and 15.6%, respectively. Uhrastructural evaluation demonstrated numerous cell surface protrusions or microvilli, a large number of rod-shape structures in cytoplasm, typical desmosomes and nuclear atypia. Chromosomal analysis revealed human karyotype with the number of chromosomes per cell varying from 32-97 with a majority of 54-86 （60. 3% ）. Xenogeneic tumors formed in nude mice showed histological structures identical to those of the primary tumor. The ceils had high expression of CK17, CD44, Ki-67 and vimentin but no CK14 expression. Conclusions A cervical carcinoma cell line from a female Uyghur patient is successfully established. The cell line has the characteristics of human cervical squamous cell carcinoma, and it is stable with maintaining the characteristic biological and morphological features invitro for more than 12 months, therefore, qualified as a stable cell line for further biomedical research.